Curetis Unyvero LRT Application

DEN170047 · Curetis GmbH · QBH · Apr 3, 2018 · Microbiology

Device Facts

• Record ID: DEN170047
• Device Name: Curetis Unyvero LRT Application
• Applicant: Curetis GmbH
• Product Code: QBH · Microbiology
• Decision Date: Apr 3, 2018
• Decision: DENG
• Submission Type: Direct
• Regulation: 21 CFR 866.3985
• Device Class: Class 2

Indications for Use

The Unyvero LRT Application is a qualitative nucleic acid multiplex test intended for the simultaneous detection and identification of nucleic acid sequences from the following microorganisms and antibiotic resistance markers in endotracheal aspirates from adult hospitalized patients with suspected lower respiratory tract infections. The Unyvero LRT Application performed on the Unyvero System is indicated as an aid in the diagnosis of lower respiratory tract infection in adult hospitalized patients with signs and symptoms of lower respiratory infection: results should be used in conjunction with other clinical and laboratory findings. As tracheal aspirates commonly contain colonizing microorganisms, detection of Unyvero LRT microbial targets does not indicate that the microorganism is the cause of the disease. Unyvero positive results do not rule out co-infection with microorganisms not detected by the Unyvero LRT Application. Negative results do not preclude lower respiratory infection, as the causative agent may be a microorganism not detected by this test. A negative result for any antibiotic resistance marker does not indicate that detected microorganisms are susceptible to applicable antimicrobial agents. Detected resistance markers cannot be definitively linked to specific microorganisms, and may be present in organisms that are not detected by the Unyvero LRT Application such as organisms present as colonizing or normal flora. Microbiology cultures of aspirates should be performed to obtain isolates for species identification and antimicrobial susceptibility testing, to differentiate quantities of identified microorganisms as well as normal flora present in the specimen and to identify potential microorganisms not targeted by the Unyvero LRT Application.

Device Story

Unyvero LRT Application is an in vitro diagnostic, qualitative nucleic acid multiplex test. Input: endotracheal aspirates from adult hospitalized patients. Operation: system processes samples to detect specific microbial nucleic acid sequences and antibiotic resistance markers. Output: identification of presence/absence of targeted microorganisms and resistance genes. Used in hospital settings; operated by trained laboratory personnel. Results assist clinicians in diagnosing lower respiratory tract infections; however, results do not confirm infection causality or antibiotic susceptibility. Clinical decision-making must incorporate culture results and patient history. Benefits include rapid identification of potential pathogens and resistance markers to support clinical management.

Clinical Evidence

Prospective multi-center study (N=603 evaluable specimens) compared Unyvero LRT to standard-of-care culture and composite molecular comparator (PCR/sequencing). Performance for typical microorganisms generally showed >90% sensitivity and >95% specificity vs culture. Atypical analytes validated via contrived specimens. Resistance marker performance evaluated vs molecular comparator; results masked if host microorganism not detected. Study included linkage analysis of resistance markers to culture isolates.

Technological Characteristics

Qualitative nucleic acid multiplex test. Methodology involves specimen processing and molecular detection of specific microbial and resistance marker sequences. System includes instrument, assay-specific reagents, and software for data analysis. Requires adherence to special controls regarding analytical performance (LOD, inclusivity, cross-reactivity, etc.) and design verification.

Indications for Use

Indicated for adult hospitalized patients with suspected lower respiratory tract infections (LRTIs) exhibiting signs and symptoms of lower respiratory infection. For use with endotracheal aspirates. Prescription use only.

Regulatory Classification

Identification

A device to detect and identify microorganisms and associated resistance marker nucleic acids directly from respiratory specimens is an in vitro diagnostic device intended for the detection and identification of microorganisms and associated resistance markers in respiratory specimens collected from patients with signs or symptoms of respiratory infection. The device is intended to aid in the diagnosis of respiratory infection in conjunction with clinical signs and symptoms and other laboratory findings. These devices do not provide confirmation of antibiotic susceptibility since mechanisms of resistance may exist other than those detected by the device.

Special Controls

*Classification.* Class II (special controls). The special controls for this device are:(1) The intended use for the 21 CFR 809.10 labeling 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. (2) The 21 CFR 809.10(b) labeling 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, including all pre-analytical methods for processing of specimens. (ii) Performance characteristics from analytical studies, including, but not limited to, limit of detection, inclusivity, reproducibility, cross reactivity, interfering substances, competitive inhibition, carryover/cross contamination, specimen stability, and linearity, as applicable. (iii) A limiting statement that the device is intended to be used in conjunction with clinical history, signs and symptoms, and results of other diagnostic tests, including culture and antimicrobial susceptibility testing. (iv) A detailed explanation of the interpretation of test results for clinical specimens and acceptance criteria for any quality control testing. (v) A limiting statement that negative results for microorganisms do not preclude the possibility of infection, and should not be used as the sole basis for diagnosis, treatment, or other patient management decisions. (vi) If applicable, a limiting statement that detected microorganisms may not be the cause of lower respiratory tract infection and may be indicative of colonizing or normal respiratory flora. (vii) If applicable, a limiting statement that detection of resistance markers cannot be definitively linked to specific microorganisms and that the source of a detected resistance marker may be an organism not detected by the assay, including colonizing flora. (viii) If applicable, a limiting statement that detection of antibiotic resistance markers may not correlate with phenotypic gene expression. (3) The 21 CFR 809.10(b) labeling and any test report generated by the device must include a limiting statement that negative results for resistance markers do not indicate susceptibility of detected microorganisms. (4) Design verification and validation must include: (i) Performance characteristics from clinical studies that include prospective (sequential) samples and, if appropriate, additional characterized samples. The study must be performed on a study population consistent with 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. Results 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) A detailed device description including the following: (A) Thorough description of the assay methodology including, but not limited to, primer/probe sequences, primer/probe design, and rationale for target sequence selection, as applicable. (B) Algorithm used to generate a final result from raw data (e.g., how raw signals are converted into a reported result). (iii) A detailed description of device software, including, but not limited to, validation activities and outcomes. (iv) As part of the risk management activities, an appropriate end user device training program must be offered as an effort to mitigate the risk of failure from user error.

Submission Summary (Full Text)

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# EVALUATION OF AUTOMATIC CLASS III DESIGNATION FOR Unyvero LRT Application and Unyvero System DECISION SUMMARY

## A. DEN Number:

## DEN170047

- B. Purpose for Submission: De Novo request for evaluation of automatic class III designation for the Unyvero Lower Respiratory Tract (LRT) Application and the Unyvero System.
- C. Measurand: DNA sequences of the following organisms and antibiotic resistance markers:

| Microorganism Targets | Antibiotic Resistance Marker Targets |
|------------------------------|--------------------------------------------|
| Acinetobacter spp. | ctx-M ( <i>bla</i> CTX-M, subgroup 1 only) |
| Chlamydia pneumoniae | kpc ( <i>bla</i> KPC) |
| Citrobacter freundii | mecA |
| Escherichia coli | ndm ( <i>bla</i> NDM) |
| Enterobacter cloacae complex | oxa-23 ( <i>bla</i> OXA-23) |
| Haemophilus influenzae | oxa-24 ( <i>bla</i> OXA-24) |
| Klebsiella oxytoca | oxa-48 ( <i>bla</i> OXA-48) |
| Klebsiella pneumoniae | oxa-58 ( <i>bla</i> OXA-58) |
| Klebsiella variicola | tem ( <i>bla</i> TEM) |
| Legionella pneumophila | vim ( <i>bla</i> VIM) |
| Moraxella catarrhalis | |
| Morganella morganii | |
| Mycoplasma pneumoniae | |
| Proteus spp. | |
| Pseudomonas aeruginosa | |
| Serratia marcescens | |
| Staphylococcus aureus | |
| Stenotrophomonas maltophilia | |
| Streptococcus pneumoniae | |

