Unyvero LRT BAL Application

K191967 · Curetis GmbH · QBH · Dec 20, 2019 · Microbiology

Device Facts

• Record ID: K191967
• Device Name: Unyvero LRT BAL Application
• Applicant: Curetis GmbH
• Product Code: QBH · Microbiology
• Decision Date: Dec 20, 2019
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3985
• Device Class: Class 2

Indications for Use

The Unyvero LRT BAL Application is a qualitative nucleic acid multiplex test intended for the simultaneous detection and identification of nucleic acid sequences from the following microorganisms (N = 20) and antibiotic resistance markers (N = 10) in bronchoalveolar lavage (BAL)-like specimens (BAL or mini-BAL) from adult hospitalized patients with suspected lower respiratory tract infections. The Unyvero LRT BAL 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 BAL specimens may contain colonizing microorganisms, detection of Unyvero LRT BAL microbial targets does not indicate that the microorganism is the disease. Unyvero positive results do not rule out co-infection with other microorganisms. 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 antibiotic resistance markers cannot be definitively linked to specific microorganisms, and may be present in organisms that are not detected by the Unyvero LRT BAL Application. Microbiology cultures of BALs should be performed to obtain isolates for species identification and antimicrobial susceptibility testing and to identify potential microorganisms not targeted by the Unyvero LRT BAL Application.

Device Story

The Unyvero LRT BAL Application is a multiplex PCR-based diagnostic system for identifying 20 lower respiratory pathogens and 10 antibiotic resistance markers directly from BAL or mini-BAL specimens. The system comprises the Unyvero L4 Lysator, A50 Analyzer, and C8 Cockpit. The workflow involves manual sample loading into a single-use cartridge, followed by automated DNA purification, eight parallel multiplex endpoint PCR reactions, and array-based hybridization. Fluorescence imaging of the arrays detects specific amplicons. The device is operated by laboratory personnel in a clinical setting. Results are displayed on the C8 Cockpit, providing qualitative detection of targets. The output aids clinicians in diagnosing lower respiratory tract infections and identifying potential antibiotic resistance, though results must be interpreted alongside clinical findings and traditional culture methods.

Clinical Evidence

Clinical performance was established via a prospective study (N=1,016) and an archived study (N=392). Prospective study PPA/NPA for typical microorganisms compared to SoC culture ranged widely (e.g., P. aeruginosa PPA 95.8%, NPA 95.4%; S. aureus PPA 88.7%, NPA 95.7%). A composite comparator (SoC culture + molecular PCR/sequencing) was also used. Antibiotic resistance marker performance was evaluated against molecular reference assays. Contrived samples supplemented data for rare analytes. Bench testing confirmed LoD, inclusivity, and exclusivity.

Technological Characteristics

The system uses multiplex endpoint PCR and array hybridization. Materials include a single-use cartridge containing DNA isolation reagents, primers, and hybridization buffers. The system is networked via Ethernet. Software manages the workflow across the Lysator, Analyzer, and Cockpit. The process is automated, requiring minimal manual intervention after sample loading. No specific ASTM standards for materials are cited; technical performance is defined by analytical sensitivity (LoD) and specificity.

Indications for Use

Indicated for adult hospitalized patients with suspected lower respiratory tract infections. Used as an aid in diagnosis of lower respiratory tract infection in patients with signs and symptoms. Not for definitive linkage of resistance markers to specific organisms; does not rule out co-infection; negative results do not preclude infection.

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.

Predicate Devices

• Unyvero LRT Application (DEN170047)

Submission Summary (Full Text)

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December 20, 2019

Curetis GmbH % Gail Radcliffe President Radcliffe Consulting, Inc. 231 Fairbanks Street West Boylston, Massachusetts 01583

Re: K191967

Trade/Device Name: Unyvero LRT BAL Application Regulation Number: 21 CFR 866.3985 Regulation Name: Device To Detect And Identify Microorganisms And Associated Resistance Marker Nucleic Acids Directly In Respiratory Specimens Regulatory Class: Class II Product Code: OBH Dated: July 22, 2019 Received: July 23, 2019

Dear Gail Radcliffe:

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

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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 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.

