Colibri System

K193138 · Copan Wasp Srl · QQV · Dec 27, 2021 · Microbiology

Device Facts

• Record ID: K193138
• Device Name: Colibri System
• Applicant: Copan Wasp Srl
• Product Code: QQV · Microbiology
• Decision Date: Dec 27, 2021
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3378
• Device Class: Class 2

Indications for Use

The Colibrí System is an in vitro diagnostic device comprised of the Colibrí Vision System and Colibrí Preparation Station for use with the bioMérieux VITEK MS or Bruker MALDI Biotyper CA mass spectrometry systems for qualitative identification of isolated colonies of Gram-negative and Gram-positive bacterial species grown on solid culture media. The Colibrí System is a semiautomated pre-analytical processor that picks isolated colonies designated by the operator and uses a pipetting system to prepare MALDI-TOF MS (Matrix-Assisted Laser Desorption/Ionization-Time Of Flight Mass Spectrometry) target slides. The Colibrí software records the identity of each sample and its position on the target slide and communicates this information electronically to the MALDI-TOF MS analyzer. The Colibrí System is intended for use by trained healthcare professionals in clinical laboratories in conjunction with other clinical and laboratory findings, including Gram staining, to aid in the diagnosis of bacterial infections. The Colibrí System has not been validated for use in identification of yeast species.

Device Story

Semi-automated pre-analytical processor for MALDI-TOF MS target preparation. Input: culture plates with isolated bacterial colonies. Process: operator uses Colibrí Vision System to acquire digital images and designate colonies; Colibrí Preparation Station automatically picks designated colonies, spots them onto MALDI targets, and overlays with matrix. Output: prepared MALDI target slides; electronic communication of sample identity and position to downstream mass spectrometer (bioMérieux VITEK MS or Bruker MALDI Biotyper CA). Used in clinical laboratories by trained professionals. Benefits: automates manual colony picking and spotting, reducing variability and labor; improves workflow efficiency. Healthcare providers use resulting organism identification to aid in diagnosis of bacterial infections.

Clinical Evidence

Bench testing only. Studies included colony picking accuracy (100% correct picking), positional effect, inclusivity (123-124 strains), specificity (off-panel strains), reproducibility (1800 spots), cross-contamination, and colony/spot stability. Overall agreement for VITEK MS was 97.2% (inclusivity) and 99.9% (reproducibility). For Bruker MALDI Biotyper CA, performance varied by species/target; Gram-positive identification showed lower confidence, with labeling recommending manual eDT/Ext procedures for low-confidence results. No false positives or wrong identifications reported in key studies.

Technological Characteristics

System comprises Colibrí Vision System (digital imaging) and Colibrí Preparation Station (automated pipetting). Uses direct transfer method for MALDI-TOF MS target preparation. Compatible with VITEK MS-DS, MBT Biotarget 96, and 48-spot reusable targets. Connectivity: electronic data transmission to mass spectrometer. Software: proprietary Colibrí Vision and Preparation software. Sterilization: not applicable (instrument).

Indications for Use

Indicated for qualitative identification of isolated Gram-negative and Gram-positive bacterial colonies grown on solid culture media. Intended for use by trained healthcare professionals in clinical labs to aid in bacterial infection diagnosis. Not validated for yeast, mold, Nocardia, or Mycobacteria.

Regulatory Classification

Identification

A clinical mass spectrometry microorganism identification and differentiation system is a qualitative in vitro diagnostic device intended for the identification and differentiation of microorganisms from processed human specimens. The system acquires, processes, and analyzes spectra to generate data specific to a microorganism(s). The device is indicated for use in conjunction with other clinical and laboratory findings to aid in the diagnosis of bacterial and fungal infection.

Special Controls

Clinical mass spectrometry microorganism identification and differentiation system must comply with the following special controls:

*Classification.* Class II (special controls). The special controls for this device are:(1) The intended use statement must include a detailed description of what the device detects, the type of results provided to the user, the clinical indications appropriate for test use, and the specific population(s) for which the device is intended, when applicable. (2) Any sample collection device used must be FDA-cleared, -approved, or -classified as 510(k) exempt with an indication for in vitro diagnostic use. (3) The labeling required under § 809.10(b) of this chapter must include: (i) A detailed device description, including all device components, control elements incorporated into the test procedure, instrument requirements, ancillary reagents required but not provided, and a detailed explanation of the methodology and all pre-analytical methods for processing of specimens, and algorithm used to generate a final result. This must include a description of validated inactivation procedure(s) that are confirmed through a viability testing protocol, as applicable. (ii) Performance characteristics for all claimed sample types from clinical studies with clinical specimens that include prospective samples and/or, if appropriate, characterized samples. (iii) Performance characteristics of the device for all claimed sample types based on analytical studies, including limit of detection, inclusivity, reproducibility, interference, cross-reactivity, interfering substances, carryover/cross-contamination, sample stability, and additional studies regarding processed specimen type and intended use claims, as applicable. (iv) A detailed explanation of the interpretation of test results for clinical specimens and acceptance criteria for any quality control testing. (4) The device's labeling must include a prominent hyperlink to the manufacturer's website where the manufacturer must make available their most recent version of the device's labeling required under § 809.10(b) of this chapter, which must reflect any changes in the performance characteristics of the device. FDA must have unrestricted access to this website, or manufacturers must provide this information to FDA through an alternative method that is considered and determined by FDA to be acceptable and appropriate. (5) Design verification and validation must include: (i) Any clinical studies must be performed with samples representative of the intended use population and compare the device performance to results obtained from an FDA-accepted reference method and/or FDA-accepted comparator method, as appropriate. Documentation from the clinical studies must include the clinical study protocol (including predefined statistical analysis plan, if applicable), clinical study report, and results of all statistical analyses. (ii) Performance characteristics for analytical and clinical studies for specific identification processes for the following, as appropriate: (A) Bacteria, (B) Yeasts, (C) Molds, (D) Mycobacteria, (E) Nocardia, (F) Direct sample testing ( *e.g.,* blood culture),(G) Antibiotic resistance markers, and (H) Select agents ( *e.g.,* pathogens of high consequence).(iii) Documentation that the manufacturer's risk mitigation strategy ensures that their device does not prevent any device(s) with which it is indicated for use, including incorporated device(s), from achieving their intended use ( *e.g.,* safety and effectiveness of the functions of the indicated device(s) remain unaffected).(iv) A detailed device description, including the following: (A) Overall device design, including all device components and all control elements incorporated into the testing procedure. (B) Algorithm used to generate a final result from raw data ( *e.g.,* how raw signals are converted into a reported result).(C) A detailed description of device software, including validation activities and outcomes. (D) Acquisition parameters ( *e.g.,* mass range, laser power, laser profile and number of laser shots per profile, raster scan, signal-to-noise threshold) used to generate data specific to a microorganism.(E) Implementation methodology, construction parameters, and quality assurance protocols, including the standard operating protocol for generation of reference entries for the device. (F) For each claimed microorganism characteristic, a minimum of five reference entries for each organism (including the type strain for microorganism identification), or, if there are fewer reference entries, a clinical and/or technical justification, determined by FDA to be acceptable and appropriate, for why five reference entries are not needed. (G) DNA sequence analysis characterizing all type strains and at least 20 percent of the non-type strains of a species detected by the device, or, if there are fewer strain sequences, then a clinical and/or technical justification, determined by FDA to be acceptable and appropriate, must be provided for the reduced number of strains sequenced. (H) As part of the risk management activities, an appropriate end user device training program, which must be offered as an effort to mitigate the risk of failure from user error.

Predicate Devices

• bioMérieux VITEK MS System (K181412)
• Bruker MALDI Biotyper CA System (DEN170081)

Submission Summary (Full Text)

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FDA

U.S. FOOD & DRUG

ADMINISTRATION

# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

INSTRUMENT ONLY

## I Background Information:

A 510(k) Number

K193138

B Applicant

COPAN WASP S.r.l.

C Proprietary and Established Names

Colibrí System

D Regulatory Information

| Product Code(s) | Classification | Regulation Section | Panel |
| --- | --- | --- | --- |
| QQV | Class II | 21 CFR 866.3378 - Clinical mass spectrometry microorganism identification and differentiation system | MI - Microbiology |
| QBN | Class II | 21 CFR 866.3378 - Clinical mass spectrometry microorganism identification and differentiation system | MI - Microbiology |

## II Submission/Device Overview:

A Purpose for Submission:

To obtain a substantial equivalence determination for the Copan Colibrí System.

B Type of Test:

Qualitative in vitro diagnostic device for identification and differentiation of microorganisms

Food and Drug Administration

10903 New Hampshire Avenue

Silver Spring, MD 20993-0002

www.fda.gov

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cultured from human specimens by automation of target preparation for mass spectrometry analysis.

## III Intended Use/Indications for Use:

### A Intended Use(s):
See Indications for Use below.

### B Indication(s) for Use:
The Colibrí System is an in vitro diagnostic device comprised of the Colibri Vision System and Colibri Preparation System for use with the bioMérieux VITEK MS or Bruker MALDI Biotyper CA mass spectrometry systems for the qualitative identification of isolated colonies of Gram-negative and Gram-positive bacterial species grown on solid culture media. The Colibrí System is a semi-automated pre-analytical processor that picks isolated colonies designated by the operator and uses a pipetting system to prepare MALDI-TOF MS (Matrix-Assisted Laser Desorption/Ionization-Time Of Flight Mass Spectrometry) target slides. The Colibrí software records the identity of each sample and its position on the target slide and communicates this information electronically to the MALDI-TOF MS analyzer.

The Colibrí System is intended for use by trained healthcare professionals in clinical laboratories in conjunction with other clinical and laboratory findings, including Gram staining, to aid in the diagnosis of bacterial infections.

The Colibrí System has not been validated for use in identification of yeast species.

### C Special Conditions for Use Statement(s):
Rx - For Prescription Use Only

IVD - For In Vitro Diagnostic Use Only.

Special Instruments for Use:
bioMérieux VITEK MS
Bruker MALDI Biotyper for Clinical Applications (MBT-CA)
When using the Copan Colibrí System, refer to the most recent version of the bioMérieux VITEK MS or Bruker MALDI Biotyper CA system labeling.

When the Copan Colibrí System is used with the bioMérieux VITEK MS, use only whole or bi-plates of the culture media listed below:
- Columbia Agar + 5% sheep blood (bioMérieux)
- Trypticase Soy Agar + 5% sheep blood (Becton Dickinson)
- MacConkey Agar (bioMérieux)
- Chocolate Agar (Becton Dickinson)

When the Copan Colibrí System is used with the Bruker MALDI Biotyper CA, use only whole or bi-plates of the culture media listed below:
- Columbia Agar + 5% sheep blood (bioMérieux)
- Trypticase Soy Agar + 5% sheep blood (Becton Dickinson)

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- MacConkey Agar (bioMérieux)
- Chocolate Agar (Becton Dickinson)
- Columbia Agar + 5% sheep blood supplemented with colistin and nalidixic acid (CNA) (Becton Dickinson)
- Bordet Gengou Agar + 15% sheep blood (Becton Dickinson)

Use of the Copan Colibrí System to prepare samples for analysis with the bioMérieux VITEK MS and Bruker MALDI Biotyper CA System was evaluated using only the species listed in the Inclusivity Study in the Performance Characteristics section.

The Colibrí System is not intended for and has not been validated for use in the identification of yeasts, molds, Nocardia or Mycobacteria.

Colibrí System has been validated for the direct spotting of isolated bacterial colony(ies) grown on solid media and overlaid with matrix. Other methods of target preparation such as overlay with formic acid or extraction have not been validated and should be conducted manually per the applicable MALDI-TOF MS analyzer instructions for use.