## D. Type of Test:

Qualitative nucleic acid amplification assay

## E. Applicant:

Curetis GmbH

### F. Proprietary and Established Names:

Unyvero Lower Respiratory Tract (LRT) Application Unyvero System

# G. Regulatory Information:

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# 1. Regulation section:

21 CFR 866.3985

- 2. Classification:
Class II (Special Controls)

- 3. Product code:
QBH

- 4. Panel:
83-Microbiology

# H. Indications for use:

- 1. Indications for use:
The Unyvero LRT Application is a qualitative nucleic acid multiplex test intended for the simultaneous detection and identification of nucleic acid sequences from the following microorganisms and antibiotic resistance markers in endotracheal aspirates from adult hospitalized patients with suspected lower respiratory tract infections.

| Microorganism | Associated antibiotic resistance marker |
|--------------------------------|-----------------------------------------------|
| Acinetobacter spp. a | ctx-Mb, kpc, ndm, oxa-23, oxa-24, oxa-58, vim |
| Chlamydia pneumoniae | - |
| Citrobacter freundii | ctx-Mb, kpc, ndm, oxa-48, vim |
| Enterobacter cloacae complex c | ctx-Mb, kpc, ndm, oxa-48, vim |
| Escherichia coli | ctx-Mb, kpc, ndm, oxa-48, vim |
| Haemophilus influenzae | tem |
| Klebsiella oxytoca | ctx-Mb, kpc, ndm, oxa-48, vim |
| Klebsiella pneumoniae d | ctx-Mb, kpc, ndm, oxa-48, vim |
| Klebsiella variicola | ctx-Mb, kpc, ndm, oxa-48, vim |
| Legionella pneumophila | - |
| Moraxella catarrhalis | - |
| Morganella morganii | ctx-Mb, kpc, ndm, oxa-48, vim |
| Mycoplasma pneumoniae | - |
| Proteus spp. e | ctx-Mb, kpc, ndm, oxa-48, vim |
| Pseudomonas aeruginosa | ctx-Mb, kpc, ndm, vim |
| Serratia marcescens | ctx-Mb, kpc, ndm, oxa-48, vim |
| Staphylococcus aureus | mecA |
| Stenotrophomonas maltophilia | - |
| Streptococcus pneumoniae | - |

ª Acinetobacter spp. includes: A. baumannii, A. calcoaceticus, A. haemolyticus, A. junii, A.

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lwoffii, A. nosocomialis, A. parvus A. pittii, (detected by LRT Application) and A. ursingii (not detected by LRT Application). b ctx-M1 subgroup. & Enterobacter cloacae complex includes: E. asburiae, E. cloacae, E. hormaechei, E. kobei, E. ludwigii, and E. xiangfangensis. d Klebsiella pneumoniae includes two variants: K. pneumoniae (variant 1), and K. quasipneumoniae (variant 2) e Proteus spp. includes P. hauseri, P. mirabilis, P. penneri and P. vulgaris

The Unyvero LRT Application performed on the Unyvero System is indicated as an aid in the diagnosis of lower respiratory tract infection in adult hospitalized patients with signs and symptoms of lower respiratory infection: results should be used in conjunction with other clinical and laboratory findings. As tracheal aspirates commonly contain colonizing microorganisms, detection of Unyvero LRT microbial targets does not indicate that the microorganism is the cause of the disease. Unyvero positive results do not rule out co-infection with microorganisms not detected by the Unyvero LRT Application. Negative results do not preclude lower respiratory infection, as the causative agent may be a microorganism not detected by this test.

A negative result for any antibiotic resistance marker does not indicate that detected microorganisms are susceptible to applicable antimicrobial agents. Detected resistance markers cannot be definitively linked to specific microorganisms, and may be present in organisms that are not detected by the Unyvero LRT Application such as organisms present as colonizing or normal flora.

Microbiology cultures of aspirates should be performed to obtain isolates for species identification and antimicrobial susceptibility testing, to differentiate quantities of identified microorganisms as well as normal flora present in the specimen and to identify potential microorganisms not targeted by the Unyvero LRT Application.

- 2. Special conditions for use statement(s):
- . For in vitro diagnostic use only
- . For prescription use only

Limitations:

- . Antimicrobial resistance can occur via mechanisms other than the resistance markers detected by the Unyvero LRT Assay. Negative results for the LRT resistance markers do not indicate antimicrobial susceptibility of detected organisms.
- . Detected resistance markers cannot be definitively linked to specific microorganism(s), and may be present in organisms that are not detected by the Unyvero LRT assay, including colonizing flora. Standard culture of tracheal aspirates and subsequent testing of the isolated microorganism(s) is necessary to definitively link antimicrobial resistance with a specific microorganism.
- Detection of antibiotic resistance markers in a specimen may not correlate with phenotypic gene expression. Resistance marker results should be used in

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conjunction with final culture and antimicrobial susceptibility testing results.

- . Detected microorganisms may be indicative of colonizing or normal respiratory flora and may not be the causative agent of pneumonia.
(b) (4)

- The LRT Application is a qualitative test and does not provide a quantitative ● value for the microorganisms or antibiotic resistance markers detected in the specimen. Compared to standard of care culture methods, the Unyvero LRT Application is not able to assess the presence or amount of normal flora in the specimen or whether the detected microorganism/antibiotic resistance marker is present in predominating amounts as compared to other microorganisms in the specimen. The LRT Application also does not assess if detected microorganisms are present at clinically relevant concentrations, or are present as colonizing flora.
- . When three or more microorganisms are detected for a specimen, the user is advised to wait for microbiology culture results to verify the predominant microorganism(s) and to assess for the presence of normal respiratory flora.
- Bacterial nucleic acids may be present in the specimen independent of organism ● viability. Detection of organism nucleic acid target(s) does not imply that the corresponding organisms are the causative agents for clinical symptoms.
- . A negative result for the 'atypical' microorganisms (C. pneumoniae, L. pneumophila, and M. pneumoniae) does not exclude the presence of this microorganism in the patient specimen. A positive result should be evaluated in the overall context of the patient's clinical condition and other laboratory results being part of the standard-of-care routine.
- . Due to the small number of positive prospective and archived specimens for certain microorganisms and antibiotic resistance markers, performance characteristics for C. pneumoniae, C. freundii, K. oxytoca, K. variicola, L. pneumophila, M. catarrhalis, M. morganii, M. pneumoniae, and antibiotic resistance markers ctx-M, kpc, ndm, oxa-23, oxa-24, oxa-48, oxa-58, and vim were established primarily using contrived clinical specimens.
- . Based on in-silico analysis and inclusivity wet testing, certain clinically relevant species for some LRT panel microorganism analytes are not detected or detected with reduced sensitivity: Acinetobacter ursingii detection is predicted at reduced sensitivity for the Acinetobacter spp. assay, Enterobacter asburiae detection is predicted at reduced sensitivity for the Enterobacter cloacae complex assay.
- Subtypes included in ctx-M subgroups ctx-M2, ctx-M8, ctx-M9 (ctx-M14), ctx-. M25 and ctx-M45 were not evaluated and are not predicted to be detected by the LRT assay based on in-silico analysis.
- . The mecC variant is not detected by Unyvero LRT and for mecC positive Staphylococcus aureus strains, the assay will generate negative results for mecA.
- . Based on in-silico analysis and exclusivity wet testing. the following LRT panel microorganism assays are expected to cross-react with closely related clinically relevant species: Citrobacter freundii (cross-reactive to C. braakii, Kluyvera georgiana), Escherichia coli (cross-reactive to E. albertii, E. fergusonii and

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Shigella spp. (S. dysenteriae, S. sonnei, S. boydii)), Haemophilus influenzae (cross-reactive to H. haemolyticus), Klebsiella oxytoca (cross-reactive to K. michiganensis).