Kristian Roth, Ph.D. Branch Chief Bacterial Multiples and Medical Countermeasures Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health 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) K191967

Device Name Unyvero LRT BAL Application

#### Indications for Use (Describe)

The Unyvero LRT BAL Application is a qualitative nucleic acid multiplex test intended for the simultaneous detection and identification of nucleic acid sequences from the following microorganisms (N = 20) and antibiotic resistance markers (N = 10) in bronchoalveolar lavage (BAL)-like specimens (BAL or mini-BAL) from adult hospitalized patients with suspected lower respiratory tract infections.

[continued on page 2]

| Type of Use (Select one or both, as applicable) | |
|-------------------------------------------------|---------------------------------------------|
| { Prescription Use (Part 21 CFR 801 Subpart D) | Over-The-Counter Use (21 CFR 801 Subpart C) |

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[continued from Form FDA 3881, page 1]

| 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 complexc | 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 pneumoniaed | 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 | - |
| Pneumocystis jirovecii | - |
| 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 | - |

3 Acinetobacter spp. includes: A. baumannii, A. calcoaceticus, A. junii, A. nosocomialis, A. parvus, A. pitiii (detected by LRT BAL Application) and A. ursingii (not detected by LRT BAL Application).

b ctx-M1 subgroup.

° Enterobacter cloacae complex includes: E. chundaensis, E. cloacae, E. hormaechei (incl. ssp. xiang(angensis), E. kobei, E. ludwigii, E. roggenkampii, E. sichuandensis (not yet recognized as member of the E. cloacae complex).

d Klebsiella pneumoniae includes two variants: K. pneumoniae (variant 1), and K. quasipneumoniae (variant 2).

e Proteus spp. includes P. cibarius, P. hauseri, P. mirabilis, P. penneri and P. vulgaris.

The Unyvero LRT BAL 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 BAL specimens may contain colonizing microorganisms, detection of Unyvero LRT BAL microbial targets does not indicate that the microorganism is the disease. Unyvero positive results do not rule out co-infection with other microorganisms. 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 antibiotic resistance markers cannot be definitively linked to specific microorganisms, and may be present in organisms that are not detected by the Unyvero LRT BAL Application.

Microbiology cultures of BALs should be performed to obtain isolates for species identification and antimicrobial susceptibility testing and to identify potential microorganisms not targeted by the Unyvero LRT BAL Application.

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# 510(k) SUMMARY - LRT BAL Application (K191967)

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92. The assigned 510(k) number is K191967.

| <b>807.92 (a)(1):</b> | <b>Name:</b> | Curetis GmbH |
|-----------------------|-----------------|---------------------------------------------------------|
| | <b>Address:</b> | Max-Eyth-Strasse 42<br>Holzgerlingen, Germany 71088 |
| | <b>Phone:</b> | [phone] |
| | <b>Email:</b> | [e-mail] |
| | <b>Contact:</b> | Karsten Mueller, Head of Quality and Regulatory Affairs |

# 807.92 (a)(2): Device name - trade name and common name, and classification

# Trade name:

Unyvero LRT BAL Application

Common Name: Detection and identification of microorganisms and associated resistance marker nucleic acids directly in respiratory specimens

Classification / Product Code: 21 CFR Part 866.3985 / QBH

# 807.92 (a)(3): Identification of the legally marketed predicate devices

The Unyvero LRT BAL Application is substantially equivalent to its predecessor test system, the Unyvero LRT Application (Curetis, GmbH, Germany), granted de novo status under DEN170047 on April 3rd, 2018.

# 807.92 (a)(4): Device Description

The Unyvero LRT BAL Application automates and integrates DNA purification and eight parallel multiplex endpoint PCR reactions. It provides qualitative detection of nucleic acids from multiple lower respiratory pathogens using hybridization on PCR chamber arrays in a single use cartridge from a single bronchoalveolar lavage (BAL)-like specimen (BAL or mini-BAL).