Use of the Copan Colibrí System to prepare targets from cultures of Gram-positive organisms resulted in a lower proportion of High Confidence Log(scores) with the Bruker MALDI Biotyper CA System. The operator is instructed to repeat testing of the isolate manually using the extended Direct Transfer (eDT) or Extraction (Ext) Sample Preparation Procedure if a low-confidence identification or no identification result is obtained.

## IV Device/System Characteristics:

### A Device Description:

The Copan Colibrí System automates various steps in the preparation of targets for the bioMérieux VITEK MS and Bruker MALDI Biotyper CA systems that use Matrix-Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) for the identification of organisms grown on plated culture media. The Colibrí System comprises the Colibrí Vision System for digital imaging of culture plates and operator-designation of colonies for picking and the Colibrí Preparation Station for automated picking of designated colonies and spotting of the MALDI targets.

The Colibrí System follows the direct transfer method of sample deposition on the target slides for both the VITEK MS and MALDI Biotyper CA in which colonies are spotted directly on targets and then overlaid with MALDI matrix. To use the system, the operator manually sorts culture plates based on the type of medium, presence of visible growth and Gram stain results, and applies a unique barcode to each plate for sample identification. The operator then loads the pre-sorted plates into the Colibrí Vision System for digital image acquisition and designates well-isolated bacterial colonies for identification using the Image Reading Interface.

Following image acquisition and colony designation, the operator manually transfers the appropriate plates to the Colibrí Preparation Station for automated colony picking, spotting of the

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target and overlay of the dried target spots with matrix. The coordinates of the individual colonies for picking are retrieved automatically by the Colibrí Preparation Station from the Colibrí Vision System.

Once the Colibrí System has finished processing the culture plates, the operator removes the target slides and manually spots the appropriate controls for the applicable downstream MALDI-TOF MS instrument before initiating analysis of the samples. The location of each sample to be identified is sent electronically from the Colibrí System to the applicable downstream MALDI-TOF MS instrument prior to processing of the target. Identification results are reported by the MALDI-TOF MS instruments according to the algorithms specific to each system.

B Instrument Description Information:

| Modes of Operation | Yes | No |
| --- | --- | --- |
| Does the applicant’s device contain the ability to transmit data to a computer, webserver, or mobile device? | ☑ | ☐ |
| Does the applicant’s device transmit data to a computer, webserver, or mobile device using wireless transmission? | ☐ | ☑ |
| Software | | |
| FDA has reviewed applicant’s Hazard Analysis and software development processes for this line of product types. | ☑ | ☐ |

1. Instrument Name:

Copan Colibrí System comprised of the Colibrí Vision System and Colibrí Preparation Station

2. Specimen Sampling and Handling:

Culture plates for processing by the Colibrí System are sorted manually to identify those with isolated colonies of Gram-positive or Gram-negative bacteria and then labeled by applying a linear barcode to the base of the plate. After loading the plates into the Colibrí Vision System, the operator must select the appropriate plate type (i.e., culture medium type) for image acquisition to ensure that the appropriate optical parameters are applied. Following image acquisition, the operator designates well-isolated colonies for picking and then manually transfers the plates from the Colibrí Vision System to the Colibrí Preparation Station, where the designated colonies are picked automatically and used to prepare the appropriate MALDI target, using either the bioMérieux VITEK MS-DS target slides or the Bruker US IVD 48-spot reusable or 96-spot disposable targets.

3. Specimen Identification:

Culture plates for processing are identified by the Colibrí Vision System by scanning the manually applied linear barcode on the side of each plate. The barcode is used to orientate the plate and, together with the plate's geometric center, also used to define the Cartesian coordinates of each of the colonies that are designated for picking. This information is retrieved by the Colibrí Preparation Station, which picks the designated colonies from the

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agar surface and deposits them into target spots and overlays the spots with the appropriate matrix (bioMérieux VITEK MS-CHCA matrix or the Bruker US IVD HCCA portioned) using the pipetting unit. The Colibrí Preparation Station records the location on the MALDI target into which each designated colony is spotted. The layout of the target is sent electronically to the downstream MALDI-TOF MS instrument prior to analysis. The bioMérieux VITEK MS or the Bruker MALDI Biotyper CA system then performs species identification according to their respective procedures. There are no changes to the methods of data analysis or result algorithms for either the VITEK MS or MALDI Biotyper CA compared to those used in the 510(k)-cleared devices.

## 4. Calibration:

Three calibrations are performed for the Copan Colibrí System.

### Set-up Calibration

Set-up calibration refers to the camera and positional calibrations that are performed by the technical engineer as part of the camera and vision system hardware setup during the initial device installation.

### Auto-calibration

Auto-calibration is applicable to the Colibrí Preparation System and is performed automatically by the instrument following mechanical calibration by the technical engineer during initial device installation and periodically thereafter during preventative maintenance (every 6 months) over the entire device lifecycle. During this procedure, the Colibrí Vision Software performs specific calls to the cameras and to the hardware related to the visual system to check that all the mechanical references can be found inside the positioning tolerances, that the input/output interfaces are responsive and that the spaces subject to verification during the cycle are clean and free from obstacles.

### Run-time Calibration Checks

This calibration refers to all activities that the Image Acquisition and Colibrí Vision software perform during normal usage.

## 5. Quality Control:

Quality Control testing is performed according to manufacturer's instructions of the bioMérieux VITEK MS and Bruker MALDI Biotyper mass spectrometry systems using cultures of Escherichia coli ATCC 8739 (calibrator strain) and Klebsiella aerogenes ATCC 13048 (positive control strain) grown on Columbia or Trypticase Soy Agar plates (both with 5% sheep blood) for the VITEK MS or US IVD Bacterial Test Standard (BTS) containing lyophilized Escherichia coli ATCC 25922 reconstituted in Standard Solvent for the MALDI Biotyper CA. Spots are prepared using the appropriate mode of target deposition (single or duplicate) and in each case the expected MALDI-TOF MS identification result(s) should be obtained with a high confidence value (VITEK MS) or Log(Score) (MALDI Biotyper CA). Negative quality control spots are prepared using matrix alone. Quality control testing was performed during analytical performance testing, and all results were obtained as expected. Results are summarized below.

| Quality Control | N | # agreement (%) |
| --- | --- | --- |
| Klebsiella aerogenes ATCC 13048 | 207 | 207 (100) |
| US IVD Bacterial Test Standard | 234 | 234 (100) |

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Negative control (matrix alone) 441 441 (100)
TOTAL 882 882 (100)

In addition, culture plates of *E. coli* (ATCC 25922) grown on Columbia or Trypticase Soy Agar plates (both with 5% sheep blood) are processed through the Copan Colibri System workflow as described in the Colibrí Vision System and Colibrí Preparation Station Operator Manuals. *E. coli* ATCC 25922 was evaluated as a test strain in all analytical studies, excluding the Specificity Study and all results were obtained as expected.

V Substantial Equivalence Information:

A Predicate Device Name(s): bioMérieux VITEK MS System

B Predicate 510(k) Number(s): K181412

C Comparison with Predicate(s):

| Device & Predicate Device(s): | K193138 | K181412 | DEN170081 |
| --- | --- | --- | --- |
| Device Trade Name | Copan Colibrí System | bioMérieux VITEK MS | Bruker MALDI Biotyper CA System |
| Intended Use/Indications for Use | The Colibrí System is an in vitro diagnostic device comprised of the Colibrí Vision System and Colibrí Preparation Station for use with the bioMérieux VITEK MS or Bruker MALDI Biotyper CA mass spectrometry systems for qualitative identification of isolated colonies of Gram-negative and Gram-positive bacterial species grown on solid culture media. The Colibrí System is a semi-automated pre-analytical processor that picks isolated colonies designated by the operator and uses a pipetting system to prepare MALDI-TOF MS (Matrix-Assisted Laser Desorption/Ionization-Time Of Flight Mass Spectrometry) target slides. The Colibrí software records the identity of each sample and its position on the target slide and communicates this information electronically to the MALDI-TOF MS | VITEK MS is a mass spectrometry system using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) for the identification of microorganisms cultured from human specimens. The VITEK MS system is a qualitative in vitro diagnostic device indicated for use in conjunction with other clinical and laboratory findings to aid in the diagnosis of bacterial, yeast and mould infections. (list of claimed organisms omitted for brevity; refer to K181412) | The MALDI Biotyper CA System is a mass spectrometer system using matrix-assisted laser desorption/ ionization - time of flight (MALDI-TOF) for the identification and differentiation of microorganisms cultured from human specimens. The MALDI Biotyper CA System is a qualitative in vitro diagnostic device indicated for use in conjunction with other clinical and laboratory findings to aid in the diagnosis of bacterial and fungal infections. (list of validated organisms omitted for brevity; refer to DEN170081) |

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| Device & Predicate Device(s): | K193138 | K181412 | DEN170081 |
| --- | --- | --- | --- |
| Device Trade Name | Copan Colibrí System | bioMérieux VITEK MS | Bruker MALDI Biotyper CA System |
| | analyzer.
The Colibrí System is intended for use by trained healthcare professionals in clinical laboratories in conjunction with other clinical and laboratory findings, including Gram staining, to aid in the diagnosis of bacterial infections.
The Colibrí System has not been validated for use in identification of yeast species. | | |
| General Device Characteristic Similarities | | | |
| Sample Type | Isolated colonies on plated culture media
Acceptable media when used with the VITEK MS:
• Columbia blood agar with 5% sheep blood
• Trypticase soy agar with 5% sheep blood
• Chocolate agar
• MacConkey agar

Acceptable media when used with the MALDI Biotyper CA:
• Columbia blood agar with 5% sheep blood
• Trypticase soy agar with 5% sheep blood
• Chocolate agar
• MacConkey agar
• Columbia CNA agar with 5% sheep blood
• Bordet Gengou Agar with 15% sheep blood | Isolated colonies on plated culture media
Acceptable media:
• Columbia blood agar with 5% sheep blood
• Trypticase soy agar with 5% sheep blood
• Chocolate agar
• MacConkey agar | Isolated colonies on plated culture media
Acceptable media:
• Columbia blood agar with 5% sheep blood
• Trypticase soy agar with 5% sheep Blood
• Chocolate agar
• MacConkey Agar
• Columbia CNA agar with 5% sheep blood
• Bordet Gengou Agar with 15% sheep blood |
| Target Preparation | Direct Transfer of colony to target spot and overlay with matrix | Same | Same |
| Quality Control Preparation | Manual | Same | Same |
| Calibration/Quality Controls | Escherichia coli ATCC 8739 and Klebsiella | Escherichia coli ATCC 8739 (calibrator strain) | US IVD Bacterial Test Standard (BTS) |