# 4. Special instrument requirements:

The Unyvero LRT Application is performed on the Unyvero System which includes the Unyvero Lysator, Unyvero Analyzer and Unyvero Cockpit.

# I. Device Description:

The Unyvero LRT Application is a qualitative test that includes specimen processing, genomic bacterial DNA isolation and purification, multiplex PCR and array hybridization and detection. The Unyvero LRT Application performed using the Unyvero System detects specific nucleic acid sequences from microorganisms and resistance markers in tracheal aspirates collected from patients with signs and symptoms of lower respiratory infection.

The Unyvero LRT Application consists of the following components:

- Unyvero LRT Cartridge: Contains DNA isolation and purification reagents, a DNA . isolation column, eight separate PCR chambers with eight corresponding detection arrays. The Cartridge also contains fluorescently-labeled primers, hybridization and wash buffers and oligonucleotide probes for detection of targeted PCR products using array hybridization technology.
- Unyvero T1 Sample Tube: Contains glass beads and buffers to lyse bacteria and liquefy the sample.
- . Unyvero T1 Sample Tube Cap (with Internal Control): Contains proteinase K and a synthetic internal control gene for process monitoring. The T1 Sample Tube Cap seals the Unyvero Sample Tube after which the internal control is combined with each patient specimen. The internal control DNA sequence does not have significant homology to targeted sequences and is amplified independently in each of the eight PCR chambers and the amplified internal control product is hybridized on each array.
- . Unyvero M1 Master Mix: Contains reagents for DNA amplification.
- . Unyvero T1 Transfer Tool: The Transfer tool can be used to transfer viscous specimens from the primary sample container to the Unyvero Sample Tube.

The Unyvero System consists of the following components:

- . Unyvero Lysator: The Lysator lyses the specimen and can process up to four specimens simultaneously in four separate slots.

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- Unyvero Analyzer: The Analyzer automates DNA purification, amplification and detection. Each Analyzer can simultaneously process up to two Unyvero Cartridges with each slot available using random access.
- Unyvero Cockpit: The Cockpit provides the main user interface for the Unyvero System, guides the user through the steps to run the Unvvero LRT Application and automatically generates and displays test results. The Cockpit is equipped with a high-resolution touch screen and a barcode reader.
- Unyvero Sample Tube Holder: The Sample Tube holder holds the Sample Tube securely while the specimen is transferred into the Sample Tube.

Other materials required but not provided:

- Pipette capable of dispensing 180uL
- DNase/RNase free, aerosol resistant pipette tips
- 1mL Luer-Lock svringe ●

# J. Standard/Guidance Document Referenced:

- CLSI EP07-A2. Interference Testing in Clinical Chemistry: Approved Guideline, 2nd ● edition.
- CLSI EP17-A2. Evaluation of Detection Capability for Clinical Laboratory ● Measurement Procedures; Approved Guideline - 2nd edition.
- CLSI M100, Performance Standards for Antimicrobial Susceptibility Testing, 26th ● Edition.

# K. Test Principle:

The Unyvero LRT Application is a qualitative PCR-based assay that detects DNA sequences of microorganisms and associated resistance markers in tracheal aspirate specimens. The assay includes specimen processing (lysis), DNA extraction and isolation, multiplex PCR with eight parallel multiplex endpoint PCR reactions, and qualitative detection of amplified DNA products using hybridization arrays.

A tracheal aspirate specimen is first pipetted into the Unyvero Sample Tube and closed with the Unyvero Sample Tube Cap. Closing the sample tube automatically adds the lysis reagent and the internal control gene template to the specimen. The sample tube which fits into the Unyvero Lysator only if closed with the Unyvero sample tube cap is then placed on the Lysator. After the specimen is lysed in the Lysator, the Sample Tube and Master Mix are loaded into the Unyvero LRT Cartridge which is then inserted into the position assigned by the Unyvero Analyzer for automated processing and analysis.

In the Unyvero LRT Cartridge, the remainder of the testing steps are automated by the Unyvero Analyzer. The lysed specimen is further processed and then transferred onto a

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(b) DNA purification column for nucleic acid (b) (4) DNA is transferred to a chamber, where mixing with the Master Mix takes place. This mixture is distributed into eight separate PCR reaction chambers each containing multiple primer pairs. After amplification, PCR products are hybridized to the corresponding array probes. Each array has been manufactured (b) (4) (b) (4)

(b) (4)

Result data are transferred

to the Unyvero Cockpit for visualization and result printout. A test run is completed after approximately 4.5 hours, and results for panel microorganisms and associated antibiotic resistance markers are displayed on the Unyvero Cockpit screen.

# L. Performance Characteristics (if/when applicable):

# 1. Analytical performance:

# a. Reproducibility:

The reproducibility of the Unyvero LRT Application was assessed using a representative panel of samples prepared in artificial respiratory matrix (ARM, adapted from Dinesh S. D.1) which was used as a surrogate for tracheal aspirate matrix. In a separate analytical study, the Unyvero LRT Application was shown to demonstrate equivalent performance for samples prepared in ARM matrix and samples prepared in natural tracheal aspirate matrix (See Section L.2.b below).

Samples for the reproducibility study were prepared with whole-organism preparations for the following LRT panel analytes: E. coli/tem. P. geruginosa, K. pneumoniae, M. morganii, S. maltophilia and S. aureus/mecA. Sample #1 (REPRO 1) was inoculated with each organism at moderate positive concentrations (~5 x LoD) and Sample #2 (REPRO 2) was inoculated with each organism at low positive concentrations (~1.7 x LoD). For the mec A target, the REPRO 1 and REPRO2 panel members had concentrations of 12.5 x LoD and 4.2 x LoD respectively. For the tem target, the REPRO1 and REPRO2 panel members had concentrations of 6.2 and 2.1 x LoD respectively. Test runs were independently performed by three different operators. Each operator was assigned one of three Unyvero systems, each consisting of 1 Unyvero Cockpit, 2 Unyvero Lysators and at least 4 Unyvero Analyzers. A total of 90 replicate samples were tested for each panel member with each operator testing samples in triplicate on a minimum of five testing days. Positive and negative controls were also tested daily by each operator. The organisms and concentrations used to prepare study samples are presented in Table 1 below. Study results are presented in Table 2.