The Unyvero LRT BAL Application identifies 20 microorganisms and 10 antibiotic resistance markers as listed in the Intended Use Statement, below.

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# Overview

The Unyvero LRT BAL Application uses a multiplex PCR approach following array hybridization which targets 30 individual analytes (microorganisms (N = 20) and antibiotic resistance markers (N = 10) divided into eight separate PCR reactions that are performed in individual reaction chambers simultaneously on a Unyvero LRT BAL Application cartridge. Multiplex compositions are designed to avoid any expected common occurrence of certain analytes within the same multiplex to largely reduce competitive PCR inhibition. Individual analyte assays of the Unyvero LRT BAL panel are designed to exhibit low or absent cross-reactivity with the relevant bronchoalveolar lavage (BAL)like specimens (BAL or mini-BAL) sample matrix or 'close neighbor' strains. Array oligonucleotides are designed for similar hybridization and melting temperatures (approx. 65 - 80 ℃, varying by amplicon). Hybridization and melting temperatures are used to exclude non-specific hybridization signals for improved signal specificity.

# Instrument. Cartridge, and Other Consumables

The instrumentation consists of one (or more) Unyvero L4 Lysator, one (or more) Unyvero A50 Analyzer, a Unyvero C8 Cockpit, and four single-use consumables: the Unyvero LRT BAL Cartridge, the Unyvero Sample Tube, Sample Tube Cap and the Unyvero Master Mix. A Unyvero Sample Tube Holder is supplied as accessory to simplify the sample filling step.

- Unyvero LRT BAL Cartridge contains DNA isolation and purification reagents, . primers, hybridization and wash buffers, and oligonucleotides for detection.
- Unyvero T1 Sample Tube and Transport Cap contains glass beads and buffers to lyse ● microorganisms and liquefy the sample.
- . Unyvero T1 Sample Tube Cap seals the Unyvero Sample Tube and contains Proteinase K and a synthetic control gene for process monitoring.
- Unyvero M1 Master Mix Tube contains reagents for DNA amplification. ●

# Controls

An internal control (a synthetic gene without any homology to known sequences) is processed in every chamber in order to verify the DNA purification, array hybridization, and detection.

Other than the built-in controls, no external materials are supplied with Unyvero LRT BAL Application devices and consumables. Good laboratory practice recommends running external positive and negative controls using samples cultured in the laboratory.

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## Reagents

No additional reagents are required to perform the Unyvero LRT BAL Application; all reagents are supplied within the cartridge or within the other consumables with the exception of the polymerase Master Mix, which is provided separately (frozen).

## Assav Procedures

How to perform a Unyvero LRT BAL test:

- 1. Remove the Unyvero Sample Tube from its packaging and slide it in the Unyvero Sample Tube Holder in the upright position with the barcoded end at the bottom.
- 2. Remove the Transport Cap from the Sample Tube by pulling it upward.
- 3. Pipette 180 uL of vortexed patient specimen into the Sample Tube and close it using the Unyvero Cap provided in the LRT BAL kit: align the small nodules on the neck of the Sample Tube with the openings on the Cap and press down to lock in place.
- 4. Scan the Sample Tube and place it into the Lysator. Close the Lysator lid to start the Lysator.
- 5. Remove Master Mix from freezer and thaw at room temperature (15 ℃ 25 ℃) for approximately 30 minutes.
- 6. When lysis is complete, remove the Sample Tube from the Lysator and place it into the labeled position on the left-hand side of the Unyvero LRT BAL Cartridge.
- 7. Place the thawed Master Mix into the labeled position on the right-hand side of the Cartridge.
- 8. Scan the Cartridge on the Cockpit and insert it into the position indicated on the Analyzer. The software provides on-screen instructions to start the test.
- 9. View results after completion of the run.

