Colibrí System

K220546 · Copan Wasp Srl · LON · Oct 5, 2022 · Microbiology

Device Facts

• Record ID: K220546
• Device Name: Colibrí System
• Applicant: Copan Wasp Srl
• Product Code: LON · Microbiology
• Decision Date: Oct 5, 2022
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.1645
• Device Class: Class 2

Intended 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 and with the bioMérieux VITEK 2 Antimicrobial Susceptibility Testing (AST) system for qualitative testing 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 for bacterial identification and microbial suspension at known concentration for antimicrobial susceptibility testing and purity assessment. 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 analyzers. Bacterial suspensions for AST and purity plates are identified by barcode label. 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 the identification or processing of yeast species, molds, Nocardia, or mycobacteria.

Device Story

The Colibrí System is a semi-automated pre-analytical processor for clinical microbiology laboratories. It consists of a Vision System and a Preparation Station. Operators use a graphical interface to select isolated bacterial colonies from digital images of culture plates. The system uses a robotic pipetting arm to pick the designated colonies and either spot them onto MALDI-TOF MS target slides or suspend them in saline for AST. An onboard nephelometer measures suspension turbidity to ensure correct concentration for VITEK 2 AST cards. The system labels secondary tubes and purity plates for traceability and communicates sample identity to downstream analyzers. By automating colony picking and suspension preparation, the device reduces manual labor, cross-contamination risks, and variability in inoculum density, ultimately supporting accurate bacterial identification and susceptibility testing.

Clinical Evidence

No clinical trials were performed. Evidence consists of analytical bench testing, including nephelometer calibration verification (100% accuracy), pipettor trueness/precision, and purity plate assessments (2,364 plates, 100% monomicrobial). AST challenge testing compared Colibrí-prepared suspensions to manual preparation across 236 isolates (Enterobacterales, Staphylococcus, Streptococcus, Enterococcus, non-fermenters). Results showed >99.9% Essential Agreement (EA) and 99.3% Category Agreement (CA). Reproducibility studies confirmed consistency across instruments, operators, and days.

Technological Characteristics

System comprises Colibrí Vision System and Preparation Station. Uses sterile pipette tips, primary/secondary tubes, and spreader. Sensing: onboard nephelometer for turbidity (McFarland). Connectivity: electronic communication to MALDI-TOF MS analyzers. Software: rule-based image processing for colony selection and automated pipetting control. Sterilization: onboard incinerator for spreader. Dimensions/form factor: benchtop instrument.

Indications for Use

Indicated for trained healthcare professionals in clinical laboratories to aid in the diagnosis of bacterial infections by semi-automated processing of isolated gram-negative and gram-positive bacterial colonies grown on solid culture media. Used for MALDI-TOF MS identification and AST suspension preparation. Not validated for yeast, molds, Nocardia, or mycobacteria.

Regulatory Classification

Identification

A fully automated short-term incubation cycle antimicrobial susceptibility system is a device that incorporates concentrations of antimicrobial agents into a system for the purpose of determining in vitro susceptibility of bacterial pathogens isolated from clinical specimens. Test results obtained from short-term (less than 16 hours) incubation are used to determine the antimicrobial agent of choice to treat bacterial diseases.

Special Controls

*Classification.* Class II (special controls). The special control for this device is FDA's guidance document entitled “Class II Special Controls Guidance Document: Antimicrobial Susceptibility Test (AST) Systems; Guidance for Industry and FDA.”

Predicate Devices

• Vitek 2 Systems (pc) 5.02 Software (K103752)

Submission Summary (Full Text)

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October 5, 2022

Copan WASP S.r.l. Chiara Congiu Regulatory Affairs Via A. Grandi, 32 Brescia, Brescia 25125 Italy

## Re: K220546

Trade/Device Name: Colibrí System Regulation Number: 21 CFR 866.1645 Regulation Name: Fully Automated Short-Term Incubation Cycle Antimicrobial Susceptibility System Regulatory Class: Class II Product Code: LON, QQV, QBN Dated: February 18, 2022 Received: February 25, 2022

## Dear Chiara Congiu:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part

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801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email ([e-mail]) or phone [phone] or 301-796-7100).

Sincerely,

For: Uwe Scherf, Ph.D. Director Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Ouality Center for Devices and Radiological Health

Enclosure

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## Indications for Use

510(k) Number (if known) K220546

Device Name Colibrí System

#### Indications for Use (Describe)

The Collbri™ System is an in vitro diagnostic device comprised of the Collbri™ Preparation Station for use with the bioMérieux VITEK® MS or Bruker MALDI Biotyper® CA mass spectrometry systems for qualitative identification and with the bioMérieux VITEK® 2 Antimicrobial Susceptibility Testing (AST) system for qualitative testing of isolated colonies of gram-negative and gram-positive bacterial species grown on solid culture media. The Collbri™ 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/Jonization-Time of Flight Mass Spectrometry) target slides for bacterial identification and microbial suspension at known concentration for Antimicrobial Susceptibility Testing and purity assessment.

The Colibri™ software records the identity of each sample and its position on the target slide and communicates this information electronically to the MALDI-TOF MS analyzers.