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| Device & Predicate Device(s): | K193138 | K181412 | DEN170081 |
| --- | --- | --- | --- |
| Device Trade Name | Copan Colibrí System | bioMérieux VITEK MS | Bruker MALDI Biotyper CA System |
| | aerogenes ATCC 13048 -OR- US IVD Bacterial Test Standard (BTS) | and Klebsiella aerogenes ATCC 13048 (positive control strain) cultured on plated media | lyophilized Escherichia coli ATCC 25922 reconstituted in Standard Solvent |
| Matrix | VITEK MS-CHCA -OR- US IVD HCCA Portioned | VITEK MS-CHCA | US IVD HCCA Portioned |
| Targets | VITEK MS-DS target slide -OR- MBT Biotarget 96 US IVD (96-spot disposable) target US IVD 48 Spot (48-spot reusable) target | VITEK MS-DS target slide | MBT Biotarget 96 US IVD (96-spot disposable) target US IVD 48 Spot (48-spot reusable) target |
| Colonies per Target Spot | One | Same | Same |
| MALDI-TOF MS Analyzer | bioMérieux VITEK MS -OR- Bruker MALDI Biotyper CA | bioMérieux VITEK MS | Bruker MALDI Biotyper CA |
| Results Reported | Organism identification | Same | Same |
| General Device Characteristic Differences | | | |
| Target Organisms | Gram-positive/Gram-negative bacteria only | Gram-positive/Gram-negative bacteria, yeasts, Mycobacteria, Nocardia and molds | Gram-positive/Gram-negative bacteria and yeasts |
| Colony/Plate Visualization | Digital image from the Colibrí Vision System module | Direct | Direct |
| Target Preparation | Automated | Manual | Manual |
| Alternative Methods of Target Preparation | None | None | Extended Direct Transfer (eDT) Test Procedure Extraction (Ext) Test Procedure |
| Compatible Culture Media | VITEK MS: • Columbia Agar + 5% sheep blood • Trypticase Soy Agar + 5% sheep blood • MacConkey Agar | Same PLUS: • BacT/ALERT MP • Brucella agar base • Buffered charcoal yeast extract • Campylosel agar | Same PLUS: • Brucella Agar + 5% horse blood • CDC Anaerobe Agar + 5% sheep blood • CDC Anaerobe Agar + 5% |

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| Device & Predicate Device(s): | K193138 | K181412 | DEN170081 |
| --- | --- | --- | --- |
| Device Trade Name | Copan Colibri System | bioMérieux VITEK MS | Bruker MALDI Biotyper CA System |
| | • Chocolate Agar
Bruker MALDI Biotyper CA: as above PLUS
• Columbia Agar + 5% sheep blood supplemented with colistin and nalidixic acid (CNA)
• Bordet Gengou Agar + 15% sheep blood | • chromID CPS
• Coletsos
• Lowenstein-Jensen
• MGIT
• Middlebrook 7H10 agar
• Middlebrook 7H11 agar
• Modified Sabouraud dextrose agar (glucose: 20 g/L - pH: 6.1)
• Potato dextrose agar
• Sabouraud dextrose agar (glucose: 40 g/L pH: 5.6)
• Sabouraud dextrose agar with Gentamicin & Chloramphenicol
• Trypticase soy agar
• Trypticase soy agar with neutralizers | sheep blood with phenylethyl alcohol
• CDC Anaerobe Laked Sheep Blood Agar with kanamycin and vancomycin
• Bacteroides Bile Esculin Agar with Amikacin
• Clostridium difficile Agar + 7% sheep blood
• Sabouraud-Dextrose Agar
• Campylobacter Agar with 5 Antimicrobics and 10% Sheep Blood
• Buffer Charcoal Yeast Extract Agar
• Buffered Charcoal Yeast Extract Selective Agar with polymyxin, anisomycin and vancomycin
• Modified Thayer-Martin Agar |
| Technology | Automated imaging and pipetting system with downstream MALDI-TOF MS organism identification | Direct visual inspection and selection and manual preparation of MALDI-TOF MS targets for organism identification | Direct visual inspection and selection and manual preparation of MALDI-TOF MS targets for organism identification |

VI Standards/Guidance Documents Referenced:

IEC 61010-1. Safety Requirements for Electrical Equipment for Measurement, Control and Laboratory Use - Part 1: General Requirements [including corrigendum 1] (Third Edition; 2010-06).

IEC 61010-2-010. Safety requirements for electrical equipment for measurement, control and laboratory use - Part 2-010: Particular requirements for laboratory equipment for the heating of materials.

IEC 61010-2-081. Safety requirements for electrical equipment for measurement, control and laboratory use - Part 2-081: Particular requirements for automatic and semi-automatic laboratory equipment for analysis and other purposes.

IEC 61010-2-101. Safety requirements for electrical equipment for measurement, control and laboratory use - Part 2-101: Particular requirements for in vitro diagnostic (IVD) medical equipment.

IEC 61326-1. Electrical equipment for measurement, control and laboratory use - EMC

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requirements - Part 1: General requirements.

IEC 61326-2-6: Electrical equipment for measurement, control and laboratory use - EMC requirements - Part 2-6: Particular requirements - In vitro diagnostic (IVD) medical equipment.

IEC 62304. Medical Device Software - Software Life Cycle Processes (Edition 1.1; 2015-06).

IEC 60825-1. Safety of Laser Products - Part 1: Equipment Classification and Requirements [Including: Technical Corrigendum 1 (2008), Interpretation Sheet 1 (2007), Interpretation Sheet 2 (2007)] (Second Edition; 2007-03).

IEC 62366-1. Medical Devices - Part 1: Application of Usability Engineering to Medical Devices [including Corrigendum 1 (2016)] (First Edition; 2015-02).

ISO 15223-1. Medical Devices - Symbols to be Used with Medical Device Labels, Labelling and Information to be Supplied - Part 1: General Requirements (Third Edition; 2016-11-01).

ISO 14971. Medical Devices - Application of Risk Management to Medical Devices (Second Edition; 2007-03-01).

CLSI. Methods for the Identification of Cultured Microorganisms Using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry. $1^{\mathrm{st}}$ ed. CLSI guideline M58. Wayne, PA: Clinical and Laboratory Standards Institute; 2017.

# VII Performance Characteristics (if/when applicable):

The performance of the Copan Colibri System was evaluated in conjunction with the bioMérieux VITEK MS and Bruker MALDI Biotyper CA instruments using the confidence level and interpretive criteria for species identification as described in the labeling for each respective mass spectrometer, as summarized in Tables 1 and 2.

Table 1. Interpretive criteria for species identification using the bioMérieux VITEK MS.

| Confidence Level | Number of Organisms Reported | Confidence Value |
| --- | --- | --- |
| Good | 1 | 60 - 99.9% |
| Low Discrimination | 2 - 41 | Sum = 100% |
| No identification | None or > 41 | Not applicable or sum < 100% |

1 Additional testing required to determine or confirm organism identity.

Table 2. Interpretive criteria for species identification using the Bruker MALDI Biotyper CA.

| Confidence Level | Log(Score) Value |
| --- | --- |
| High | ≥ 2.00 |
| Low | 1.70 - 1.991 |
| No identification | < 1.701 |

1 Additional testing required to determine or confirm organism identity.

# A Analytical Performance:

# 1. Precision/Reproducibility:

The reproducibility of bacterial identification results obtained following automated sample preparation by the Colibri System was evaluated in separate studies with the VITEK MS and MALDI Biotyper CA analyzers (both with 48-spot targets). In each study, three Colibri Systems (each comprised of a Colibri Vison System and Colibri Preparation Station) were

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used to prepare targets from overnight (18 to 24 hour) cultures of representative, clinically relevant Gram-positive and Gram-negative bacterial species grown on Trypticase Soy Agar with $5\%$ sheep blood. The three Colibri Systems were used in rotation in conjunction with a single mass spectrometry instrument of each type. Each Colibri System was used to prepare targets of each bacterial species on five separate days. On each day on which testing was performed, two operators with different levels of microbiological experience each designated two colonies for automated picking from digital images of each of three culture plates corresponding to each bacterial species (3 Colibri Systems $\times 5$ days $\times 2$ operators $\times 3$ plates $\times 2$ colonies $= 180$ colonies per species). Following designation of the colonies for picking using the Colibri Vision System, the culture plates were transferred by hand to the corresponding Colibri Preparation Station. After automated processing of the designated colonies, the appropriate calibrators were spotted manually onto the targets, which were then transferred to the applicable downstream MALDI-TOF MS analyzer. The reported organism identity was compared to the expected identity, i.e. the known species identifications for the reference isolates, and within-run, between-run and between-user reproducibility were investigated. Reproducibility of Copan Colibri with the bioMérieux VITEK MS and with the Bruker MALDI Biotyper CA are outlined below in sections $a$ and $b$ , respectively.

# a. Reproducibility of Copan Colibri with the bioMérieux VITEK MS

Overall, for the Copan Colibri System in conjunction with the bioMérieux VITEK MS, there was $89.9\%$ agreement (1619/1800) between the reported Good Confidence identification results and the expected identity of each colony in the Reproducibility Study (for all Gram-positive and Gram-negative species combined) (Table 3). In addition, all 180 colonies of Enterobacter cloacae were reported with Low Discrimination as Enterobacter cloacae/Enterobacter asburiae, in accordance with the labeling for the VITEK MS analyzer. Therefore, in total 1799/1800 $(99.9\%)$ results obtained were as expected. No significant differences in agreement were observed using Fisher's Exact test between instruments and operators, and none of the samples in the study were incorrectly identified. The reproducibility of the Copan Colibri System with VITEK MS was therefore determined to be acceptable.

Table 3. Reproducibility of the Copan Colibri System with the bioMérieux VITEK MS.

| Gram Stain Phenotype | Species/Strain | Colibri System | VITEK MS Identification Result (%) | | |
| --- | --- | --- | --- | --- | --- |
| | | | Good Confidence | Low Discrimination | None |
| Gram-Negative | Enterobacter cloacae1ATCC 13047 | 008 | 0 (0) | 60 (100) | 0 (0) |
| | | 009 | 0 (0) | 60 (100) | 0 (0) |
| | | 010 | 0 (0) | 60 (100) | 0 (0) |
| | | All | 0 (0) | 180 (100) | 0 (0) |
| | Escherichia coliATCC 25922 | 008 | 60 (100) | 0 (0) | 0 (0) |
| | | 009 | 60 (100) | 0 (0) | 0 (0) |
| | | 010 | 60 (100) | 0 (0) | 0 (0) |
| | | All | 180 (100) | 0 (0) | 0 (0) |
| | Klebsiella pneumoniaeATCC BAA 1705 | 008 | 60 (100) | 0 (0) | 0 (0) |
| | | 009 | 60 (100) | 0 (0) | 0 (0) |
| | | 010 | 60 (100) | 0 (0) | 0 (0) |
| | | All | 180 (100) | 0 (0) | 0 (0) |
| | Proteus mirabilisATCC 7002 | 008 | 60 (100) | 0 (0) | 0 (0) |
| | | 009 | 60 (100) | 0 (0) | 0 (0) |
| | | 010 | 60 (100) | 0 (0) | 0 (0) |
| | | All | 180 (100) | 0 (0) | 0 (0) |
| | Pseudomonas aeruginosaATCC 27853 | 008 | 60 (100) | 0 (0) | 0 (0) |
| | | 009 | 60 (100) | 0 (0) | 0 (0) |
| 010 | | 60 (100) | 0 (0) | 0 (0) | |