Table 1: Microorganisms for Reproducibility Study ATCC Strain | LoD (CFU/mL) | Repro1/x-fold LoD | Repro2/x-fold LoD Analytes

¹ Dinesh, Artificial Sputum Medium. Protocol Exchange (2010)

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| E. coli | 35218 | 7.50E+04 | 3.75E+05 | 5.0 | 1.28E+05 | 1.7 |
|------------------|--------|----------|----------|------|----------|-----|
| K. pneumoniae V2 | 700603 | 5.00E+04 | 2.50E+05 | 5.0 | 8.50E+04 | 1.7 |
| M. morganii | 25830 | 5.00E+05 | 2.50E+06 | 5.0 | 8.50E+05 | 1.7 |
| P. aeruginosa | 10145 | 5.00E+04 | 2.50E+05 | 5.0 | 8.50E+04 | 1.7 |
| S. aureus | 33591 | 5.00E+06 | 2.50E+07 | 5.0 | 8.50E+06 | 1.7 |
| S. maltophilia | 13637 | 3.00E+04 | 1.50E+05 | 5.0 | 5.10E+04 | 1.7 |
| mec A | 33591 | 2.00E+06 | 2.50E+07 | 12.5 | 8.50E+06 | 4.3 |
| tem | 35218 | 6.00E+04 | 3.75E+05 | 6.3 | 1.28E+05 | 2.1 |

Table 2: Reproducibility study results, stratified by target, concentration and operator

| | | REPRO1 (5x LoD) | | | | REPRO 2 (1.7x LoD) | | | |
|-------------------------------------------|--------|-----------------|-------|-------|--------------|--------------------|-------|-------|--------------|
| analyte | user | x-fold<br>LoD | Pos. | [%] | 95 % CI | x-<br>fold<br>LoD | Pos. | [%] | 95 % CI |
| E. coli<br>ATCC 35218 | user 1 | 5 | 30/30 | 100.0 | 88.7 - 100.0 | 1.7 | 29/29 | 100.0 | 88.3 - 100.0 |
| | user 2 | | 30/30 | 100.0 | 88.7 - 100.0 | | 29/30 | 96.7 | 83.3 - 99.4 |
| | user 3 | | 29/30 | 96.7 | 83.3 - 99.4 | | 29/30 | 96.7 | 83.3 - 99.4 |
| | all | | 89/90 | 98.9 | 94.0 - 99.8 | | 87/89 | 97.8 | 92.2 - 99.4 |
| K. pneumoniae<br>variant 2<br>ATCC 700603 | user 1 | 5 | 21/29 | 72.4 | 54.3 - 85.3 | 1.7 | 14/30 | 46.7 | 30.2 - 63.9 |
| | user 2 | | 25/29 | 86.2 | 69.4 - 94.5 | | 15/30 | 50.0 | 33.2 - 66.8 |
| | user 3 | | 24/30 | 80.0 | 62.7 - 90.5 | | 19/29 | 65.5 | 47.3 - 80.1 |
| | all | | 70/88 | 79.5 | 70.0 - 86.7 | | 48/89 | 53.9 | 43.6 - 63.9 |
| mecA<br>(from: S. aureus<br>ATCC 33591) | user 1 | 12.5 | 30/30 | 100.0 | 88.7 - 100.0 | 4.2 | 25/29 | 86.2 | 69.4 - 94.5 |
| | user 2 | | 27/30 | 90.0 | 74.4 - 96.5 | | 29/30 | 96.7 | 83.3 - 99.4 |
| | user 3 | | 29/30 | 96.7 | 83.3 - 99.4 | | 29/30 | 96.7 | 83.3 - 99.4 |
| | all | | 86/90 | 95.6 | 89.1 - 98.3 | | 83/89 | 93.3 | 86.1 - 96.9 |
| M. morganii<br>ATCC 25830 | user 1 | 5 | 28/29 | 96.6 | 82.8 - 99.4 | 1.7a | 21/30 | 70.0 | 52.1 - 83.3 |
| | user 2 | | 27/29 | 93.1 | 78.0 - 98.1 | | 23/30 | 76.7 | 59.1 - 88.2 |
| | user 3 | | 28/30 | 93.3 | 78.7 - 98.2 | | 24/29 | 82.8 | 65.5 - 92.4 |
| | all | | 83/88 | 94.3 | 87.4 - 97.5 | | 68/89 | 76.4 | 66.6 - 84.0 |
| P. aeruginosa<br>ATCC 10145 | user 1 | 5 | 30/30 | 100.0 | 88.7 - 100.0 | 1.7 | 30/30 | 100.0 | 88.7 - 100.0 |
| | user 2 | | 30/30 | 100.0 | 88.7 - 100.0 | | 28/30 | 93.3 | 78.7 - 98.2 |
| | user 3 | | 30/30 | 100.0 | 88.7 - 100.0 | | 30/30 | 100.0 | 88.7 - 100.0 |
| | all | | 90/90 | 100.0 | 95.9 - 100.0 | | 88/90 | 97.8 | 92.3 - 99.4 |
| S. aureus<br>ATCC 33591 | user 1 | 5 | 28/30 | 93.3 | 78.7 - 98.2 | 1.7 | 30/30 | 100.0 | 88.7 - 100.0 |
| | user 2 | | 30/30 | 100.0 | 88.7 - 100.0 | | 26/30 | 86.7 | 70.3 - 94.7 |
| | user 3 | | 30/30 | 100.0 | 88.7 - 100.0 | | 29/30 | 96.7 | 83.3 - 99.4 |
| | all | | 88/90 | 97.8 | 92.3 - 99.4 | | 85/90 | 94.4 | 87.6 - 97.6 |
| S. maltophilia<br>ATCC 13637 | user 1 | 5 | 30/30 | 100.0 | 88.7 - 100.0 | 1.7 | 29/30 | 96.7 | 83.3 - 99.4 |
| | user 2 | | 28/30 | 93.3 | 78.7 - 98.2 | | 28/30 | 93.3 | 78.7 - 98.2 |
| | user 3 | | 30/30 | 100.0 | 88.7 - 100.0 | | 29/30 | 96.7 | 83.3 - 99.4 |
| | all | | 88/90 | 97.8 | 92.3 - 99.4 | | 86/90 | 95.6 | 89.1 - 98.3 |
| tem<br>(from: E. coli<br>ATCC 35218) | user 1 | 6.2 | 29/30 | 96.7 | 83.3 - 99.4 | 2.1 | 29/29 | 100.0 | 88.3 - 100.0 |
| | user 2 | | 29/30 | 96.7 | 83.3 - 99.4 | | 22/30 | 73.3 | 55.6 - 85.8 |
| | user 3 | | 30/30 | 100.0 | 88.7 - 100.0 | | 26/30 | 86.7 | 70.3 - 94.7 |
| | all | | 88/90 | 97.8 | 92.3 - 99.4 | | 77/89 | 86.5 | 77.9 - 92.1 |

ª M. morganii was also evaluated at 0.6x LoD in PBS matrix in an initial reproducibility study. Results for this panel member were acceptable based on the microorganism concentration tested (e.g., positivity: 77/88, 87.5% detection for samples prepared at below LoD concentrations).

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Testing of reproducibility samples generated acceptable results for all analytes except for K. pneumoniae and M. morganii for which positivity rates were lower than expected for both the 5x LoD and 1.7x LoD panel members. The reason for these unexpected results was likely due to inaccurate quantitation of organism stocks used for sample preparation. It was noted that testing by different operators/sites generated equivalent results for these analytes (i.e., similarly lower than expected percent positivity at each testing site).

To further evaluate the reproducibility of the Unyvero LRT assay for detection of these two organism targets, ten additional sample replicates for both K. pneumoniae and M. morganii were prepared using freshly grown and quantitated organism suspensions with final sample concentrations of 5x LoD. For both microorganisms, a positivity rate of 90% was observed with 9/10 samples generating positive results.

Out of 14 positive quality control runs with samples containing moderate positive analyte concentrations (~8x LoD). one quality control sample was false negative for K. pneumoniae and one sample was false negative for M. morganii.