During the automated analysis (Step 8), which is entirely controlled by the A50 Analyzer, the sample is mixed with ethanol and then transferred onto the DNA purification column, where buffers purify and elute the DNA. Eluted 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, consisting of one labeled and one non-labeled primer for the respective multiplex endpoint PCR. After specific amplification, PCR products are hybridized to the corresponding array probes. Each array has been manufactured with probes corresponding to the amplicons for the targeted microorganism sequences described above. A total of up to 49 spots per array allows for redundant detection with at least four spots per analyte, as well as spots for intensity calibration, and orientation markers for the image processing software. Binding of amplicons to specific probes is detected by analyzing fluorescence images of the arrays. Result data are displayed on the C8 Cockpit for visualization, printout, temporary storage and electronic data export.

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A test run is completed after 4 to 5 hrs, and results for panel microorganisms and corresponding antibiotic resistance markers are displayed on the Cockpit screen. Four screens are provided:

- . The Result Summary screen provides a quick overview of all detected LRT BAL panel microorganisms, together with all detected corresponding antibiotic resistance markers.
- . The Microorganisms screen provides a list of all panel microorganisms grouped in Grampositive bacteria, non-fermenting bacteria, Enterobacteriaceae and other microorganisms together with the analysis result (reported as detected, not detected or invalid).
- The Result Details screen provides a list of all analyzed microorganisms and the ● corresponding antibiotic resistance markers together with the analysis result (reported as detected, not detected, invalid or NA).
- Test Details screen showing user name, lot numbers of used consumables, expiration dates of ● the consumables and start and stop times and dates of the test.

Results can be reviewed on the cockpit or, optionally, be printed out. All results are saved in a database on the Unyvero Cockpit for later review and printing.

## Software

The Unyvero software is designed to:

- Manage analysis workflow (Cockpit) ●
- Carry out sample lysis (Lysator) ●
- Execute the analysis and generate the analytical result (Analyzer) .
- Manage communication among units (Cockpit, Lysator, Analyzer) ●
- Monitor internal mechanical / electrical actuators (Lysator, Analyzer) ●
- Present analysis results (Cockpit) .
- Store analysis results (Cockpit) ●

Each device (Cockpit, Lysator. Analyzer) is a subsystem within the overall system, and each consists of hardware and software components. The different devices are interconnected by an Ethernet based communication interface, and system functionality is provided by the interaction of all three device types. Only the Cockpit presents a rich user interface and allows interaction with the operator. The Lysator and Analyzer units include a simple display for showing device status. Optional HIS/LIS connectivity allows transferring results to a hospital or laboratory information system.

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# 807.92 (a)(5): Intended Use

The Unyvero LRT BAL Application is a qualitative nucleic acid multiplex test intended for the simultaneous detection and identification of nucleic acid sequences from the following microorganisms (N = 20) and antibiotic resistance markers (N = 10) in bronchoalveolar lavage (BAL)like specimens (BAL or mini-BAL) from adult hospitalized patients with suspected lower respiratory tract infections.

| Microorganism | Associated Antibiotic Resistance Marker(s) |
|-------------------------------|-----------------------------------------------|
| 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 complexc | 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 pneumoniaed | 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 | - |
| Pneumocystis jirovecii | - |
| 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 | - |

® Acinetobacter spp. includes: A. baumannii, A. haemolyticus, A. junii, A. hvoffii, A. nosocomialis, A. purvus, A. pitti (detected by LRT BAL Application), and A. ursingii (not detected by LRT BAL Application).

b ctx-M1 subgroup.

§ Enterobacter cloacae complex includes: E. chengduensis, E. chundaensis, E. cloacae, E. hormaechei (incl. ssp. xiangfangensis), E. kobei, E. ludwigii, E. sichuanensis and E. bugandensis (not yet recognized as member of the E. cloacae complex).

e Proteus spp. includes: P. cibarius, P. hauseri, P. mirabilis, P. penneri, and P. vulgaris.