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| Gram Stain Phenotype | Species/Strain | Colibri System | VITEK MS Identification Result (%) | | |
| --- | --- | --- | --- | --- | --- |
| | | | Good Confidence | Low Discrimination | None |
| | | All | 180 (100) | 0 (0) | 0 (0) |
| | All Gram-Negative species (n=900) | 008 | 240 (100) | 60 (20.0) | 0 (0) |
| | | 009 | 240 (100) | 60 (20.0) | 0 (0) |
| | | 010 | 240 (100) | 60 (20.0) | 0 (0) |
| | | All | 720 (80.0) | 180 (20.0) | 0 (0) |
| Gram-Positive | Enterococcus faecalis ATCC 29212 | 008 | 60 (100) | 0 (0) | 0 (0) |
| | | 009 | 60 (100) | 0 (0) | 0 (0) |
| | | 010 | 60 (100) | 0 (0) | 0 (0) |
| | | All | 180 (100) | 0 (0) | 0 (0) |
| | Staphylococcus aureus ATCC 29213 | 008 | 60 (100) | 0 (0) | 0 (0) |
| | | 009 | 60 (100) | 0 (0) | 0 (0) |
| | | 010 | 60 (100) | 0 (0) | 0 (0) |
| | | All | 180 (100) | 0 (0) | 0 (0) |
| | Staphylococcus epidermidis ATCC 12228 | 008 | 60 (100) | 0 (0) | 0 (0) |
| | | 009 | 60 (100) | 0 (0) | 0 (0) |
| | | 010 | 60 (100) | 0 (0) | 0 (0) |
| | | All | 180 (100) | 0 (0) | 0 (0) |
| | Staphylococcus saprophyticus ATCC 15305 | 008 | 60 (100) | 0 (0) | 0 (0) |
| | | 009 | 60 (100) | 0 (0) | 0 (0) |
| | | 010 | 60 (100) | 0 (0) | 0 (0) |
| | | All | 180 (100) | 0 (0) | 0 (0) |
| | Streptococcus agalactiae ATCC 12386 | 008 | 60 (100) | 0 (0) | 0 (0) |
| | | 009 | 60 (100) | 0 (0) | 0 (0) |
| | | 010 | 59 (98.3) | 0 (0) | 1 (1.7) |
| | | All | 179 (99.4) | 0 (0) | 1 (0.6) |
| | All Gram-Positive species (n=900) | 008 | 300 (100) | 0 (0) | 0 (0) |
| | | 009 | 300 (100) | 0 (0) | 0 (0) |
| | | 010 | 300 (100) | 0 (0) | 0 (0) |
| | | All | 899 (99.9) | 0 (0) | 1 (0.1) |
| All Species (n=1800) | 008 | 540 (90.0) | 60 (10.0) | 0 (0) | |
| | | 009 | 540 (90.0) | 60 (10.0) | 0 (0) |
| | | 010 | 539 (89.8) | 60 (10.0) | 1 (0.2) |
| | | All | 1619 (89.9) | 180 (10.0) | 1 (0.1) |

ATCC: American Type Culture Collection
Identified by VITEK MS as "Enterobacter cloacae/Enterobacter asburiae" in accordance with the labeling for the analyzer.

## b. Reproducibility of Copan Colibri with the Bruker MALDI Biotyper CA

Overall, for the Copan Colibri System in conjunction with the Bruker MALDI Biotyper CA, there was 88.1% agreement (1585/1800) between the reported High Confidence identification results (Log(Score) ≥ 2.00) and the expected identity of each colony in the Reproducibility Study (Gram-positive and Gram-negative species combined) (Table 4). However, for Gram-positive species, 179/900 colonies (19.9%) were only identified with Low Confidence (Log(Score) = 1.70-1.99), compared with 1/900 colonies (0.1%) of Gram-negative species. In addition, 31/900 Gram-positive colonies (3.4%) produced no identification result compared with 4/900 (0.4%) Gram-negative colonies. Nevertheless, no significant differences in agreement were observed using Fisher's Exact test between instruments and operators, and none of the samples in the study were incorrectly identified. Because the instructions for use require additional testing to confirm organism identity when a low confidence result or no identification is obtained, the study results were determined to be acceptable.

The reproducibility of the Copan Colibri System with Gram-positive and Gram-negative species is reported separately in the device labeling with instructions for the operator to

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perform additional testing to determine the identity of any isolate that yields either a Low Confidence identification result or no identification in accordance with the Bruker predicate device's labeling.

Table 4. Reproducibility of the Copan Colibri System with the Bruker MALDI Biotyper CA.

| Gram Stain Phenotype | Species/Strain | Colibri System | Bruker MALDI Biotyper CA Identification Result | | |
| --- | --- | --- | --- | --- | --- |
| | | | High Confidence | Low Confidence | None |
| Gram-Negative | Enterobacter cloacae1ATCC 13047 | 008 | 60 (100) | 0 (0) | 0 (0) |
| | | 009 | 60 (100) | 0 (0) | 0 (0) |
| | | 010 | 60 (100) | 0 (0) | 0 (0) |
| | | All | 180 (100) | 0 (0) | 0 (0) |
| | Escherichia coliATCC 25922 | 008 | 60 (100) | 0 (0) | 0 (0) |
| | | 009 | 58 (96.7) | 1 (1.7) | 1 (1.7) |
| | | 010 | 60 (100) | 0 (0) | 0 (0) |
| | | All | 178 (98.9) | 1 (0.6) | 1 (0.6) |
| | Klebsiella pneumoniaeATCC BAA 1705 | 008 | 60 (100) | 0 (0) | 0 (0) |
| | | 009 | 60 (100) | 0 (0) | 0 (0) |
| | | 010 | 60 (100) | 0 (0) | 0 (0) |
| | | All | 180 (100) | 0 (0) | 0 (0) |
| | Proteus mirabilisATCC 7002 | 008 | 60 (100) | 0 (0) | 0 (0) |
| | | 009 | 60 (100) | 0 (0) | 0 (0) |
| | | 010 | 60 (100) | 0 (0) | 0 (0) |
| | | All | 180 (100) | 0 (0) | 0 (0) |
| | Pseudomonas aeruginosaATCC 27853 | 008 | 59 (98.3) | 0 (0) | 1 (1.7) |
| | | 009 | 60 (100) | 0 (0) | 0 (0) |
| | | 010 | 58 (96.7) | 0 (0) | 2 (3.3) |
| | | All | 177 (98.3) | 0 (0) | 3 (1.7) |
| | All Gram-Negative species (n=900) | 008 | 299 (99.7) | 0 (0) | 1 (0.3) |
| | | 009 | 298 (99.3) | 1 (0.3) | 1 (0.3) |
| | | 010 | 298 (99.3) | 0 (0) | 2 (0.7) |
| | | All | 895 (99.4) | 1 (0.1) | 4 (0.4) |
| Gram-Positive | Enterococcus faecalisATCC 29212 | 008 | 47 (78.3) | 12 (20.0) | 1 (1.7) |
| | | 009 | 42 (70.00 | 18 (30.0) | 0 (0) |
| | | 010 | 50 (83.3) | 10 (16.7) | 0 (0) |
| | | All | 139 (77.2) | 40 (22.2) | 1 (0.6) |
| | Staphylococcus aureusATCC 29213 | 008 | 56 (93.3) | 4 (6.7) | 0 (0) |
| | | 009 | 56 (93.3) | 4 (6.7) | 0 (0) |
| | | 010 | 47 (78.3) | 13 (21.7) | 0 (0) |
| | | All | 159 (88.3) | 21 (11.7) | 0 (0) |
| | Staphylococcus epidermidisATCC 12228 | 008 | 43 (71.7) | 15 (25.0) | 2 (3.3) |
| | | 009 | 45 (75.0) | 12 (20.0) | 3 (5.0) |
| | | 010 | 41 (68.3) | 15 (25.0) | 4 (6.7) |
| | | All | 129 (71.7) | 42 (23.3) | 9 (5.0) |
| | Staphylococcus saprophyticusATCC 15305 | 008 | 48 (80.0) | 8 (13.3) | 4 (6.7) |
| | | 009 | 48 (80.0) | 8 (13.3) | 4 (6.7) |
| | | 010 | 47 (78.3) | 10 (16.7) | 3 (5.0) |
| | | All | 143 (79.4) | 26 (14.4) | 11 (6.1) |
| | Streptococcus agalactiaeATCC 12386 | 008 | 42 (70.0) | 15 (25.0) | 3 (5.0) |
| | | 009 | 40 (66.7) | 16 (26.7) | 4 (6.7) |
| | | 010 | 38 (63.3) | 19 (31.7) | 3 (5.0) |
| | | All | 120 (66.7) | 50 (27.8) | 10 (5.6) |
| | All Gram-Positive species (n=900) | 008 | 236 (78.7) | 54 (18.0) | 10 (3.3) |
| | | 009 | 232 (77.3) | 57 (19.0) | 11 (3.7) |
| | | 010 | 222 (74.0) | 68 (22.7) | 10 (3.3) |
| | | All | 690 (76.7) | 179 (19.9) | 31 (3.4) |
| All Species (n=1800) | 008 | 535 (89.2) | 54 (9.0) | 11 (1.8) | |
| | | 009 | 529 (88.2) | 59 (9.8) | 12 (2.0) |
| | | 010 | 521 (86.8) | 67 (11.2) | 12 (2.0) |
| | | All | 1585 (88.1) | 180 (10.0) | 35 (1.9) |

Note: The Reproducibility Study with the Bruker MALDI Biotyper CA was conducted using 48-spot reusable target plates

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ATCC: American Type Culture Collection.

1 Identified by the Bruker MALDI Biotyper CA as "Enterobacter cloacae complex" in accordance with the labeling for the analyzer.

# 2. Linearity:

Not applicable.

# 3. Analytical Specificity/Interference:

Refer to Section VII (A)(4).

# 4. Accuracy (Instrument):

# Colony Picking Accuracy

The ability of the Copan Colibri Preparation Station to pick appropriate colonies designated by the operator based on the digital images obtained by the Copan Colibri Vision System was evaluated in separate studies for each of the two downstream MALDI-TOF MS analyzers, as described below.

In each study, three different Colibri Systems, with three different operators, were used to prepare targets from isolated colonies of mixed cultures on both whole and bi-plates of representative "on panel" Gram-positive and Gram-negative bacterial species (Table 5). Once automated processing of the colonies was completed, the targets were removed from the Colibri Preparation Station and the appropriate calibration spots were applied manually before transfer to the MALDI-TOF MS analyzers.

Table 5. Organism mixtures and culture media use for evaluation of colony picking accuracy

| Mixture | Species | Culture Medium | |
| --- | --- | --- | --- |
| | | VITEK MS | MALDI Biotyper CA |
| 1 | Escherichia coli ATCC 25922
Klebsiella pneumoniae ATCC BAA 1705
Staphylococcus aureus ATCC 29213 | Trypticase Soy Agar + 5% Sheep Blood | Trypticase Soy Agar + 5% Sheep Blood |
| 2 | Staphylococcus aureus ATCC 29213
Enterococcus faecalis ATCC 29212
Streptococcus agalactiae ATCC 12386 | Trypticase Soy Agar + 5% Sheep Blood | Columbia Agar + 5% Sheep Blood with colistin and nalidixic acid (CNA) |
| 3 | Escherichia coli ATCC 25922
Klebsiella pneumoniae ATCC BAA 1705
Proteus mirabilis ATCC 7002 | MacConkey Agar | MacConkey Agar |
| 4 | Escherichia coli ATCC 25922
Klebsiella pneumoniae ATCC BAA 1705
Staphylococcus aureus ATCC 29213 | Trypticase Soy Agar + 5% Sheep Blood/MacConkey Agar Bi-plate | Trypticase Soy Agar + 5% Sheep Blood/MacConkey Agar Bi-plate |
| 5 | Escherichia coli ATCC 25922
Klebsiella pneumoniae ATCC BAA 1705
Staphylococcus aureus ATCC 29213
Staphylococcus epidermidis ATCC 12228 | N/A | Columbia Agar + 5% Sheep Blood/MacConkey Agar Bi-plate |

Processed culture plates were inspected manually in comparison to the corresponding digital

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images to determine whether the designated colonies had been picked. Additional colonies were also picked manually and used to prepare targets for mass spectrometry analysis. The results obtained with targets prepared by the Colibri System were compared to the expected identity of each organism, i.e., the known species identifications of the reference isolates obtained from ATCC, as well as to the identification results obtained with manually prepared targets, as described below. Colony picking accuracy of Copan Colibri with the bioMérieux VITEK MS and with the Bruker MALDI Biotyper CA are outlined below in sections $a$ and $b$ , respectively.