Bacterial suspensions for AST and purity plates are identified by barcode label.

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

The Collbri™ System has not been validated for use in the identification or processing of yeast species, Mocardia, or mycobacteria.

| Type of Use (Select one or both, as applicable) |
|------------------------------------------------------------------------------------|
| <span style="font-size:10px">☑</span> Prescription Use (Part 21 CFR 801 Subpart D) |
| <span style="font-size:10px">☐</span> Over-The-Counter Use (21 CFR 801 Subpart C) |

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#### I. Submitter

| Applicant Name: | Copan WASP Srl<br>Via A. Grandi 32<br>25125 Brescia, Italy<br>[phone]<br>[e-mail] |
|------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------|
| Contact Person | Chiara Congiu<br>Copan WASP Srl<br>Via A. Grandi 32<br>25125 Brescia, Italy<br>[phone]<br>[e-mail] |
| Establishment Registration Number: | 3009288740 |
| Date Prepared: | February 18, 2022 |

#### II. Device Name

| Proprietary Name | Colibrí System |
|---------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Common/Usual Name | Colibrí System |
| Classification Name | Fully automated short-term incubation cycle antimicrobial susceptibility system (21 CFR 866. 1645)<br>Clinical mass spectrometry microorganism identification and differentiation system (21 CFR 866.3378) [Cleared, K193138] |
| Device Class | II |
| Product Code | LON |
| Panel | QQV, QBN [Cleared, K193138]<br>Microbiology |

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#### III. Legally Marketed Predicate Device

| Device Name | VITEK 2 Gram Negative Imipenem & VITEK 2<br>Systems (PC) 5.02 Software |
|---------------|------------------------------------------------------------------------|
| 510(K) Number | K103752 |

No reference Devices were used in this submission.

#### IV. Device Description

The Copan Colibrí System is designed to be used as an accessory of the downstream MALDI-TOF MS and antimicrobial susceptibility testing (AST) analyzers automating various manual steps in the workflow for the preparation of samples for the identification of isolated colonies and for AST of isolated colonies of gram-negative and gram-positive bacterial species grown on solid culture media.

The Colibrí System automates the preparation of MALDI target slides for the bioMérieux VITEK MS or the Bruker MALDI Biotyper CA System that are used in clinical laboratories for identification (ID) of organisms grown on plated media by Matrix-Assisted Laser Desorption/Jonization Time-of Flight Mass Spectrometry (MALDI-TOF MS). The Colibri System automates the preparation of microbial suspensions at known concentration for bioMérieux VITEK 2 System that is used in clinical laboratories for AST analyses. Moreover, the Colibri System is used for Purity Plates preparation for purity assessments.

The Colibrí System comprises the Colibrí Vision System and Colibrí Preparation Station hardware modules and pipette tips, Primary Tubes, Spreader and nephelometer Verification Kit as consumables. After appropriate plate incubation, the operator using the graphical User Interface (Image Reading Interface) chooses the plates exhibiting adequate growth and selects the isolated colonies to be processed assigning the automatic ID or AST tasks. By using the Colibrí Vision System, specific colonies to be picked are designated by the operator on a digital plate. The Operator manually loads the plates in the Colibri Preparation Station where colonies are automatically picked, spotted on the target slide and overlayed with the matrix or suspended into the dedicated solution for the preparation of the microbial suspension for AST purposes (Secondary Tube).

When used in conjunction with the bioMérieux VITEK MS, the Colibrí System can prepare the 48spot target slides by performing the direct spotting of colonies. The calibrator used for quality control is manually applied by the operator at the end of the automated colony spotting. When used in conjunction with the Bruker MALDI Biotyper CA System, the Colibrí System can prepare either reusable 48-spot or disposable 96-spot targets by performing the Direct Transfer Sample Procedure. The BTS used for quality control is manually applied by the operator at the automated colony spotting.

When used in conjunction with the bioMérieux VITEK 2, the Colibrí System can prepare the microbial suspension at the proper concentration by direct colony suspension method. The onboard nephelometer allows the preparation of Secondary Tubes (AST suspensions) at the correct concentration and the Colibrí Spreader is used for Purity Plates preparation.

Copan WASP S.r.l., Traditional 510(k)- Colibrí System

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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 analyzers.

The traceability of prepared Secondary Tube and Purity Plates is maintained by dedicated labels applications.

Colibri System requires four different calibrations, one on the nephelometer, three on the cameras. None of these calibration activities require user intervention if not in terms of periodical cleaning of the mechanical component as described in the dedicated section of the User Manual. The Set-up calibration of nephelometer and camera units positioned on the Colibrí Vision System and on the Colibrí Preparation Station are performed during the device initial setup. Auto-calibration is performed at the end of the initial set-up and periodically during the preventive maintenance to check that, in the Collbrí Preparation all the mechanical references can be found inside the positioning tolerances, that the I/Os are responsive. Run-time calibration is performed during the normal usage to automatically check the proper functioning of the Colibrí Vision System and the Colibrí Preparation Station.

Colibrí System requires a daily nephelometer verification to check the proper reading of suspensions at different turbidity values.