Three false positive results were observed during the study; two false positive results in reproducibility test sample runs (S. pneumoniae, vim) and one false positive in a positive control run (M. catarrhalis).

Partially invalid results (one PCR chamber only) were observed in 8/180 (4.4%) reproducibility test runs, 2/14 (1.4%) negative quality control runs, and 0/14 positive quality control runs.

Results from the reproducibility study were further evaluated based on Unyvero signal output levels (RBU) for each target and shown in Table 3 below.

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| analyte | user | x-fold<br>LoD | Avg<br>Signal | % CV | Range | x-fold<br>LoD | Avg<br>Signal | % CV | Range |
|----------------|--------|---------------|---------------|--------|---------|---------------|---------------|---------|---------|
| | user 1 | 5 | (b) (4) | b) (4) | (b) (4) | 1.7 | (b) (4) | (b) (4) | (b) (4) |
| E. coli | user 2 | | | | | | | | |
| ATCC 35218 | user 3 | | | | | | | | |
| | all | | | | | | | | |
| K. pneumoniae | user 1 | 5 | (b) (4) | | (b) (4) | 1.7b | (b) (4) | | (b) (4) |
| variant 2 | user 2 | | | | | | | | |
| ATCC 700603 | user 3 | | | | | | | | |
| | all | | | | | | | | |
| mecA | user 1 | 12.5 | (b) (4) | | (b) (4) | 4.2 | (b) (4) | | (b) (4) |
| (from: | user 2 | | | | | | | | |
| S. aureus | user 3 | | | | | | | | |
| ATCC 33591) | all | | | | | | | | |
| | user 1 | 5 | (b) (4) | | (b) (4) | 1.7 | (b) (4) | | (b) (4) |
| M. morganii | user 2 | | | | | | | | |
| ATCC 25830 | user 3 | | | | | | | | |
| | all | | | | | | | | |
| | user 1 | 5 | (b) (4) | | (b) (4) | 1.7 | (b) (4) | | (b) (4) |
| P. aeruginosa | user 2 | | | | | | | | |
| ATCC 10145 | user 3 | | | | | | | | |
| | all | | | | | | | | |
| | user 1 | 5 | (b) (4) | | (b) (4) | 1.7 | (b) (4) | | (b) (4) |
| S. aureus | user 2 | | | | | | | | |
| ATCC 33591 | user 3 | | | | | | | | |
| | all | | | | | | | | |
| | user 1 | 5 | (b) (4) | | (b) (4) | 1.7 | (b) (4) | | (b) (4) |
| S. maltophilia | user 2 | | | | | | | | |
| ATCC 13637 | user 3 | | | | | | | | |
| | all | | | | | | | | |
| tem | user 1 | 6.2 | (b) (4) | | (b) (4) | 2.1 | (b) (4) | | (b) (4) |
| (from: E. coli | user 2 | | | | | | | | |
| ATCC 35218) | user 3 | | | | | | | | |
| | all | | | | | | | | |

Table 3: Reproducibility study, Quantitative analysis of Unyvero signal levels

- b. Linearity/assay reportable range:
Not applicable.

- c. Traceability, Stability, Expected values (controls, calibrators, or methods):
Internal Control: An internal control consisting of a synthetic DNA sequence without homology to LRT microorganism or resistance marker target sequences is separately processed in each of the eight Unyvero LRT Cartridge PCR chambers. The internal control allows for assessment of DNA purification, amplification, array hybridization, and detection steps. The internal control monitors for the presence of PCR inhibitors

{10}------------------------------------------------

in the specimens and enables the system to detect any failures in the testing process that could potentially result in an incorrect test result. Results generated from Individual PCR chambers are considered valid if successful amplification and detection is completed for either the internal control or any microorganism/resistance marker target in the multiplex PCR reaction.

## External Controls

Positive and negative controls were run daily at each of the testing sites during the clinical study. Testing included (b) (4)

External controls are not provided with the Unyvero LRT Application: however. testing of external positive and negative controls are recommended in the assay labeling. Controls may consist of previously characterized positive samples or negative samples spiked with well characterized microorganisms. Previously characterized negative samples may be used as negative controls. External controls should be used in accordance with local, state, and/or federal regulations, accreditation requirements and individual laboratory's quality control policies, as applicable.

## Specimen Stability

The Unyvero LRT instructions for use indicates that it is acceptable to store tracheal aspirate specimens at 2-8°C for up to 24 hours prior to testing. To evaluate effects on assay performance for 24 hour refrigerated storage prior to starting an LRT test, an analysis was performed for prospectively tested clinical tracheal aspirate specimens that were tested with the LRT application at various times of storage up to 24 hours. Both qualitative performance (compared to culture) and semi-quantitative Unyvero LRT signal levels for positive analytes were assessed between groups stored for different time periods. Results for specimen which were stored for times close to the 24-hour sample storage time limit did not show a significant difference in clinical performance or a significant difference in assay signal as compared to specimens that were tested immediately after collection.

The sample storage recommendations for the LRT application are consistent with those for traditional culture2 which include storage and transport of tracheal aspirate specimens at 2-8°C for up to 24 hours. The analysis of the clinical specimen results together with the same recommendation for tracheal aspirate culture support the claim

² Baron, E. J. et al. A guide to utilization of the microbiology laboratory for diagnosis of infectious diseases: 2013 recommendations by the Infectious Diseases Society of America (IDSA) and the American Society for Microbiology (ASM)(a). Clin. Infect. Dis. Off. Publ. Infect. Dis. Soc. An. 57, (2013).

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of refrigerated storage of tracheal aspirates for up to 24 hours prior to testing with the Unyvero LRT Application.

# d. Detection limit:

A study was conducted to determine the Limit of Detection (LoD) of the Unyvero LRT Application for each targeted analyte. Samples were prepared in Artificial Respiratory Matrix (ARM) which was used as a surrogate for tracheal aspirate matrix. Samples prepared in ARM matrix were shown to generate equivalent results to samples prepared in natural tracheal aspirate matrix (See Section L.2.b below).

For the LoD study, samples were inoculated with multiple organisms (i.e., multispiked). To mitigate potential for competitive inhibition, sample compositions were designed to ensure that only one or two analytes were amplified in each multiplex PCR chamber.

LoD estimates were initially determined by testing small numbers of replicates at multiple different organism concentrations. The LoD for each analyte was then confirmed by testing 20 replicates at the estimated LoD concentration. The final LoD is defined as the lowest concentration (reported as CFU/mL, IFU/mL, or CCU/mL) of sample for which ≥95% of sample replicates generate positive results. Tables 4 and 5 below summarize the confirmed LoDs for each LRT panel microorganism and antibiotic resistance marker.