# a. Colony Picking Accuracy for bioMérieux VITEK MS

Three Colibri System instruments were used in this study. A total of 1390 colonies were picked by the three instruments to prepare target slides for the VITEK MS. Of those, 924 and 466 colonies were from whole plates and bi-plates, respectively. All colonies (100%) were successfully picked by visual inspection. For all species excluding $E$ faecalis, $\geq 95\%$ of designated colonies were correctly identified with good confidence (Confidence Value $\geq 60\%$ ) compared to the expected identity.

Results were stratified according to species, Colibri System and whole plate (Table 6) or bi-plate configuration (Table 7). The Copan Colibri System performed equivalently between the three instruments and between whole plates and bi-plates. For the combined data for whole plates and bi-plates, 1368/1390 (98.4%) produced Good Confidence identification results that agreed with the expected organism identity (Table 8). The percent agreement for Gram-negative species was higher than for Gram-positive organisms (100% vs 96.5%), although no incorrect identification results were reported for any organism. In comparison, manual preparation of VITEK MS target slides yielded 1351/1390 (97.2%) Good Confidence identification results that agreed with the expected organism identity. When used in conjunction with the VITEK MS, the accuracy of colony picking by the Copan Colibri System was therefore determined to be acceptable.

Table 6. Comparison of expected identifications to bioMérieux VITEK MS results from target slides prepared on the Copan Colibri System from whole plates.

| Expected Identification | Colibri System | N1 | VITEK MS Identification Result (%)2 | | |
| --- | --- | --- | --- | --- | --- |
| | | | Good Confidence | Low Discrimination | No Identification |
| Escherichia coli | 008 | 46 | 46 (100) | 0 (0) | 0 (0) |
| | 009 | 50 | 50 (100) | 0 (0) | 0 (0) |
| | 010 | 50 | 50 (100) | 0 (0) | 0 (0) |
| | Overall | 146 | 146 (100) | 0 (0) | 0 (0) |
| Klebsiella pneumoniae | 008 | 44 | 44 (100) | 0 (0) | 0 (0) |
| | 009 | 52 | 52 (100) | 0 (0) | 0 (0) |
| | 010 | 50 | 50 (100) | 0 (0) | 0 (0) |
| | Overall | 146 | 146 (100) | 0 (0) | 0 (0) |
| Proteus mirabilis | 008 | 50 | 50 (100) | 0 (0) | 0 (0) |
| | 009 | 56 | 56 (100) | 0 (0) | 0 (0) |
| | 010 | 52 | 52 (100) | 0 (0) | 0 (0) |
| | Overall | 158 | 158 (100) | 0 (0) | 0 (0) |
| Enterococcus faecalis | 008 | 50 | 41 (82.0) | 1 (2.0) | 8 (16.0) |
| | 009 | 54 | 48 (88.9) | 0 (0) | 6 (11.1) |
| | 010 | 64 | 63 (98.4) | 0 (0) | 1 (1.6) |

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| Expected Identification | Colibri System | N1 | VITEK MS Identification Result (%)2 | | |
| --- | --- | --- | --- | --- | --- |
| | | | Good Confidence | Low Discrimination | No Identification |
| | Overall | 168 | 152 (90.5) | 1 (0.6) | 15 (8.9) |
| Staphylococcus aureus | 008 | 48 | 48 (100) | 0 (0) | 0 (0) |
| | 009 | 48 | 48 (100) | 0 (0) | 0 (0) |
| | 010 | 48 | 48 (100) | 0(0) | 0 (0) |
| | Overall | 144 | 144 (100) | 0 (0) | 0 (0) |
| Streptococcus agalactiae | 008 | 50 | 48 (96.0) | 0 (0) | 2 (4.0) |
| | 009 | 56 | 54 (96.4) | 0 (0) | 2 (3.6) |
| | 010 | 56 | 54 (96.4) | 0 (0) | 2 (3.6) |
| | Overall | 162 | 156 (96.3) | 0 (0) | 6 (3.7) |
| All Species | 008 | 288 | 277 (96.2) | 1 (0.3) | 10 (3.5) |
| | 009 | 316 | 308 (97.5) | 0 (0) | 8 (2.5) |
| | 010 | 320 | 317 (99.1) | 0 (0) | 3 (0.9) |
| | Overall | 924 | 902 (97.6) | 1 (0.1) | 21 (2.3) |

1 Number of colonies processed.
2 No incorrect identifications were reported.

Table 7. Comparison of expected identifications to bioMérieux VITEK MS results from target slides prepared on the Copan Colibri System from bi-plates.

| Expected Identification | Colibri System | N1 | VITEK MS Identification Result (%)2 | | |
| --- | --- | --- | --- | --- | --- |
| | | | Good Confidence | Low Discrimination | No Identification |
| Escherichia coli | 008 | 46 | 46 (100) | 0 (0) | 0 (0) |
| | 011 | 58 | 58 (100) | 0 (0) | 0 (0) |
| | 013 | 50 | 50 (100) | 0 (0) | 0 (0) |
| | Overall | 154 | 154 (100) | 0 (0) | 0 (0) |
| Klebsiella pneumoniae | 008 | 44 | 44 (100) | 0 (0) | 0 (0) |
| | 011 | 72 | 72 (100) | 0 (0) | 0 (0) |
| | 013 | 48 | 48 (100) | 0 (0) | 0 (0) |
| | Overall | 164 | 164 (100) | 0 (0) | 0 (0) |
| Staphylococcus aureus | 008 | 56 | 56 (100) | 0 (0) | 0 (0) |
| | 011 | 48 | 48 (100) | 0 (0) | 0 (0) |
| | 013 | 44 | 44 (100) | 0 (0) | 0 (0) |
| | Overall | 148 | 148 (100) | 0 (0) | 0 (0) |
| All Species | 008 | 146 | 146 (100) | 0 (0) | 0 (0) |
| | 011 | 178 | 178 (100) | 0 (0) | 0 (0) |
| | 013 | 142 | 142 (100) | 0 (0) | 0 (0) |
| | Overall | 466 | 466 (100) | 0 (0) | 0 (0) |

1 Number of colonies processed.
2 No incorrect identifications were reported.

Table 8. Summary of bioMérieux VITEK MS results from target slides prepared on the Copan Colibri System compared to manual preparation.

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| Organism | N1 | VITEK MS Identification Result (%)2 | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- |
| | | Good Confidence | | Low Discrimination | | No Identification | |
| | | Colibrí | Manual | Colibrí | Manual | Colibrí | Manual |
| Escherichia coli | 300 | 300 (100) | 298 (99.3) | 0 (0) | 0 (0) | 0 (0) | 2 (0.7) |
| Klebsiella pneumoniae | 310 | 310 (100) | 300 (96.8) | 0 (0) | 0 (0) | 0 (0) | 10 (3.2) |
| Proteus mirabilis | 158 | 158 (100) | 154 (97.5) | 0 (0) | 0 (0) | 0 (0) | 4 (2.5) |
| All Gram-Negative Species | 768 | 768 (100) | 752 (97.9) | 0 (0) | 0 (0) | 0 (0) | 16 (2.1) |
| Enterococcus faecalis | 168 | 152 (90.5) | 156 (92.9) | 1 (2.0) | 0 (0) | 15 (8.9) | 12 (7.1) |
| Staphylococcus aureus | 292 | 292 (100) | 291 (99.7) | 0 (0) | 0 (0) | 0 (0) | 1 (0.3) |
| Streptococcus agalactiae | 162 | 156 (96.3) | 152 (93.8) | 0 (0) | 1 (0.6) | 6 (3.7) | 9 (5.6) |
| All Gram-Positive Species | 622 | 600 (96.5) | 599 (96.3) | 1 (0.2) | 1 (0.2) | 21 (3.4) | 22 (3.5) |
| Overall | 1390 | 1368 (98.4) | 1351 (97.2) | 1 (0.1) | 1 (0.1) | 21 (1.5) | 38 (2.7) |

1 Number of colonies processed.
2 No incorrect identifications were reported.

## b. Colony Picking Accuracy for Bruker MALDI Biotyper CA

Three Colibrí System instruments were used in this study. A total of 1690 colonies were picked by the three instruments to prepare 48-spot reusable targets for the MALDI Biotyper CA. Of these, 898 and 792 colonies were from whole plates and bi-plates, respectively. All colonies (100%) were successfully picked by visual inspection.

Identification results were stratified according to species, Colibrí System and whole plate (Table 9) or bi-plate configuration (Table 10). The Copan Colibrí System performed equivalently between instruments and between whole plates and bi-plates. For the combined data for whole plates and bi-plates, 1534/1690 (90.8%) produced high confidence identification results that agreed with the expected organism identity (Table 11). The percent agreement for Gram-negative species was higher than for Gram-positive organisms [873/878 (99.4%) vs. 661/812 (81.4%)], although no incorrect identification results were reported for any organism. Manual preparation of MALDI targets yielded 1604/1690 (94.9%) high confidence identification results that agreed with the expected organism identity, with 99.2% agreement for Gram-negatives and 90.3% agreement for Gram-positives.

Table 9. Comparison of expected identifications to Bruker MALDI Biotyper CA results from targets prepared on the Copan Colibri System from whole plates.

| Expected Identification | Colibri System | N1 | MALDI Biotyper CA Identification Result (%)2 | | |
| --- | --- | --- | --- | --- | --- |
| | | | High Confidence | Low Confidence | No Identification |
| Escherichia coli | 008 | 40 | 40 (100) | 0 (0) | 0 (0) |
| | 009 | 40 | 39 (97.5) | 1 (2.5) | 0 (0) |
| | 010 | 64 | 61 (95.3) | 3 (4.7) | 0 (0) |
| | Overall | 144 | 140 (97.2) | 4 (2.8) | 0 (0) |
| Klebsiella pneumoniae | 008 | 46 | 46 (100) | 0 (0) | 0 (0) |

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Table 10. Comparison of expected identifications to Bruker MALDI Biotyper CA results from targets prepared on the Copan Colibri System from bi-plates.

| Expected Identification | Colibri System | N1 | MALDI Biotyper CA Identification Result (%)2 | | |
| --- | --- | --- | --- | --- | --- |
| | | | High Confidence | Low Confidence | No Identification |
| Escherichia coli | 008 | 70 | 70 (100) | 0 (0) | 0 (0) |
| | 011 | 84 | 84 (100) | 0 (0) | 0 (0) |
| | 013 | 82 | 81 (98.8) | 1 (1.2) | 0 (0) |
| | Overall | 236 | 235 (99.6) | 1 (0.4) | 0 (0) |
| Klebsiella pneumoniae | 008 | 70 | 70 (100) | 0 (0) | 0 (0) |
| | 011 | 70 | 70 (100) | 0 (0) | 0 (0) |
| | 013 | 48 | 48 (100) | 0 (0) | 0 (0) |
| | Overall | 188 | 188 (100) | 0 (0) | 0 (0) |
| Staphylococcus aureus | 008 | 74 | 61 (82.4) | 13 (17.6) | 0 (0) |
| | 011 | 70 | 62 (88.6) | 8 (11.4) | 0 (0) |
| | 013 | 76 | 71 (93.4) | 5 (6.6) | 0 (0) |
| | Overall | 220 | 194 (88.2) | 26 (11.8) | 0 (0) |
| Staphylococcus epidermidis | 008 | 50 | 39 (78.0) | 11 (22.0) | 0 (0) |
| | 011 | 50 | 41 (82.0) | 9 (18.0) | 0 (0) |