#### V. 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 and with the bioMérieux VITEK 2 Antimicrobial Susceptibility Testing (AST) system for qualitative testing 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 a pipetting system to prepare operator and uses MALDI-TOF MS (Matrix-Laser Desorption/Ionization- Time Of Flight Mass Spectrometry) target slides for Assisted bacterial identification and microbial suspension at known concentration for Antimicrobial Susceptibility Testing and purity assessment.

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.

Bacterial suspensions for AST and Purity Plates are identified by barcode label.

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 the identification or processing of yeast species, molds, Nocardia, or mycobacteria.

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#### VI. Comparison to Predicate

Colibrí System is designed to automatize the standard manual workflow for the preparation of microbial suspension for AST via direct colony suspension method and purity assessment decreasing the risk of cross-contamination among colonies grown on the culture plate, the risk of scratching from the media plate surface and the risk to use AST suspensions at improper concentration. Specifically, the Colibrí Vision System aids the operator in selecting single, well-isolated colonies. The Colibrí Preparation Station allows the automatic picking of the preselected colonies and their suspension into the saline solution of the Primary Tube. The Primary Tube turbidity is checked by the on-board nephelometer to assure it is in the proper working range, allowing the preparation of a Secondary Tube at a precise concentration. The Colibri Preparation Station labels the Secondary Tube and the Purity Plate, optionally prepared, for traceability.

With reference to the sample preparation workflow for AST testing, comparison with the Predicate Device is provided in the following tables:

| Similarities | | |
|---------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Item | New Device | Predicate Device |
| Device Name<br>(K number) | Colibrí System<br>(K220546) | VITEK 2 Gram Negative Imipenem &<br>VITEK 2 Systems (PC) 5.02 Software<br>(K103752) |
| Device Classification | Class II (special controls) | Class II (special controls) |
| Regulation Number | 21 CFR 866.1645 Fully automated short-term incubation cycle antimicrobial susceptibility system | 21 CFR 866.1645 Fully automated short-term incubation cycle antimicrobial susceptibility system |
| Product Code | LON System, Test, Automated, Antimicrobial Susceptibility, Short Incubation | LON System, Test, Automated, Antimicrobial Susceptibility, Short Incubation |
| 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 and with the bioMérieux VITEK 2 Antimicrobial Susceptibility Testing (AST) system for qualitative testing 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 and microbial suspension at known concentration for Antimicrobial Susceptibility Testing and purity assessment.<br>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.<br>Bacterial suspensions for AST and Purity Plates are identified by barcode label.<br>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 | VITEK 2 GN Imipenem is designed for antimicrobial susceptibility testing of Gram-Negative bacilli. VITEK 2 GN Imipenem is a quantitative test intended for use with the VITEK 2 and VITEK 2 Compact Systems using VITEK 2 Systems 5.02 Software as a laboratory aid in the determination of in vitro susceptibility to antimicrobial agents. Imipenem has been shown to be active both in vitro and in clinical infections against most strains of the following microorganisms according to the FDA label for the antimicrobial.<br>Active in vitro and in clinical infections:<br>Acinetobacter spp.<br>Citrobacter spp.<br>Enterobacter aerogenes<br>Escherichia coli<br>Klebsiella spp.<br>Active in vitro but clinical significance unknown:<br>Providencia stuartii<br>The VITEK 2 Antimicrobial Susceptibility Test (AST) is intended to be used with the VITEK 2 |
| | | Morganella morganii<br>Proteus vulgaris<br>Providencia rettgeri<br>Pseudomonas aeruginosa<br>Serratia marcescens |

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| Similarities | | |
|----------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Item | New Device | Predicate Device |
| Device Name<br>(K number) | Colibrí System<br>(K220546) | VITEK 2 Gram Negative Imipenem &<br>VITEK 2 Systems (PC) 5.02 Software<br>(K103752) |
| | diagnosis of bacterial infections.<br>The Colibrí System has not been validated for<br>use in the identification or processing of yeast<br>species, molds, Nocardia or mycobacteria. | System for the automated quantitative or<br>qualitative susceptibility testing of isolated<br>colonies for the most clinically significant<br>aerobic Gram-Negative bacilli, <i>Staphylococcus</i><br>spp., <i>Enterococcus</i> spp., <i>Streptococcus</i><br>agalactiae, <i>S. pneumoniae</i> and clinically<br>significant yeast. The VITEK 2 Systems (PC)<br>5.02 Software is intended for use with VITEK 2<br>and VITEK 2 Compact Systems. |
| Method of testing | Direct testing from isolated colonies. | Direct testing from isolated colonies. |
| Sample/Media Type | Isolated bacterial colonies from any patient<br>source. | Isolated bacterial colonies from any patient<br>source. |
| | Acceptable media:<br>1. Trypticase soy agar with 5% sheep blood<br>2. MacConkey agar<br>3. Columbia blood agar with 5% sheep blood<br>as whole plate or bi-plate | Acceptable media:<br>1. Trypticase soy agar with 5% sheep blood<br>2. MacConkey agar<br>3. Columbia blood agar with 5% sheep blood |
| Solution for Suspension<br>Preparation | Aqueous 0.45% NaCl Saline Solution (pH 4.5 to<br>7.0)<br>3mL volume in the Secondary Tube | Aqueous 0.45% NaCl Saline Solution (pH 4.5 to<br>7.0)<br>3mL volume in the Secondary Tube |
| Inoculum density check | The accuracy of the inoculum preparation is<br>verified by an on-boarding nephelometer. | The accuracy of the inoculum preparation is<br>verified by a nephelometer. |
| Quality control | Suspension of reference strains to be used as<br>quality control should be prepared manually<br>according to the instruction for use of the used<br>VITEK 2 card. | Suspension of reference strains to be used as<br>quality control should be prepared manually<br>according to the instruction for use of the used<br>VITEK 2 card. |
| AST results<br>interpretation | MIC and categorization of results are provided<br>by VITEK 2 | MIC and categorization of results are provided<br>by VITEK 2 |