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| LRT Panel Microorganism | ID of Tested Reference<br>Strain | LoD Concentration<br>[CFU/mL, or other as indicated in<br>footnote] |
|-------------------------------------|----------------------------------------------|---------------------------------------------------------------------|
| Acinetobacter spp. | ATCC 19606 ( <i>A.</i><br><i>baumannii</i> ) | 1x105 |
| Chlamydia pneumoniae (in IFU/mL) a | ATCC VR-2282 | 1.5x104 |
| Citrobacter freundii | ATCC 8090 | 2x105 |
| Escherichia coli | ATCC 11775 | 7.5x104 |
| Enterobacter cloacae complex | ATCC 13047 ( <i>E. cloacae</i> ) | 5x106 |
| Haemophilus influenzae | ATCC 33391 | 2x106 |
| Klebsiella oxytoca | ATCC 13182 | 2x105 |
| Klebsiella pneumoniae Variant 1 | ATCC 13883 | 4.5x105 |
| Klebsiella pneumoniae Variant 2 | ATCC 700603 | 5x104 |
| Klebsiella variicola | ATCC BAA-830 | 1x105 |
| Legionella pneumophila | ATCC 33152 | 2x106 |
| Moraxella catarrhalis | ATCC 25238 | 8x105 |
| Morganella morganii | ATCC 25830 | 5x105 |
| Mycoplasma pneumoniae (in CCU/mL) b | ATCC 29085 | 1x105 |
| Proteus mirabilis | ATCC 29906 | 1x105 |
| Proteus vulgaris | ATCC 29905 | 6x105 |
| Pseudomonas aeruginosa | ATCC 10145 | 5x104 |
| Serratia marcescens | ATCC 13880 | 1x105 |
| Staphylococcus aureus | ATCC 12600 | 5x106 |
| Stenotrophomonas maltophilia | ATCC 13637 | 3x104 |
| Streptococcus pneumoniae | ATCC 49619 | 5x105 |

Table 4: Limit of detection for LRT panel microorganisms

a IFU: inclusion-forming units

b CCU: color-changing units

| LRT Panel Antibiotic Resistance Marker | ID of Tested Reference Strain | LoD Concentration [CFU/mL] |
|----------------------------------------|-------------------------------------|----------------------------|
| ctx-M | NRZ-00751 ( <i>K. pneumoniae</i> ) | 1x105 |
| kpc | NRZ-00281 ( <i>E. coli</i> ) | 5x105 |
| <i>mecA</i> | ATCC 33591 ( <i>S. aureus</i> ) | 2x106 |
| ndm | JMI 50067 ( <i>E. coli</i> ) | 5x104 |
| oxa-23 | NCTC 13301 ( <i>A. baumannii</i> ) | 2x107 |
| oxa-24 | NCTC 13302 ( <i>A. baumannii</i> ) | 5x105 |
| oxa-48 | NCTC 13442 ( <i>K. pneumoniae</i> ) | 2x106 |
| oxa-58 | NCTC 13305 ( <i>A. baumannii</i> ) | 7.5x105 |
| tema | NCTC 13351 ( <i>E. coli</i> ) | 6x104 |
| vim | DSM-24600 ( <i>P. aeruginosa</i> ) | 5x104 |

Table 5: Limit of detection for I RT panel antibiotic resistance markers

a Although the LRT Application reports tem only with H. influenzae, LoD was determined with an E. coli strain positive for tem. Note that inclusivity testing was successfully performed with two tem positive H. influenzae strains.

## e. Analytical Reactivity (Inclusivity)

The inclusivity study of the LRT Application was assessed using samples inoculated at low positive concentrations with well-characterized bacterial isolates. Most strains were evaluated in samples prepared at <3x LoD. Testing was performed in duplicate

{13}------------------------------------------------

and if 100% detection was not observed during initial testing, additional samples prepared with higher organism concentrations were evaluated.

Analyte LoDs were established using ARM as a surrogate for aspirate matrix. Inclusivity testing was performed with contrived samples prepared in PBS. Analytical testing of PBS and ARM matrices demonstrated that the LoDs for the two matrices are comparable for most analytes; however, for a few analytes, PBS and ARM LoDs differed by a factor between 0.3-3. The differences in LoD between the two matrices for these few analytes was not considered significant and did not raise concerns that the stated reactivity would differ when testing clinical specimens.

## Inclusivity/Microorganism Targets:

Inclusivity testing for samples prepared with microorganism concentrations at <3x LoD generated 100% detection for the LRT microorganism targets except for the following microorganisms for which one or more strains were not detected or were detected only at higher than LoD concentrations (also bolded in table below):

- One of four strains of Proteus mirabilis (detected at 5x LoD and not detected at ● lower concentrations). Sequencing of the isolate revealed sequence mismatches in a detection probe.
- . One of three strains of Proteus vulgaris (detected at 5x LoD and not detected at lower concentrations). Sequencing of the isolate revealed sequence mismatches in a detection probe.
- . One of six strains of Moraxella catarrhalis (detected at 3.8x LoD and not detected at lower concentrations)
- . One of seven strains of Serratia marcescens (1/2 replicates detected at 5x LoD and 0/2 replicates detected at 10x LoD)
- Two of six strains of Stenotrophomonas maltophilia (<100% detection at all concentrations tested):
- Strain DSM-50173 was positive in 3/4 replicates prepared at 1.7 x LoD । and 1/2 replicates prepared at 6.8x LoD; sequencing of this strain revealed multiple mismatches for one internal probe.
- -Strain DSM-21874 was positive in 2/4 replicates prepared at 1.7 x LoD and 0/2 replicates at 6.8x LoD; sequencing of this strain did not reveal any mismatches to primers or probes. Testing was repeated for this strain at 1x LoD (0/2), 1.7x (0/2) and at 3.3x LoD for which 2/2 sample replicates generated the expected positive result.

The following limitation is included in the Unyvero LRT labeling:

- . Based on in-silico analyses, some strains may either not be detected or be detected with reduced sensitivity due to variations in targeted sequences for the following microorganisms: C freundii. K. oxytoca. L. pneumophilia. M. morganii. Based on wet testing for Proteus spp., and S. marcescens and based

{14}------------------------------------------------

on in-silico/wet testing for M. catarrhalis and S. maltophilia some strains may be detected with reduced sensitivity due to variations in targeted sequences.

Results from inclusivity testing are shown in Table 6.