1 Number of colonies processed.
2 No incorrect identifications were reported.

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| Expected Identification | Colibri System | N1 | MALDI Biotyper CA Identification Result (%)2 | | |
| --- | --- | --- | --- | --- | --- |
| | | | High Confidence | Low Confidence | No Identification |
| | 013 | 48 | 38 (79.2) | 10 (20.8) | 0 (0) |
| | Overall | 148 | 118 (79.7) | 30 (20.3) | 0 (0) |
| All Species | 008 | 264 | 240 (90.9) | 24 (9.1) | 0 (0) |
| | 011 | 274 | 257 (93.8) | 17 (6.2) | 0 (0) |
| | 013 | 254 | 238 (93.7) | 16 (6.3) | 0 (0) |
| | Overall | 792 | 735 (92.8) | 57 (7.2) | 0 (0) |

1 Number of colonies processed.
2 No incorrect identifications were reported.

Table 11. Summary of Bruker MALDI Biotyper CA results from targets prepared on the Copan Colibri System compared to manual preparation.

| Organism | N1 | MALDI Biotyper CA Identification Result (%)2 | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- |
| | | High Confidence | | Low Confidence | | No Identification | |
| | | Colibri | Manual | Colibri | Manual | Colibri | Manual |
| Escherichia coli | 380 | 375(98.7) | 377(99.2) | 5(1.3) | 3(0.8) | 0(0) | 0(0) |
| Klebsiella pneumoniae | 330 | 330(100) | 326(98.8) | 0(0) | 1(0.3) | 0(0) | 3(0.9) |
| Proteus mirabilis | 168 | 168(100) | 168(100) | 0(0) | 0(0) | 0(0) | 0(0) |
| All Gram-Negative Species | 878 | 873(99.4) | 871(99.2) | 5(0.6) | 4(0.5) | 0(0) | 3(0.3) |
| Enterococcus faecalis | 150 | 122(81.3) | 144(96.0) | 22(14.7) | 4(2.7) | 6(4.0) | 2(1.3) |
| Staphylococcus aureus | 364 | 316(86.8) | 334(91.8) | 48(13.2) | 30(8.2) | 0(0) | 0(0) |
| Staphylococcus epidermidis | 148 | 118(79.7) | 117(79.1) | 30(20.3) | 31(20.9) | 0(0) | 0(0) |
| Streptococcus agalactiae | 150 | 105(70.0) | 138(92.0) | 26(17.3) | 8(5.3) | 19(12.7) | 4(2.7) |
| All Gram-Positive Species | 812 | 661(81.4) | 733(90.3) | 126(15.5) | 73(9.0) | 25(3.1) | 6(0.7) |
| Overall | 1690 | 1534(90.8) | 1604(94.9) | 131(7.7) | 77(4.5) | 25(1.5) | 9(0.4) |

1 Number of colonies processed.
2 No incorrect identifications were reported.

The accuracy of colony picking by the Copan Colibri System for preparation of targets for the Bruker MALDI Biotyper CA was determined to be acceptable because, the designated colonies were picked accurately, and no incorrect identification results were reported. In addition, according to the Instructions for Use of the Bruker MALDI Biotyper CA System, if a low-confidence identification or a no identification result is obtained using the Direct Transfer (DT) Sample Preparation Procedure, as implemented with the Copan Colibri System, the operator is instructed to repeat testing of the isolate manually using the extended Direct Transfer (eDT) or Extraction (Ext) Sample Preparation Procedure.

# Accuracy of Bacterial Identification (Inclusivity)

The accuracy of MALDI-TOF MS organism identification with targets prepared by the

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Copan Colibri System was evaluated by testing representative strains of clinically relevant Gram-positive and Gram-negative bacteria that exhibited a range of colony characteristics in terms of size, consistency and morphology. Isolates were obtained from the American Type Culture Collection (ATCC) as well as other sources (i.e., clinical isolates). The identity of the clinical isolates was confirmed using the bioMérieux VITEK MS or VITEK 2. Results obtained using Copan Colibri and the VITEK MS or Bruker MALDI Biotyper CA analyzers were compared to the expected organism identity as well as results from manually prepared samples. Accuracy of bacterial identification by Copan Colibri with the bioMérieux VITEK MS and with the Bruker MALDI Biotyper CA are outlined below in sections $a$ and $b$ , respectively.

# a. Accuracy of Bacterial Identification with the bioMérieux VITEK MS

A total of 123 strains from 29 "on panel" bacterial species, i.e. represented in the FDA cleared MS reference database, were cultured on appropriate media and under appropriate conditions. Targets were prepared in duplicate, using 1 colony per spot and 2 spots per isolate. The results of the study with the VITEK MS are summarized in Tables 12 and 13, for Gram-negative and Gram-positive species, respectively, and stratified according to species. The percentage of target spots correctly identified with Good Confidence discrimination (Confidence Value $\geq 60\%$ ) in comparison to the expected identity was calculated.

For Gram-negative species, 258/308 colonies $(83.8\%)$ had Good Confidence identification results that matched the expected organism identity. Twenty-three colonies of Enterobacter cloacae complex were reported with Low Discrimination as $E.$ cloacae/E. asburiae in accordance with the labeling for the VITEK MS analyzer. In addition, 24 colonies of $P.$ vulgaris processed by the Colibri System were identified with Low Discrimination as $P.$ vulgaris/P. penneri in accordance with the labeling for the VITEK MS analyzer, which also agreed with the organism identity obtained by manual processing of 23/24 colonies from the same culture plates.

For Gram-positive species, 78/84 colonies $(92.9\%)$ were identified correctly with Good Confidence values.

None of the colonies of either the Gram-positive or Gram-negative species that were processed by the Copan Colibri System produced an incorrect identification result.

The results obtained from manual and automated preparation of targets on the Copan Colibri System are compared in Table 14.

When low discrimination occurred, results were as expected in accordance with labeling and there were no misidentifications. Therefore, the accuracy of bacterial identification obtained using the Colibri System in conjunction with the VITEK MS analyzer was therefore determined to be acceptable.

Table 12. Summary of results from representative Gram-negative bacteria using the Copan Colibri System in conjunction with the bioMérieux VITEK MS.

| Expected Organism Identity | Culture Medium | Strains Tested | Colonies Picked1 | VITEK MS Identification Result | | | % Agreement2 | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | | | Good Confidence | Low Discrimination | No ID | Culture Medium | Species/Group |
| Acinetobacter baumannii | MAC | 6 | 12 | 12 | -- | -- | 100 | 100 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Bacteroides fragilis | TSA | 1 | 2 | 2 | -- | -- | 100 | 100 |

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| Expected Organism Identity | Culture Medium | Strains Tested | Colonies Picked^{1} | VITEK MS Identification Result | | | % Agreement^{2} | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | | | Good Confidence | Low Discrimination | No ID | Culture Medium | Species/Group |
| Citrobacter koseri | MAC | 6 | 12 | 12 | -- | -- | 100 | 100 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Eikenella corrodens | TSA | 1 | 2 | 2 | -- | -- | 100 | 100 |
| Enterobacter aerogenes | MAC | 6 | 12 | 11 | -- | 1 | 91.7 | 95.8 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Enterobacter cloacae complex | MAC | 6 | 12 | -- | 11^{3} | 1 | 91.7^{3} | 95.8^{3} |
| | TSA | 6 | 12 | -- | 12^{3} | -- | 100^{3} | |
| Escherichia coli | MAC | 6 | 12 | 12 | -- | -- | 100 | 100 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Haemophilus influenzae | CHO | 2 | 4 | 4 | -- | -- | 100 | 100 |
| Klebsiella oxytoca | MAC | 6 | 12 | 12 | -- | -- | 100 | 100 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Klebsiella pneumoniae | MAC | 6 | 12 | 12 | -- | -- | 100 | 95.8 |
| | TSA | 6 | 12 | 11 | -- | 1 | 91.7 | |
| Moraxella catarrhalis | CHO | 1 | 2 | 2 | -- | -- | 100 | 100 |
| | TSA | 1 | 2 | 2 | -- | -- | 100 | |
| Morganella morganii | MAC | 4 | 8 | 8 | -- | -- | 100 | 100 |
| | TSA | 4 | 8 | 8 | -- | -- | 100 | |
| Neisseria gonorrhoeae | CHO | 2 | 4 | 4 | -- | -- | 100 | 100 |
| Neisseria meningitidis | CHO | 1 | 2 | 2 | -- | -- | 100 | 100 |
| Proteus mirabilis | MAC | 6 | 12 | 12 | -- | -- | 100 | 100 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Proteus vulgaris | MAC | 6 | 12 | -- | 12^{4} | -- | 100^{4} | 100^{4} |
| | TSA | 6 | 12 | -- | 12^{4} | -- | 100^{4} | |
| Pseudomonas aeruginosa | MAC | 6 | 12 | 12 | -- | -- | 100 | 100 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Salmonella typhimurium | MAC | 2 | 4 | 4^{5} | -- | -- | 100 | 100 |
| | TSA | 2 | 4 | 4^{5} | -- | -- | 100 | |
| Serratia marcescens | MAC | 4 | 8 | 8 | -- | -- | 100 | 100 |
| | TSA | 4 | 8 | 8 | -- | -- | 100 | |
| Stenotrophomonas maltophilia | MAC | 2 | 4 | 4 | -- | -- | 100 | 100 |
| | TSA | 2 | 4 | 4 | -- | -- | 100 | |
| Vibrio parahaemolyticus | TSA | 1 | 2 | 2 | -- | -- | 100 | 100 |
| | Total (%) | 154 | 308 (100) | 258 (83.8) | 47 (15.3) | 3 (1.0) | | 305 (99.0)^{6} |

No ID: No identification; CHO: Chocolate Agar; MAC: MacConkey Agar; TSA: Trypticase Soy Agar + 5% Sheep Blood
1 2 colonies of each strain were designated for picking by the Colibri System and each was used to prepare a separate target spot.
2 Agreement with Expected Organism Identity with Good Confidence.
3 Reported as Enterobacter cloacae/Enterobacter asburiae in accordance with VITEK MS device labeling so low discrimination results for this strain are included in the calculation of agreement.
4 Reported as Proteus vulgaris/Proteus penneri in accordance with VITEK MS device labeling so low discrimination results for this strain are included in the calculation of agreement (23/24 colonies processed manually from the same culture plates were also reported as P. vulgaris/P. penneri).
5 Reported as Salmonella group.
6 Calculation of agreement includes the 47/48 slashline results for Enterobacter cloacae and Proteus vulgaris that are reported

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with low discrimination in accordance with the labeling for the VITEK MS analyzer.

Table 13. Summary of results from representative Gram-positive bacteria using the Copan Colibri System in conjunction with the bioMérieux VITEK MS.

| Expected Organism Identity | Strains Tested | Colonies Picked2 | VITEK MS Identification Result | | | % Agreement3 |
| --- | --- | --- | --- | --- | --- | --- |
| | | | Good Confidence | Low Discrimination | No ID | |
| Enterococcus faecium | 6 | 12 | 11 | -- | 1 | 91.7 |
| Enterococcus faecalis | 6 | 12 | 11 | -- | 1 | 91.7 |
| Listeria monocytogenes | 2 | 4 | 4 | -- | -- | 100 |
| Staphylococcus aureus | 6 | 12 | 12 | -- | -- | 100 |
| Staphylococcus epidermidis | 6 | 12 | 12 | -- | -- | 100 |
| Staphylococcus saprophyticus | 4 | 8 | 6 | -- | 2 | 75.0 |
| Streptococcus agalactiae | 6 | 164 | 13 | -- | 3 | 81.3 |
| Streptococcus pyogenes | 4 | 8 | 7 | -- | 1 | 87.5 |
| Total (%) | 40 | 84 (100) | 76 (90.5) | 0 (0.0) | 8 (9.5) | |

No ID: No Identification
1 All Gram-positive organisms evaluated with VITEK MS were cultured on Trypticase Soy Agar + 5% Sheep Blood.
2 2 colonies of each strain were designated for picking by the Colibri System and each was used to prepare a separate target spot.
3 Agreement with Expected Organism Identity with Good Confidence (60 - 99.9%).
4 1 strain was tested twice (total 4 colonies, 3 of which produced the expected identification result with Good Confidence).