| Differences | | |
|-----------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Item | New Device | Predicate Device |
| Device Name<br>(K number) | Colibrí System<br>(K220546) | VITEK 2 Gram Negative Imipenem &<br>VITEK 2 Systems (PC) 5.02 Software<br>(K103752) |
| Colony Selection | The colony to be picked is selected by an<br>operator on a digital plate using the Graphical<br>User Interface of a Vision System. | The colony to be picked be picked is manually<br>selected by an operator on a real plate through<br>the visual inspection. |
| Media Type | Colibrí System is not validated for ChromID CPS | Acceptable media:<br>1. ChromID CPS |
| Method of Colony<br>Picking | Colibrí System has been validated for automatic<br>picking of colonies using a sterile pipette tip. | The colonies to be picked are manually<br>transferred using a sterile stick or swab. |
| Sample Traceability | On each Secondary Tube prepared by the Colibrí<br>System, a barcode label is applied including<br>following data: the sample identification, the<br>hour of the preparation and the Gram<br>classification associated to the processed isolate.<br>Label data are used for sample traceability for<br>further processing on the VITEK 2. | The sample identification is recorded directly<br>in the Cassette Docking Station software<br>manually or scanning the barcode of the<br>culture media plate from which the colonies<br>were collected during the preparation of the<br>microbial suspension. |
| Method of AST<br>suspension preparation | Using a pipetting system, a predefined number<br>of morphologically similar colonies are<br>transferred into Primary Tube containing saline<br>solution (0.45% NaCl Saline Solution pH 4.5 to<br>7.0). A homogenous heavy suspension of<br>organisms is prepared and checked by using on- | Using a sterile stick or swab, a sufficient<br>number of morphologically similar colonies<br>are transferred to a saline tube (0.45% NaCl,<br>Saline Solution pH 4.5 to 7.0). A homogenous<br>suspension with a density equivalent to the 0.5<br>McFarland is prepared and checked with the |

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| | Differences | | |
|---------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--|
| Item | New Device | Predicate Device | |
| Device Name<br>(K number) | Colibrí System<br>(K220546) | VITEK 2 Gram Negative Imipenem &<br>VITEK 2 Systems (PC) 5.02 Software<br>(K103752) | |
| | board Colibrí nephelometer. In the Secondary<br>Tube containing 3.0mL of the same saline<br>solution, a variable aliquot of the heavy<br>suspension is automatically transferred to obtain<br>the final microbial concentration according to<br>IVD package insert indications. The suspensions<br>prepared by Colibrí System must be tested in<br>MANUAL MODE on the VITEK 2. | nephelometer. In a second tube containing<br>3.0mL of saline, a predetermined aliquot of 0.5<br>McFarland is transferred according to IVD<br>package insert indications (MANUAL<br>MODE).<br>Alternatively, the 0.5 McFarland suspension<br>is loaded on the VITEK 2 that automatically<br>prepares the Secondary Tube at proper<br>concentration (AUTO DILUTION MODE). | |

These differences do not affect substantial equivalence of Colibrí System and the Predicate Device. Both Systems are intended for the AST of microorganisms cultured from human specimens

#### VII. Performance Data

The following performance data were provided in support of the substantial equivalence determination.

### Analytical Studies

The Analytical studies performed with the Colibri System support its use for the preparation of microbial suspension used in conjunction with the bioMérieux VITEK 2 AST analyzer. The Analytical studies demonstrated that the Device can automatically prepare the microbial suspensions at appropriate concentrations, starting from gram-negative and ram-positive bacterial colonies grown on solid culture media, which can be used to hydrate VITEK 2 cards for the determination of susceptibility of organisms to certain drugs. The used methodology (direct colony suspension) and claimed prerequisites for sample preparation are in line with the IVD analyzer manufacturer IFU.

#### Nephelometer Calibration Verification

To verify the accuracy of the onboard Colibrí System nephelometer in preparing microbial suspensions at specific concentrations within the calibration range, isolated colonies of E. coli (ATCC 25922) grown on non-selective medium were used to manually prepare suspensions at determined concentrations (0.25, 0.5, 1.0, 2.0, 3.0 McFarland), representing the calibration points. For each concentration, 20 suspensions were prepared from three operators in rotation, and the process was repeated on 3 Colibrí Systems calibrated with 3 different lots of suspensions at known concentrations. For each suspension, ten-fold dilutions were prepared and plated in triplicate; to perform viable cell count to calculate the initial tube concentration.