| Strain | Strain ID | Test Conc.<br>[CFU/mL] | LoD<br>factora | # Positive/<br># Tests |
|-----------------------------------------------------------------------|-----------------------------|-------------------------------------------|------------------------------------|---------------------------------|
| Acinetobacter baumannii | ATCC 19606 | 8x104 | 0.8x | 2/2 |
| Acinetobacter baumannii | NCTC 13305 | 8x104 | 0.8x | 2/2 |
| Acinetobacter baumannii | NCTC 13301 | 8x104 | 0.8x | 2/2 |
| Acinetobacter baumannii | Micromyx 6334 | 8x104 | 0.8x | 2/2 |
| Acinetobacter baumannii | Micromyx 6153 | 8x104 | 0.8x | 2/2 |
| Acinetobacter baumannii | Micromyx 6149 | 1.5x105 | 1.5x | 4/4 |
| Acinetobacter baumannii | Micromyx 4410 | 8x104 | 0.8x | 2/2 |
| Acinetobacter baumannii | JMI 49755 | 1x105 | 1x | 2/2 |
| Acinetobacter baumannii | NRZ-00449 | 8x104 | 0.8x | 2/2 |
| Acinetobacter baumannii | NRZ-00518 | 8x104 | 0.8x | 2/2 |
| Acinetobacter baumannii | UCLA A4 | 8x104 | 0.8x | 2/2 |
| Acinetobacter baumannii | UCLA A5 | 8x104 | 0.8x | 2/2 |
| Acinetobacter calcoaceticus | ATCC 23055 | 8x104 | 0.8x | 2/2 |
| Acinetobacter lwoffii | ATCC 15309 | 8x104 | 0.8x | 2/2 |
| Acinetobacter haemolyticus | ATCC 17906 | 8x104 | 0.8x | 2/2 |
| Chlamydia pneumoniae | ATCC VR-1310 | 4.5x104 b<br>[IFU/mL] | 3x | 2/2 |
| Chlamydia pneumoniae | ATCC VR-2282 | 4.5x104<br>[IFU/mL] | 3x | 2/2 |
| Chlamydia pneumoniae | ATCC 53592 | 4.5x104<br>[IFU/mL] | 3x | 2/2 |
| Citrobacter freundii | ATCC 8090 | 2x105 | 1x | 6/6 |
| Citrobacter freundii | ATCC 43864 | 3x105 | 1.5x | 2/2 |
| Citrobacter freundii | NCTC 8581 | 2x105 | 1x | 2/2 |
| Citrobacter freundii | NCTC 9750 | 5x105 | 2.5x | 2/2 |
| Citrobacter freundii | NRZ-00452 | 5x105 | 2.5x | 2/2 |
| Citrobacter freundii | UCLA C1 | 2x105 | 1x | 2/2 |
| Citrobacter freundii | UCLA C5 | 6x105 | 3x | 2/2 |
| Enterobacter cloacae | ATCC 13047 | 1.5x107 | 3x | 2/2 |
| Enterobacter cloacae | ATCC 23355 | 3x106 | 0.6x | 2/2 |
| Enterobacter cloacae | ATCC 49141 | 6x105 | 0.1x | 2/2 |
| Enterobacter cloacae | ATCC BAA-2468 | 6x105 | 0.1x | 2/2 |
| Enterobacter cloacae | JMI 46239 | 1.5x107 | 3x | 2/2 |
| Enterobacter cloacae | NRZ-00239 | 5x106 | 1x | 2/2 |
| Enterobacter cloacae ssp. dissolvens | ATCC 23373 | 5x106 | 1x | 2/2 |
| Enterobacter hormaechei | ATCC 49162 | 5x106 | 1x | 2/2 |
| Strain | Strain ID | Test Conc.<br>[CFU/mL] | LoD<br>factora | # Positive/<br># Tests |
| <i>Enterobacter asburiae</i> | ATCC 35953 | 1.5x107 | 3x | 2/2 |
| <i>Escherichia coli</i> | ATCC 11775 | 1x105 | 1.3x | 2/2 |
| <i>Escherichia coli</i> | ATCC 25922 | 1x105 | 1.3x | 2/2 |
| <i>Escherichia coli</i> | ATCC 35218 | 1x105 | 1.3x | 4/4 |
| <i>Escherichia coli</i> | ATCC BAA-2523 | 1x105 | 1.3x | 2/2 |
| <i>Escherichia coli</i> | NCTC 13351 | 5x104 | 0.7x | 2/2 |
| <i>Escherichia coli</i> | NCTC 13476 | 1x105 | 1.3x | 2/2 |
| <i>Escherichia coli</i> | JMI 50067 | 6x104 | 0.8x | 2/2 |
| <i>Escherichia coli</i> | NRZ-00176 | 1x105 | 1.3x | 2/2 |
| <i>Escherichia coli</i> | NRZ-00222 | 6x104 | 0.8x | 2/2 |
| <i>Escherichia coli</i> | NRZ-00281 | 1.8x105 | 2.4x | 2/2 |
| <i>Haemophilus influenzae</i> (serotype a) | ATCC 9006 | 2x106 | 1x | 2/2 |
| <i>Haemophilus influenzae</i> (serotype c) | ATCC 9007 | 6x106 | 3x | 2/2 |
| <i>Haemophilus influenzae</i> (serotype b) | ATCC 10211 | 6x106 | 3x | 2/2 |
| <i>Haemophilus influenzae</i> (serotype b) | ATCC 49247 | 6x106 | 3x | 2/2 |
| <i>Haemophilus influenzae</i><br>(non-typeable/non-capsulated) | ATCC 33391 | 6x106 | 3x | 2/2 |
| <i>Haemophilus influenzae</i> (serotype b) | ATCC 49766 | 6x106 | 3x | 2/2 |
| <i>Haemophilus influenzae</i> (serotype b) | NCTC 8468 | 2x106 | 1x | 2/2 |
| <i>Klebsiella oxytoca</i> | ATCC 13182 | 8x104 | 0.4x | 6/6 |
| <i>Klebsiella oxytoca</i> | ATCC 43863 | 8x104 | 0.4x | 2/2 |
| <i>Klebsiella oxytoca</i> | ATCC 8724 | 1.6x105 | 0.8x | 2/2 |
| <i>Klebsiella oxytoca</i> | ATCC 49131 | 8x104 | 0.4x | 2/2 |
| <i>Klebsiella oxytoca</i> | NCIMB 12819 | 4x105 | 2x | 2/2 |
| <i>Klebsiella oxytoca</i> | NRZ-22060 | 8x104 | 0.4x | 2/2 |
| <i>Klebsiella pneumoniae</i> | ATCC 13883 | 4x105 | 0.9x | 4/4 |
| <i>Klebsiella pneumoniae</i> | NCTC 13439 | 1x106 | 2.2x | 2/2 |
| <i>Klebsiella pneumoniae</i> | NCTC 13440 | 6x105 | 1.3x | 2/2 |
| <i>Klebsiella pneumoniae</i> | NCTC 13442 | 4x105 | 0.9x | 2/2 |
| <i>Klebsiella pneumoniae</i> | NCTC 13443 | 4x105 | 0.9x | 2/2 |
| <i>Klebsiella pneumoniae</i> | Micromyx 4653 | 8x105 | 1.8x | 4/4 |
| <i>Klebsiella pneumoniae</i> | Micromyx 4676 | 1x106 | 2.2x | 2/2 |
| <i>Klebsiella pneumoniae</i> | JMI 49831 | 6x105 | 1.3x | 2/2 |
| <i>Klebsiella pneumoniae</i> | JMI 49767 | 9x105 | 2x | 2/2 |
| <i>Klebsiella pneumoniae</i> | NRZ-00002 | 4x105 | 0.9x | 2/2 |
| <i>Klebsiella pneumoniae</i> | NRZ-00103 | 1.5x106 | 3.3x | 2/2 |
| <i>Klebsiella pneumoniae</i> | NRZ-00223 | 9x105 | 2x | 4/4 |
| <i>Klebsiella pneumoniae</i> | NRZ-00249 | 6x105 | 1.3x | 2/2 |
| <i>Klebsiella pneumoniae</i> | NRZ-00472 | 9x105 | 2x | 2/2 |
| <i>Klebsiella pneumoniae</i> | NRZ-00751 | 9x105 | 2x | 2/2 |