Table 14. Comparison of bioMérieux VITEK MS results from targets prepared on the Copan Colibri System or manually.

| Organism | Colonies Picked | VITEK MS Identification Result | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- |
| | | Good Confidence | | Low Discrimination | | No Identification | |
| | | Colibrí | Manual | Colibrí | Manual | Colibrí | Manual |
| Gram-Negative | 308 | 2581(83.8) | 238(77.3) | 47(15.3) | 43(14.0) | 3(1.0) | 27(8.8) |
| Gram-Positive | 84 | 76(90.5) | 78(92.9) | 0(0.0) | 0(0.0) | 8(9.5) | 6(7.1) |
| Total | 392 | 3341(85.2) | 316(80.6) | 47(12.0) | 43(11.0) | 11(2.8) | 33(8.4) |

1 Percent agreement for automated preparation is $99.0\%$ for Gram-negative organisms and $97.2\%$ in total when the 47 low discrimination results for Enterobacter cloacae and Proteus vulgaris are included. This is in accordance with the VITEK MS device labeling, which cannot differentiate between Enterobacter cloacae and Enterobacter asburiae or Proteus vulgaris and Proteus penneri and reports slashline results with low discrimination.

# b. Accuracy of Bacterial Identification with the Bruker MALDI Biotyper CA

A total of 124 strains from 30 "on panel" bacterial species, i.e. represented in the validated MS reference database, were cultured on appropriate media and under appropriate conditions. Targets were prepared in duplicate, using 1 colony per spot and 2 spots per isolate. Testing was performed separately with both the reusable (48-spot) and disposable (96-spot) targets. Results obtained from the Inclusivity Study with the Bruker MALDI Biotyper CA 48-spot targets are summarized in Tables 15 and 16 and those obtained with 96-spot targets are shown in Tables 17 and 18, stratified according to species. The percentage of target spots correctly identified with High Confidence $(\mathrm{Log}(\mathrm{Score})\mathrm{Value}\geq 2.00)$ in comparison to the expected identity was calculated.

For Gram-negative species, the expected organism identity was reported with High

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Confidence Log(Score) values for 308/312 colonies (98.7%) spotted onto 48-spot reusable targets and 296/312 colonies (94.9%) that were processed using 96-spot disposable targets. In contrast, a lower proportion of colonies of Gram-positive organisms yielded High Confidence identification results that agreed with the expected identity. With 48-spot reusable targets, 128/156 colonies (82.1%) were identified with High Confidence Log(Score) values, as were 105/156 colonies (67.3%) spotted on 96-spot disposable targets.

A comparison of results from targets prepared on 48-spot slides on the Copan Colibri System and from those prepared manually is shown in Table 19.

The performance of the Copan Colibri System for preparation of Gram-positive target organisms for the Bruker MALDI Biotyper CA is lower when compared to manual preparation. However, no incorrect identification results were reported, and instructions will be included in the Copan Colibri System Operator Manual for the operator to repeat testing of the isolate manually using the extended Direct Transfer (eDT) or Extraction (Ext) Sample Preparation Procedure if a low-confidence identification or no identification result is obtained. This is consistent with the Instructions for Use of the Bruker MALDI Biotyper CA System. Therefore, the results of the Inclusivity Study with the Bruker MALDI Biotyper CA were determined to be acceptable.

Table 15. Summary of results from representative Gram-negative bacteria using the Copan Colibri System in conjunction with the Bruker MALDI Biotyper CA using 48-spot targets.

| Expected Organism Identity | Culture Medium | Strains Tested | Colonies Picked3 | MALDI Biotyper CA Identification Result | | | % Agreement1 | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | | | Confidence | | No ID2 | Culture Medium | Species/Group |
| | | | | High | Low | | | |
| Acinetobacter baumannii | MAC | 6 | 12 | 11 | 1 | -- | 91.7 | 95.8 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Bacteroides fragilis | TSA | 1 | 2 | 2 | -- | -- | 100 | 100 |
| Bordetella pertussis | BG | 1 | 2 | 2 | -- | -- | 100 | 100 |
| Citrobacter koseri | MAC | 6 | 12 | 12 | -- | -- | 100 | 100 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Eikenella corrodens | TSA | 2 | 4 | 4 | -- | -- | 100 | 100 |
| Enterobacter aerogenes | MAC | 6 | 12 | 12 | -- | -- | 100 | 100 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Enterobacter cloacae complex | MAC | 6 | 12 | 12 | -- | -- | 100 | 100 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Escherichia coli | MAC | 6 | 12 | 10 | 2 | -- | 83.3 | 91.7 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Haemophilus influenzae | CHO | 2 | 4 | 4 | -- | -- | 100 | 100 |
| Klebsiella oxytoca | MAC | 6 | 12 | 12 | -- | -- | 100 | 100 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Klebsiella pneumoniae | MAC | 6 | 12 | 12 | -- | -- | 100 | 100 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Moraxella catarrhalis | CHO | 1 | 2 | 2 | -- | -- | 100 | 100 |
| | TSA | 1 | 2 | 2 | -- | -- | 100 | |
| Morganella morganii | MAC | 4 | 8 | 8 | -- | -- | 100 | 100 |

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| Expected Organism Identity | Culture Medium | Strains Tested | Colonies Picked3 | MALDI Biotyper CA Identification Result | | | % Agreement1 | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | | | Confidence | | No ID2 | Culture Medium | Species/Group |
| | | | | High | Low | | | |
| | TSA | 4 | 8 | 8 | -- | -- | 100 | |
| Neisseria gonorrhoeae | CHO | 2 | 4 | 4 | -- | -- | 100 | 100 |
| Neisseria meningitidis | CHO | 1 | 2 | 2 | -- | -- | 100 | 100 |
| Proteus mirabilis | MAC | 6 | 12 | 12 | -- | -- | 100 | 100 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Proteus vulgaris | MAC | 6 | 12 | 11 | 1 | -- | 91.7 | 95.8 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Pseudomonas aeruginosa | MAC | 6 | 12 | 12 | -- | -- | 100 | 100 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Salmonella sp. | MAC | 2 | 4 | 4 | -- | -- | 100 | 100 |
| | TSA | 2 | 4 | 4 | -- | -- | 100 | |
| Serratia marcescens | MAC | 4 | 8 | 8 | -- | -- | 100 | 100 |
| | TSA | 4 | 8 | 8 | -- | -- | 100 | |
| Stenotrophomonas maltophilia | MAC | 2 | 4 | 4 | -- | -- | 100 | 100 |
| | TSA | 2 | 4 | 4 | -- | -- | 100 | |
| Vibrio parahaemolyticus | TSA | 1 | 2 | 2 | -- | -- | 100 | 100 |
| | Total (%) | 156 | 312 (100) | 308 (98.7) | 4 (1.3) | 0 (0.0) | | |

No ID: No identification; BG: Bordet Gengou Agar + 15% Sheep Blood; CHO: Chocolate Agar; MAC: MacConkey Agar; TSA: Trypticase Soy Agar + 5% Sheep Blood
1 Agreement with Expected Organism Identity with High Confidence Log(Score).
2 Includes reports of "No Identification" and "No peaks".
3 2 colonies of each strain were designated for picking by the Colibri System and each was used to prepare a separate target spot.

Table 16. Summary of results from representative Gram-positive bacteria using the Copan Colibri System in conjunction with the Bruker MALDI Biotyper CA using 48-spot targets.

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Table 17. Summary of results from representative Gram-negative bacteria using the Copan Colibri System in conjunction with the Bruker MALDI Biotyper CA using 96-spot targets.

| Expected Organism Identity | Culture Medium | Strains Tested | Colonies Picked3 | MALDI Biotyper CA Identification Result | | | % Agreement1 | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | | | Confidence | | No ID2 | Culture Medium | Species/Group |
| | | | | High | Low | | | |
| Enterococcus faecalis | TSA | 6 | 12 | 9 | 2 | 1 | 75.0 | 79.2 |
| | CNA | 6 | 12 | 10 | 2 | -- | 83.3 | |
| Enterococcus faecium | TSA | 6 | 12 | 10 | 2 | -- | 83.3 | 87.5 |
| | CNA | 6 | 12 | 11 | 1 | -- | 91.7 | |
| Listeria monocytogenes | TSA | 2 | 4 | 4 | -- | -- | 100 | 100 |
| Staphylococcus aureus | TSA | 6 | 12 | 9 | 3 | -- | 75.0 | 87.5 |
| | CNA | 6 | 12 | 12 | -- | -- | 100 | |
| Staphylococcus epidermidis | TSA | 6 | 12 | 9 | 3 | -- | 75.0 | 75.0 |
| | CNA | 6 | 12 | 9 | 3 | -- | 75.0 | |
| Staphylococcus saprophyticus | TSA | 4 | 8 | 8 | -- | -- | 100 | 75.0 |
| | CNA | 4 | 8 | 4 | 2 | 2 | 50.0 | |
| Streptococcus agalactiae | TSA | 6 | 12 | 9 | 2 | 1 | 75.0 | 79.2 |
| | CNA | 6 | 12 | 10 | -- | 2 | 83.3 | |
| Streptococcus pyogenes | TSA | 4 | 8 | 7 | 1 | -- | 87.5 | 87.5 |
| | CNA | 4 | 8 | 7 | 1 | -- | 87.5 | |
| | Total (%) | 78 | 156 (100) | 128 (82.1) | 22 (14.1) | 6 (3.8) | | |

No ID: No identification; CNA: Columbia Agar + 5% Sheep Blood with colistin and nalidixic acid (CNA); TSA: Trypticase Soy Agar +5% Sheep Blood
1 Agreement with Expected Organism Identity with High Confidence Log(Score).
2 Includes reports of "No Identification" and "No peaks".
3 2 colonies of each strain were designated for picking by the Colibri System and each was used to prepare a separate target spot.

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| Expected Organism Identity | Culture Medium | Strains Tested | Colonies Picked3 | MALDI Biotyper CA Identification Result | | | % Agreement1 | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | | | Confidence | | No ID2 | Culture Medium | Species/Group |
| | | | | High | Low | | | |
| Klebsiella oxytoca | MAC | 6 | 12 | 8 | 1 | 3 | 66.7 | 79.2 |
| | TSA | 6 | 12 | 11 | 1 | -- | 91.7 | |
| Klebsiella pneumoniae | MAC | 6 | 12 | 10 | 1 | 1 | 83.3 | 91.7 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Moraxella catarrhalis | CHO | 1 | 2 | 2 | -- | -- | 100 | 100 |
| | TSA | 1 | 2 | 2 | -- | -- | 100 | |
| Morganella morganii | MAC | 4 | 8 | 8 | -- | -- | 100 | 100 |
| | TSA | 4 | 8 | 8 | -- | -- | 100 | |
| Neisseria gonorrhoeae | CHO | 2 | 4 | 4 | -- | -- | 100 | 100 |
| Neisseria meningitidis | CHO | 1 | 2 | 2 | -- | -- | 100 | 100 |
| Proteus mirabilis | MAC | 6 | 12 | 11 | -- | 1 | 91.7 | 95.8 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Proteus vulgaris | MAC | 6 | 12 | 12 | -- | -- | 100 | 100 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Pseudomonas aeruginosa | MAC | 6 | 12 | 12 | -- | -- | 100 | 100 |
| | TSA | 6 | 12 | 12 | -- | -- | 100 | |
| Salmonella spp. | MAC | 2 | 4 | 4 | -- | -- | 100 | 100 |
| | TSA | 2 | 4 | 4 | -- | -- | 100 | |
| Serratia marcescens | MAC | 4 | 8 | 8 | -- | -- | 100 | 100 |
| | TSA | 4 | 8 | 8 | -- | -- | 100 | |
| Stenotrophomonas maltophilia | MAC | 2 | 4 | 4 | -- | -- | 100 | 100 |
| | TSA | 2 | 4 | 4 | -- | -- | 100 | |
| Vibrio parahaemolyticus | TSA | 1 | 2 | 2 | -- | -- | 100 | 100 |
| | Total (%) | 156 | 312 (100) | 296 (94.9) | 10 (3.2) | 6 (1.9) | | |

No ID: No identification; BG: Bordet Gengou Agar + 15% Sheep Blood; CHO: Chocolate Agar; MAC: MacConkey Agar; TSA: Trypticase Soy Agar + 5% Sheep Blood
1 Agreement with Expected Organism Identity with High Confidence Log(Score).
2 Includes reports of "No Identification" and "No peaks".
3 2 colonies of each strain were designated for picking by the Colibri System and each was used to prepare a separate target spot.