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A total of 300 suspensions were prepared: overall, 100% of suspensions contained the correct concentration of bacteria considering that a 0.5 McFarland suspension of E. coli has a nominal microbial content of 1-2 × 10° CFU/mL+. The study demonstrated acceptable accuracy.

### Pipettor Trueness and Precision

The accuracy and reproducibility of the on-board Colibrí System pipettor was determined gravimetrically. Appropriate vessels were weighted before and after the dispensations of four volumes (50uL, 100 uL, 500uL, 900uL) representing the 5%, 10%, 50% and 90% of the nominal volume of the tip used for the AST preparation. Three Colibrí System pipettors were included in the examination: for each volume, 10 measurements were performed using saline solution (aqueous 0.45%, pH 4.5 to 7.0) and the trueness and reproducibility were calculated. As expected, trueness and reproducibility vary according to the volume under testing but always within the acceptance criteria.

### E. coli Suspensions Preparation Verification Study

To assess the ability of the Colibri System to prepare and manage Primary Tubes at various concentrations, isolated colonies of the E. coli ATCC® 25922 were used to automatically prepare Primary Tubes at various turbidities.

A variable number of colonies was selected on the plates images to create different suspensions at increasing turbidity values. Three Colibrí Systems run by three different operators were included in the test.

All the Primary Tubes were correctly managed by Colibrí System according to the turbidity value.

100% suspensions over the entire working range contained the expected number of colonies, estimated considering that the 0.5 McFarland has a nominal content of 1-2 × 108 CFU/mL for E. coli'.

### Validation of Colony Picking and Preparation of Microbial Suspensions for AST

The accuracy of colony picking and preparation of the microbial suspension was demonstrated by purity check and bacterial concentration determination of the Primary Tubes.

Three Colibrí Systems were used to prepare Primary Tubes and the respective subculture (Purity Plate) from isolated colonies of 6 bacterial species (3 Gram-Negative and 3 Gram-Positive) grown in 2 polymicrobial mixtures on different types of culture medium in whole and biplates at various incubation time. To confirm nephelometer accuracy, bacterial concentration of the Primary Tube was determined by viable cell count and compared to the theorical concentration, estimated considering that the 0.5 McFarland has a nominal content of 1-2 × 108 CFU/mL for E. coli¹.

¹ CLSI guideline M07. 11th ed Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically. Wayne, PA: Clinical and Laboratory Standards Institute, 2018.

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100% of colonies designated by the operator were picked correctly by the Colibrí System both on whole plates and bi-plates and 100% of Purity Plates showed no evidence of microbial contamination, demonstrating that the Colibrí System accurately picks microbial colonies, without contamination from other microorganisms grown on the same culture plate.

The percentage of prepared suspension with microbial concentration within the acceptable limits was 99.2% and, for each instrument, the result was always >98%. The percentage suspension within acceptable limits has been compared between each instrument with y-test, resulting in no statistically significant difference among the instruments. The results provided evidence that the microbial concentration is accurately measured by the Colibri nephelometer through its turbidity.

## AST Challenge Test

The accuracy of MICs obtained by bioMérieux VITEK 2 using Colibrí System for microbial suspensions preparation was evaluated using representative isolates of different species of Enterobacterales (n=62), Staphylococcus (n=16), Streptococcus (n=30), Enterococcus (n=16) and non-fermenters (n=32). The strains included in this study were both susceptible and resistant strains, exhibiting a range of on-scale MIC values toward at least 4 antibiotics representative for the major classes of drugs. Each strain was grown on different agar media (Trypticase Soy Agar + 5% Sheep Blood, MacConkey Agar and Columbia agar+5% sheep blood) at specific incubation times and then processed on three Colibrí System in comparison with the manual preparation.

Microbial suspensions were prepared by Colibrí Preparation Station and processed by the bioMérieux VITEK 2 using the appropriate antibiotic panel. The MICs obtained by bioMérieux VITEK 2 using Colibri System were compared to the MICs obtained by bioMérieux VITEK 2 using manual sample preparation and the SIR category was reported according to the FDA- Recognized Antimicrobial Susceptibility Test Interpretive Criteria. Essential Agreement (EA) of the MICs and Category Agreement (CA) were calculated. The discrepant SIR results were categorized as Very Major Category Error, Major Category Error and Minor Category Error.