| <i>Klebsiella pneumoniae</i> variant II ( <i>K. quasipneumoniae</i> ) | ATCC 700603 | 6x105 | 1.3x | 2/2 |
| Strain | Strain ID | Test Conc.<br>[CFU/mL] | LoD<br>factora | # Positive/<br># Tests |
| Klebsiella variicola | ATCC BAA-830 | 3x105 | 3x | 2/2 |
| Klebsiella variicola | clinical strain 1 | 3.9x105 | 3.9x | 2/2 |
| Klebsiella variicola | clinical strain 2 | 2.6x105 | 2.6x | 2/2 |
| Klebsiella variicola | clinical strain 3 | 1.5x105 | 1.5x | 2/2 |
| Klebsiella variicola | clinical strain 4 | 2.4x105 | 2.4x | 3/3 |
| Klebsiella variicola | clinical strain 5 | 1.5x105 | 1.5x | 2/2 |
| Legionella pneumophila (serotype 1) | ATCC 33152 | 8x105 | 0.4x | 2/2 |
| Legionella pneumophila (serotype 2) | ATCC 33154 | 6x106 | 3x | 2/2 |
| Legionella pneumophila (serotype 3) | ATCC 33155 | 4x105 | 0.2x | 2/2 |
| Legionella pneumophila (serotype 6) | ATCC 33215 | 2x106 | 1x | 2/2 |
| Legionella pneumophila (serotype 8) | ATCC 35096 | 4x105 | 0.2x | 2/2 |
| Legionella pneumophila (serotype 10) | ATCC 43283 | 4x105 | 0.2x | 2/2 |
| Legionella pneumophila | UCLA L1 | 4x105 | 0.2x | 2/2 |
| Legionella pneumophila | UCLA L5 | 4x105 | 0.2x | 2/2 |
| Legionella pneumophila | UCLA L6 | 4x105 | 0.2x | 2/2 |
| Moraxella catarrhalis | ATCC 25238 | 2x106 | 2.5x | 0/2 |
| Moraxella catarrhalis | ATCC 43617 | 4x105 | 0.5x | 2/2 |
| Moraxella catarrhalis | ATCC 8176 | 4x105 | 0.5x | 2/2 |
| Moraxella catarrhalis | ATCC 25240 | 4x105 | 0.5x | 2/2 |
| Moraxella catarrhalis | ATCC 23246 | 4x105 | 0.5x | 2/2 |
| Moraxella catarrhalis | ATCC 49143 | 2x106 | 2.5x | 2/2 |
| Morganella morganii | ATCC 8019 | 1x105 | 0.2x | 2/2 |
| Morganella morganii | ATCC 25829 | 5x105 | 1x | 2/2 |
| Morganella morganii | ATCC 25830 | 1x105 | 0.2x | 2/2 |
| Morganella morganii spp. sibonii | ATCC 49948 | 1.5x106 | 3x | 2/2 |
| Mycoplasma pneumoniae | ATCC 29085 | 3x105 CCU/mLc | 3x | 2/2 |
| Mycoplasma pneumoniae | ATCC 29343 | 3x105 CCU/mLc | 3x | 2/2 |
| Mycoplasma pneumoniae | ATCC 15492 | 3x106 CFU/mL | 3x | 2/2 |
| Mycoplasma pneumoniae | ATCC 15531 | 3x106copies/mL | 3x | 2/2 |
| Proteus mirabilis | ATCC 12453 | 4x104 | 0.4x | 2/2 |
| Proteus mirabilis | ATCC 14153 | 3x105 | 3x | 2/2 |
| Proteus mirabilis | ATCC 25933 | 4x105 | 4x | 0/2 |
| Proteus mirabilis | ATCC 25933 | 5x105 | 5x d | 2/2 |
| Proteus mirabilis | ATCC 29906 | 3x105 | 3x | 4/4 |
| Proteus vulgaris | ATCC 6380 | 2.4x106 | 4x | 0/2 |
| Proteus vulgaris | ATCC 6380 | 3x106 | 5x d | 2/2 |
| Proteus vulgaris | ATCC 8427 | 4x104 | 0.1x | 2/2 |
| Proteus vulgaris | ATCC 29905 | 1.8x106 | 3x | 4/4 |
| Proteus hauseri | ATCC 700826 | 5x104e | 0.1x | 2/2 |
| Proteus penneri | ATCC 33519 | 5x104e | 0.1x | 2/2 |
| Pseudomonas aeruginosa | ATCC 10145 | 1x105 | 2x | 2/2 |
| Strain | Strain ID | Test Conc.<br>[CFU/mL] | LoD<br>factora | # Positive/<br># Tests |
| Pseudomonas aeruginosa | ATCC 27853 | 1x105 | 2x | 2/2 |
| Pseudomonas aeruginosa | DSM-24600 | 5x104 | 1x | 4/4 |
| Pseudomonas aeruginosa | NCTC 13437 | 2x104 | 0.4x | 4/4 |
| Pseudomonas aeruginosa | Micromyx 2562 | 1x105 | 2x | 4/4 |
| Pseudomonas aeruginosa | NRZ-00196 | 2x104 | 0.4x | 2/2 |
| Pseudomonas aeruginosa | NRZ-03961 | 5x104 | 1x | 2/2 |
| Pseudomonas aeruginosa | UCLA P20 | 4x104 | 0.8x | 2/2 |
| Serratia marcescens | ATCC 8100 | 3x105 | 3x | 2/2 |
| Serratia marcescens | ATCC 13880 | 3x105 | 3x | 2/2 |
| Serratia marcescens | ATCC 14756 | 3x105 | 3x | 2/2 |
| Serratia marcescens | ATCC 15365 | 3x105 | 3x | 2/2 |
| Serratia marcescens | ATCC 27117 f | 2x105 | 2x | 1/2 |
| | | 3x105 | 3x | 1/2 |
| | | 5x105 | 5x | 1/2 |
| | | 1x106 | 10x | 0/2 |
| Serratia marcescens | ATCC 43861 | 2x105 | 2x | 2/2 |
| Serratia marcescens ssp. sakuensis | DSM-17174 | 3x105 | 3x | 2/2 |
| Staphylococcus aureus | IDEXX VB962455 | 8x106 | 1.6x | 2/2 |
| Staphylococcus aureus | IDEXX VB9981353 | 8x106 | 1.6x | 2/2 |
| Staphylococcus aureus | IDEXX VB969039 | 8x106 | 1.6x | 2/2 |
| Staphylococcus aureus | ATCC BAA-2312 | 8x106 | 1.6x | 2/2 |
| Staphylococcus aureus | NCTC 12493 | 6x106 | 1.2x | 2/2 |
| Staphylococcus aureus | ATCC 33591 | 6x106 | 1.2x | 2/2 |
| Staphylococcus aureus | DSM-17091 | 6x106 | 1.2x | 2/2 |
| Staphylococcus aureus | ATCC 12600 | 8x106 | 1.6x | 2/2 |
| Staphylococcus aureus | ATCC 29213 | 8x106 | 1.6x | 2/2 |
| Staphylococcus aureus | ATCC 43300 | 1.5x107 | 3x | 2/2 |
| Staphylococcus aureus | RKI 07-03165 | 8x106 | 1.6x | 2/2 |
| Staphylococcus aureus | RKI 01-00694 | 8x106 | 1.6x | 2/2 |
| Staphylococcus aureus | RKI 09-00187 | 8x106 | 1.6x | 2/2 |
| Staphylococcus aureus | RKI 08-02492 | 8x106 | 1.6x | 2/2 |
| Stenotrophomonas maltophilia | ATCC 13636 | 5x104 | 1.7x | 2/2 |
| | | 2x104 | 0.7x | 2/2 |
| | | 3x104 | 1x | 1/2 |
| Stenotrophomonas maltophilia | ATCC 13637 g | 5x104 | 1.7x | 5/6 |
| | | 1x105 | 3.3x | 1/2 |
| | | 2x105 | 6.7x | 1/2 |
| Stenotrophomonas maltophilia | ATCC 17666 | 2x104 | 0.7x | 2/2 |
| Stenotrophomonas maltophilia | ATCC 49130 | 2x104 | 0.7x | 2/2 |
| Stenotrophomonas maltophilia | DSM-50173 g<br>[ATCC 17444] | 2x104 | 0.7x | 1/4 |
| | | 3x104 | 1x | 0/1 |
| | | 5x104 | 1.7x | 3/4 |
| |…