Table 18. Summary of results from representative Gram-positive bacteria using the Copan Colibri System in conjunction with the Bruker MALDI Biotyper CA using 96-spot targets.

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| Colibrí System with MALDI Biotyper CA and 96-spot target: Gram-Positive Bacteria | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Expected Organism Identity | Culture Medium | Strains Tested | Colonies Picked3 | MALDI Biotyper CA Identification Result | | | % Agreement1 | |
| | | | | Confidence | | No ID2 | Culture Medium | Species/Group |
| | | | | High | Low | | | |
| Enterococcus faecalis | TSA | 6 | 12 | 9 | -- | 3 | 75.0 | 87.5 |
| | CNA | 6 | 12 | 12 | -- | -- | 100 | |
| Enterococcus faecium | TSA | 6 | 12 | 9 | 1 | 2 | 75.0 | 83.3 |
| | CNA | 6 | 12 | 11 | 1 | -- | 91.7 | |
| Listeria monocytogenes | TSA | 2 | 4 | 4 | -- | -- | 100 | 100 |
| Staphylococcus aureus | TSA | 6 | 12 | 10 | 1 | 1 | 83.3 | 91.7 |
| | CNA | 6 | 12 | 12 | -- | -- | 100 | |
| Staphylococcus epidermidis | TSA | 6 | 12 | 5 | 5 | 2 | 41.7 | 37.5 |
| | CNA | 6 | 12 | 4 | 6 | 2 | 33.3 | |
| Staphylococcus saprophyticus | TSA | 4 | 8 | 4 | 1 | 3 | 50.0 | 56.3 |
| | CNA | 4 | 8 | 5 | 1 | 2 | 62.5 | |
| Streptococcus agalactiae | TSA | 6 | 12 | 4 | 6 | 2 | 33.3 | 29.2 |
| | CNA | 6 | 12 | 3 | 4 | 5 | 25.0 | |
| Streptococcus pyogenes | TSA | 4 | 8 | 7 | 1 | -- | 87.5 | 81.3 |
| | CNA | 4 | 8 | 6 | 2 | -- | 75.0 | |
| | Total (%) | 78 | 156 (100) | 105 (67.3) | 29 (18.6) | 22 (14.1) | | |

No ID: No identification; CNA: Columbia Agar + 5% Sheep Blood with colistin and nalidixic acid (CNA); TSA: Trypticase Soy Agar + 5% Sheep Blood
1 Agreement with Expected Organism Identity with High Confidence Log(Score).
2 Includes reports of "No Identification" and "No peaks".
3 2 colonies of each strain were designated for picking by the Colibri System and each was used to prepare a separate target spot.

Table 19. Comparison of Bruker MALDI Biotyper CA results from targets prepared on the Copan Colibri System or manually.

| Organism | Target Type | Colonies Picked | MALDI Biotyper CA Identification Result | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | | High Confidence | | Low Confidence | | No Identification | |
| | | | Colibrí | Manual | Colibrí | Manual | Colibrí | Manual |
| Gram-Negative | 48-spot | 284 | 281 (98.9) | 281 (98.9) | 3 (1.1) | 2 (0.7) | 0 (0.0) | 1 (0.3) |
| | 96-spot | 312 | 296 (94.9) | 289 (92.6) | 10 (3.2) | 12 (3.8) | 6 (1.9) | 11 (3.5) |
| Gram-Positive | 48-spot | 156 | 128 (82.0) | 148 (94.9) | 22 (14.1) | 8 (5.1) | 6 (3.8) | 0 (0.0) |
| | 96-spot | 156 | 105 (67.3) | 123 (78.8) | 29 (18.6) | 23 (14.7) | 22 (14.1) | 10 (6.4) |
| Total | | 908 | 810 (89.2) | 841 (92.6) | 64 (7.0) | 50 (5.5) | 34 (3.7) | 22 (2.4) |

# Accuracy of Bacterial Identification (Specificity)

To verify the specificity of the Copan Colibri System and that automated sample preparation does not affect MALDI-TOF MS identification results, targets were prepared using colonies of representative "off-panel" species (10 isolates per analyzer, 2 spots per isolate) that are not

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included in the clinically validated databases of the VITEK MS and MALDI Biotyper CA systems. Each of the species tested produced the expected "No identification" results with both mass spectrometers, demonstrating that automated sample preparation did not adversely affect the specificity of MALDI-TOF MS identification. These results are acceptable.

## Positional Accuracy

Studies were conducted to evaluate the ability of the Copan Colibri System to prepare target spots for MALDI-TOF MS analysis at each location on the targets for both the bioMérieux VITEK MS and Bruker MALDI Biotyper CA. Testing was done with isolated colonies of *E. coli* and *S. aureus* grown on Trypticase Soy Agar containing 5% sheep blood. Each of three Colibri Systems was used to prepare spots at each location on three separate targets for each analyzer. Both 48-spot reusable and 96-spot disposable targets were included in the study with the Bruker MALDI Biotyper CA. The results obtained were analyzed in terms of percent agreement with the expected organism identity at each location (Table 20). No positional effects were observed, although lower agreement was observed with *S. aureus* than with *E. coli*, which is consistent with other studies with the Colibri System in which agreement with the expected organism identity was lower for Gram-positive than Gram-negative species. These results are acceptable.

Table 20. Summary of results from evaluation of Copan Colibri System positional accuracy.

| MALDI-TOF MS Analyzer | Target | Expected Identity | Spots Prepared | Agreement with Expected Organism Identity (%) | | No Identification |
| --- | --- | --- | --- | --- | --- | --- |
| | | | | High/Good Confidence | Low Confidence/Low Discrimination | |
| VITEK MS | 48-spot | E. coli | 432 | 432 (100) | 0 (0) | 0 (0) |
| | | S. aureus | 432 | 431 (99.8) | 0 (0) | 1 (0.2) |
| MALDI Biotyper CA | 48-spot | E. coli | 432 | 431 (99.8) | 1 (0.2) | 0 (0) |
| | | S. aureus | 432 | 418 (96.8) | 14 (3.2) | 0 (0) |
| | 96-spot | E. coli | 846 | 845 (99.9) | 1 (0.1) | 0 (0) |
| | | S. aureus | 846 | 810 (95.7) | 34 (4.0) | 2 (0.2) |

## 5. Carry-Over:

To evaluate the potential for cross-contamination between target spots due to positional inaccuracy of the Copan Colibri System, testing was performed by spotting alternating patterns of six "on-panel" and six "off-panel" species onto targets for each of the two mass spectrometry instruments. "On panel" bacterial species, i.e., those represented in the FDA-cleared MS reference databases, were obtained from ATCC, and "off-panel" species that were not included in the databases of the VITEK MS or MALDI Biotyper CA systems were clinical isolates from bioMérieux identified via 16S rRNA sequencing and Copan identified via the MALDI Biotyper MBT Compass (RUO). The targets were analyzed and the reported identity for each spot was compared to the expected value. Results are summarized in Tables 21 and 22 for the bioMérieux VITEK MS and Bruker MALDI Biotyper CA, respectively. There was no evidence of cross-contamination with targets prepared for either mass spectrometry system, and no incorrect identification results were obtained. However, with *E. faecalis* and *S. agalactiae* on targets for the Bruker MALDI Biotyper CA, fewer High Confidence Log(Score) values were observed than for other bacterial species, leading to lower positive percent agreement. This is consistent with observations in other Analytical Studies using the Colibri System in conjunction with the Bruker MALDI Biotyper CA to identify Gram-positive organisms and is mitigated by the requirement for additional testing

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that is noted in the device labeling. The results from the Carryover/Cross-Contamination Study were therefore determined to be acceptable.

Table 21. Results of the cross-contamination study for the Copan Colibri System with the bioMérieux VITEK MS.

| Colibri Spotting Mode | “On-panel” Species | Positive Agreement (%)1 | “Off-panel” Species | Negative Agreement (%)2 |
| --- | --- | --- | --- | --- |
| Duplicate | Escherichia coli | 24/24(100) | Burkholderia thailandensis | 24/24(100) |
| | Klebsiella pneumoniae | 24/24(100) | Pseudocitrobacter faecalis | 24/24(100) |
| | Pseudomonas aeruginosa | 24/24(100) | Acidovorax delafieldii | 24/24(100) |
| | Enterococcus faecalis | 22/22(100) | Rothia amarae | 22/22(100) |
| | Staphylococcus aureus | 24/24(100) | Leuconostoc carnosum | 24/24(100) |
| | Streptococcus agalactiae | 23/243(95.8) | Aneurinibacillus migulanus | 24/24(100) |
| | All “On-panel” Species | 141/142(99.3) | All “Off-panel” Species | 142/142(100) |
| Single | Escherichia coli | 24/24(100) | Burkholderia thailandensis | 24/24(100) |
| | Klebsiella pneumoniae | 24/24(100) | Pseudocitrobacter faecalis | 24/24(100) |
| | Pseudomonas aeruginosa | 24/24(100) | Acidovorax delafieldii | 24/24(100) |
| | Enterococcus faecalis | 24/24(100) | Rothia amarae | 24/24(100) |
| | Staphylococcus aureus | 24/24(100) | Leuconostoc carnosum | 24/24(100) |
| | Streptococcus agalactiae | 23/243(95.8) | Aneurinibacillus migulanus | 24/24(100) |
| | All “On-panel” Species | 143/144(99.3) | All “Off-panel” Species | 144/144(100) |

1 Expected organism identity with Good Confidence.
2 Expected result $=$ "No identification".
3 1 colony gave a result of "No identification".

Table 22. Results of the cross-contamination study for the Copan Colibri System with the Bruker MALDI Biotyper CA.

| Target (Spotting Mode) | “On-panel” Species | Positive Agreement (%)1 | “Off-panel” Species | Negative Agreement (%)2 |
| --- | --- | --- | --- | --- |
| 48-spot Reusable (Duplicate) | Acinetobacter baumannii | 23/24 (95.8) | Novosphingobium capsulatum | 24/24 (100) |
| | Escherichia coli | 56/56 (100) | Cedecea neteri | 56/56 (100) |
| | Klebsiella pneumoniae | 24/24 (100) | Gallibacterium anatis | 24/24 (100) |
| | Enterococcus faecalis | 19/243 (79.2) | Bacillus infantis | 24/24 (100) |
| | Staphylococcus aureus | 48/48 (100) | Bacillus flexus | 48/48 (10…