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| Agent | Organism group | Total tested | # EA | % EA | Total Evaluable | # EA of Evaluable | % EA of Evaluable | Total cat. | # CA | % CA | # S | # R | # vmj | # maj | # min |
|---------------------------------|------------------|--------------|------|-------|-----------------|-------------------|-------------------|------------|------|-------|-----|-----|-------|-------|-------|
| Amikacin | Enterobacterales | 186 | 186 | 100% | 81 | 81 | 100% | 186 | 182 | 97.8% | 141 | 39 | 0 | 0 | 4 |
| Amikacin | Non-fermenters | 81 | 81 | 100% | 39 | 39 | 100% | 81 | 79 | 97.5% | 66 | 6 | 0 | 0 | 2 |
| Ampicillin | Enterococcus | 48 | 48 | 100% | 4 | 4 | 100% | 48 | 48 | 100% | 36 | 12 | 0 | 0 | 0 |
| Ampicillin | Streptococcus | 90 | 90 | 100% | 0 | 0 | N/A | 90 | 90 | 100% | 90 | 0 | 0 | 0 | 0 |
| Ampicillin / Sulbactam | Enterobacterales | 111 | 111 | 100% | 9 | 9 | 100% | 111 | 111 | 100% | 21 | 90 | 0 | 0 | 0 |
| Ampicillin / Sulbactam | Non-fermenters | 24 | 24 | 100% | 5 | 5 | 100% | 24 | 23 | 95.8% | 15 | 6 | 0 | 0 | 1 |
| Aztreonam | Enterobacterales | 186 | 185 | 99.5% | 54 | 54 | 100% | 186 | 185 | 99.5% | 57 | 123 | 0 | 0 | 1 |
| Cefepime | Enterobacterales | 186 | 186 | 100% | 58 | 58 | 100% | 186 | 185 | 99.5% | 93 | 54 | 0 | 0 | 1 |
| Cefepime | Non-fermenters | 81 | 81 | 100% | 50 | 50 | 100% | 81 | 80 | 98.8% | 48 | 33 | 0 | 1 | 0 |
| Cefotaxime | Streptococcus | 90 | 90 | 100% | 0 | 0 | N/A | 90 | 90 | 100% | 90 | 0 | 0 | 0 | 0 |
| Cefoxitin | Enterobacterales | 186 | 186 | 100% | 42 | 42 | 100% | 186 | 184 | 98.9% | 36 | 132 | 0 | 0 | 2 |
| Ceftazidime | Enterobacterales | 180 | 180 | 100% | 43 | 43 | 100% | 180 | 179 | 99.4% | 33 | 132 | 0 | 0 | 1 |
| Ceftazidime | Non-fermenters | 105 | 105 | 100% | 68 | 68 | 100% | 105 | 105 | 100% | 57 | 36 | 0 | 0 | 0 |
| Ceftriaxone | Enterobacterales | 186 | 185 | 99.5% | 31 | 30 | 96.8% | 186 | 185 | 99.5% | 36 | 144 | 0 | 0 | 1 |
| Ceftriaxone | Streptococcus | 90 | 90 | 100% | 0 | 0 | N/A | 90 | 90 | 100% | 90 | 0 | 0 | 0 | 0 |
| Ciprofloxacin | Non-fermenters | 81 | 81 | 100% | 28 | 28 | 100% | 81 | 80 | 98.8% | 51 | 18 | 0 | 0 | 1 |
| Ciprofloxacin | Staphylococcus | 48 | 48 | 100% | 6 | 6 | 100% | 48 | 48 | 100% | 33 | 12 | 0 | 0 | 0 |
| Clindamycin | Enterococcus | 48 | 48 | 100% | 30 | 30 | 100% | 48 | 47 | 97.9% | 27 | 12 | 0 | 0 | 1 |
| Clindamycin | Streptococcus | 90 | 90 | 100% | 2 | 2 | 100% | 90 | 89 | 98.9% | 54 | 33 | 0 | 0 | 1 |
| Ertapenem | Enterobacterales | 186 | 186 | 100% | 22 | 22 | 100% | 186 | 184 | 98.9% | 102 | 81 | 0 | 0 | 2 |
| Erythromycin | Staphylococcus | 48 | 48 | 100% | 9 | 9 | 100% | 48 | 47 | 97.9% | 21 | 24 | 0 | 0 | 1 |
| Erythromycin | Enterococcus | 48 | 48 | 100% | 15 | 15 | 100% | 48 | 48 | 100% | 6 | 30 | 0 | 0 | 0 |
| Erythromycin | Streptococcus | 90 | 90 | 100% | 29 | 29 | 100% | 90 | 90 | 100% | 54 | 36 | 0 | 0 | 0 |
| Agent | Organism group | Total tested | # EA | % EA | Total Evaluable | # EA of Evaluable | % EA of Evaluable | Total cat. | # CA | % CA | # S | # R | # vmj | # maj | # mi |
| Gentamicin | Enterobacterales | 186 | 186 | 100% | 40 | 40 | 100% | 186 | 185 | 99.5% | 108 | 66 | 0 | 0 | 1 |
| Gentamicin | Non-fermenters | 81 | 81 | 100% | 27 | 27 | 100% | 81 | 81 | 100% | 54 | 21 | 0 | 0 | 0 |
| Imipenem | Staphylococcus | 48 | 48 | 100% | 6 | 6 | 100% | 48 | 48 | 100% | 27 | 21 | 0 | 0 | 0 |
| Imipenem | Non-fermenters | 105 | 104 | 99.0% | 54 | 54 | 100% | 105 | 105 | 100% | 72 | 33 | 0 | 0 | 0 |
| Levofloxacin | Enterobacterales | 186 | 186 | 100% | 47 | 47 | 100% | 186 | 183 | 98.4% | 69 | 99 | 0 | 0 | 3 |
| Levofloxacin | Non-fermenters | 81 | 81 | 100% | 63 | 63 | 100% | 81 | 79 | 97.5% | 42 | 27 | 0 | 0 | 2 |
| Levofloxacin | Staphylococcus | 33 | 33 | 100% | 14 | 14 | 100% | 33 | 32 | 97.0% | 27 | 6 | 0 | 0 | 1 |
| Levofloxacin | Enterococcus | 48 | 48 | 100% | 41 | 41 | 100% | 48 | 47 | 97.9% | 27 | 6 | 0 | 0 | 1 |
| Linezolid | Streptococcus | 90 | 90 | 100% | 90 | 90 | 100% | 90 | 90 | 100% | 87 | 0 | 0 | 0 | 0 |
| Linezolid | Staphylococcus | 48 | 48 | 100% | 48 | 48 | 100% | 48 | 48 | 100% | 48 | 0 | 0 | 0 | 0 |
| Linezolid | Enterococcus | 48 | 48 | 100% | 48 | 48 | 100% | 48 | 47 | 97.9% | 45 | 0 | 0 | 0 | 1 |
| Linezolid | Streptococcus | 90 | 90 | 100% | 0 | 0 | N/A | 90 | 90 | 100% | 90 | 0 | 0 | 0 | 0 |
| Meropenem | Enterobacterales | 186 | 186 | 100% | 26 | 26 | 100% | 186 | 184 | 98.9% | 111 | 72 | 0 | 0 | 2 |
| Meropenem | Non-fermenters | 105 | 105 | 100% | 64 | 64 | 100% | 105 | 105 | 100% | 72 | 27 | 0 | 0 | 0 |
| Moxifloxacin | Staphylococcus | 48 | 48 | 100% | 12 | 12 | 100% | 48 | 48 | 100% | 39 | 6 | 0 | 0 | 0 |
| Nitrofurantoin | Enterobacterales | 186 | 186 | 100% | 134 | 134 | 100% | 186 | 184 | 98.9% | 51 | 90 | 0 | 0 | 2 |
| Nitrofurantoin | Staphylococcus | 48 | 48 | 100% | 2 | 2 | 100% | 48 | 48 | 100% | 48 | 0 | 0 | 0 | 0 |
| Oxacillin | Enterococcus | 48 | 48 | 100% | 27 | 27 | 100% | 48 | 47 | 97.9% | 24 | 15 | 0 | 0 | 1 |
| Oxacillin | Staphylococcus | 48 | 48 | 100% | 13 | 13 | 100% | 48 | 48 | 100% | 21 | 27 | 0 | 0 | 0 |
| Penicillin (Benzyl-penicillin) | Enterococcus | 48 | 48 | 100% | 38 | 38 | 100% | 48 | 48 | 100% | 33 | 15 | 0 | 0 | 0 |
| Penicillin (Benzyl-penicillin) | Streptococcus | 90 | 90 | 100% | 25 | 25 | 100% | 90 | 90 | 100% | 90 | 0 | 0 | 0 | 0 |
| Piperacillin / Tazobactam | Staphylococcus | 48 | 48 | 100% | 11 | 11 | 100% | 48 | 48 | 100% | 12 | 36 | 0 | 0 | 0 |
| Piperacillin / Tazobactam | Enterobacterales | 177 | 177 | 100% | 29 | 29 | 100% | 177 | 176 | 99.4% | 60 | 111 | 0 | 0 | 1 |
| Piperacillin / Tazobactam | Non-fermenters | 105 | 105 | 100% | 52 | 52 | 100% | 105 | 103 | 98.1% | 57 | 30 | 0 | 0 | 2 |
| Quinupristin / Dalfopristin | Staphylococcus | 48 | 48 | 100% | 16 | 16 | 100% | 48 | 48 | 100% | 45 | 3 | 0 | 0 | 0 |
| Agent | Organism group | Total tested | # EA | % EA | Total Evaluable | # EA of Evaluable | % EA of Evaluable | Total cat. | # CA | % CA | # S | # R | # vmj | # maj | # min |
| Tetracycline | Enterobacterales | 186 | 186 | 100% | 54 | 54 | 100% | 186 | 184 | 98.9% | 90 | 87 | 0 | 0 | 2 |
| | Staphylococcus | 48 | 48 | 100% | 7 | 7 | 100% | 48 | 48 | 100% | 30 | 18 | 0 | 0 | 0 |
| | Enterococcus | 48 | 48 | 100% | 0 | 0 | N/A | 48 | 48 | 100% | 15 | 33 | 0 | 0 | 0 |
| | Streptococcus | 42 | 42 | 100% | 0 | 0 | N/A | 42 | 42 | 100% | 39 | 3 | 0 | 0 | 0 |
| Tigecycline | Enterobacterales | 186 | 186 | 100% | 104 | 104 | 100% | 186 | 183 | 98.4% | 144 | 18 | 0 | 0 | 3 |
| | Staphylococcus | 33 | 33 | 100% | 0 | 0 | N/A | 33 | 33 | 100% | 33 | 0 | 0 | 0 | 0 |
| | Enterococcus | 21 | 21 | 100% | 0 | 0 | N/A | 21 | 21 | 100% | 21 | 0 | 0 | 0 | 0 |
| | Streptococcus | 90 | 90 | 100% | 2 | 2 | 100% | 90 | 90 | 100% | 90 | 0 | 0 | 0 | 0 |
| Tobramycin | Enterobacterales | 186 | 186 | 100% | 42 | 42 | 100% | 186 | 185 | 99.5% | 51 | 108 | 0 | 0 | 1 |
| | Non-fermenters | 81 | 81 | 100% | 1 | 1 | 100% | 81 | 80 | 98.8% | 57 | 24 | 0 | 0 | 1 |
| Trimethoprim / Sulfamethoxazole | Enterobacterales | 186 | 186 | 100% | 3 | 3 | 100% | 186 | 186 | 100% | 63 | 123 | 0 | 0 | 0 |
| Vancomycin | Staphylococcus | 48 | 48 | 100% | 35 | 35 | 100% | 48 | 48 | 100% | 45 | 0 | 0 | 0 | 0 |
| | Enterococcus | 48 | 48 | 100% | 18 | 18 | 100% | 48 | 48 | 100% | 30 | 18 | 0 | 0 | 0 |
| | Streptococcus | 90 | 89 | 98.9% | 65 | 65 | 100% | 90 | 90 | 100% | 90 | 0 | 0 | 0 | 0 |

### AST Challenge Study summary of results, antimicrobial agent

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| Group | Incub. time | Total<br>tested | # EA | % EA | Total<br>Evaluable | # EA of<br>Evaluable | % EA of<br>Evaluable | Total<br>cat. | # CA | % CA | # S | # R | # vmj | # maj | # min |
|------------------|-------------|-----------------|------|-------|--------------------|----------------------|----------------------|---------------|------|-------|-----|-----|-------|-------|-------|
| Enterobacterales | 14 h | 1383 | 1382 | 99.9% | 361 | 361 | 100% | 1383 | 1373 | 99.3% | 561 | 735 | 0 | 0 | 10 |
| | 24 h | 1689 | 1688 | 99.9% | 458 | 457 | 99.8% | 1689 | 1672 | 99.0% | 705 | 834 | 0 | 0 | 17 |
| Non-fermenters | 14 h | 420 | 420 | 100% | 200 | 200 | 100% | 420 | 416 | 99.0% | 252 | 126 | 0 | 0 | 4 |
| | 24 h | 510 | 509 | 99.8% | 251 | 251 | 100% | 510 | 504 | 98.8% | 339 | 135 | 0 | 1 | 5 |
| Staphylococcus | 18 h | 594 | 594 | 100% | 179 | 179 | 100% | 594 | 592 | 99.7% | 429 | 153 | 0 | 0 | 2 |
| Enterococcus | 18 h | 453 | 453 | 100% | 221 | 221 | 100% | 453 | 449 | 99.1% | 264 | 141 | 0 | 0 | 4 |
| Streptococcus | 18 h | 942 | 941 | 99.9% | 213 | 213 | 100% | 942 | 941 | 99.9% | 864 | 72 | 0 | 0 | 1 |

#### AST Challenge Study summary of results, organism group

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MICs obtained using Colibrí System for microbial suspensions preparation showed very high agreement with the manual preparation for all the microorganisms group; overall, 1882/1883 evaluable MIC results were within one doubling dilution of the comparator method result and 5947/5991SIR categorizations were in agreement. The overall Essential Agreement of the evaluable MIC results was > 99.9% and the Category Agreement was 99.3%.

### Reproducibility Study

The Reproducibility Study was performed to demonstrate consistency of AST results given by bioMérieux VITEK 2 on microbial suspensions prepared by different Colibrí Systems in different test days.

Culture media showing isolated colonies of different Gram-Positive and Gram-Negative strains were processed on three Colibrí Systems run by three operators over 3 days. Each microorganism was tested with the appropriate antibiotic panel following the analyzer's instructions for use. Each condition was tested in triplicate for a total number of 81 replicates for each combination strainantimicrobial agent.

The MIC results were considered reproducible if they fell within one doubling dilution from the modal value of each combination strain-antimicrobial agent

The results were reproducible for each antimicrobial agent between instruments, operators and days.

| Antibiotic | Colibrí System | Best caseª (%) | Worst case (%) |
|-----------------------------|----------------|-----------------|-----------------|
| Ampicillin-<br>Sulbactam | Instrument 1 | 27/27 (100%) | 27/27 (100%) |
| | Instrument 2 | 27/27 (100%) | 27/27 (100%) |
| | Instrument 3 | 27/27 (100%) | 27/27 (100%) |
| | Combined | 81/81 (100%) | 81/81 (100%) |
| Piperacillin/<br>Tazobactam | Instrument 1 | 134/135 (99.3%) | 133/135 (98.5%) |
|…