The Unyvero LRT BAL 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 BAL specimens may contain colonizing microorganisms, detection of Unyvero LRT BAL

510(k) Summary - LRT BAL Application K191967 Rev 3.0

d Klebsiella pneumoniae includes two variants: K. pneumoniae (variant 1), and K. quasipneumoniae (variant 2).

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microbial targets does not indicate that the microorganism is the cause of the disease. Unyvero positive results do not rule out co-infection with other microorganisms. 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 antibiotic resistance markers cannot be definitively linked to specific microorganisms, and may be present in organisms that are not detected by the Unyvero LRT BAL Application.

Microbiology cultures of BALs should be performed to obtain isolates for species identification and antimicrobial susceptibility testing and to identify potential microorganisms not targeted by the Unyvero LRT BAL Application.

# 807.92 (a)(6): Technological Similarities and Differences to the Predicate

| Element | New Device:<br>Curetis Unyvero LRT BAL Application | Predicate:<br>Curetis Unyvero LRT Application<br>(DEN170047) |
|-----------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Specimen Type | Bronchoalveolar lavage (BAL)-like<br>specimens from adult hospitalized patients<br>with suspected lower respiratory tract<br>infections | Endotracheal aspirate specimens from adult<br>hospitalized patients with suspected lower<br>respiratory tract infections |
| Organisms<br>Detected | Bacteria:<br>Acinetobacter spp.<br>Citrobacter freundii<br>Enterobacter cloacae complex<br>Escherichia coli<br>Haemophilus influenzae<br>Klebsiella oxytoca<br>Klebsiella pneumoniae<br>Klebsiella variicola<br>Moraxella catarrhalis<br>Morganella morganii<br>Proteus spp.<br>Pseudomonas aeruginosa<br>Serratia marcescens<br>Staphylococcus aureus<br>Stenotrophomonas maltophilia<br>Streptococcus pneumoniae<br>Atypical Bacteria:<br>Chlamydia pneumoniae<br>Legionella pneumophila<br>Mycoplasma pneumoniae<br>Fungus: Pneumocystis jirovecii<br>Antimicrobial Resistance Genes: | Bacteria:<br>Acinetobacter spp.<br>Citrobacter freundii<br>Enterobacter cloacae complex<br>Escherichia coli<br>Haemophilus influenzae<br>Klebsiella oxytoca<br>Klebsiella pneumoniae<br>Klebsiella variicola<br>Moraxella catarrhalis<br>Morganella morganii<br>Proteus spp.<br>Pseudomonas aeruginosa<br>Serratia marcescens<br>Staphylococcus aureus<br>Stenotrophomonas maltophilia<br>Streptococcus pneumoniae<br>Atypical Bacteria:<br>Chlamydia pneumoniae<br>Legionella pneumophila<br>Mycoplasma pneumoniae<br>Antimicrobial Resistance Genes: |

| | | Table 1: Comparison of the Unyvero LRT Application and the Unyvero LRT BAL Application |
|--|--|----------------------------------------------------------------------------------------|
| | | |

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| | ctx-M | ctx-M |
|-----------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------|
| | kpc | kpc |
| | mecA | mecA |
| | ndm | ndm |
| | oxa-23 | oxa-23 |
| | oxa-24 | oxa-24 |
| | oxa-48 | oxa-48, |
| | oxa-58 | oxa-58, |
| | tem | tem |
| | vim | vim |
| Analyte | DNA | DNA |
| Technological<br>Principles | Multiplex nucleic acid | Multiplex nucleic acid |
| Result Types | Qualitative for all analytes | Qualitative for all analytes |
| Instrumentation | Unyvero System | Unyvero System |
| Time to Result | About 4-5 hrs | About 4-5 hrs |
| Reagent Storage | Room temperature<br>(below -20 °C, Master Mix Tube only) | Room temperature<br>(below -20 °C, Master Mix Tube only) |
| Test<br>Interpretation | Automated test interpretation and report<br>generation. | Automated test interpretation and report<br>generation. |
| Controls | Control included in each test to monitor for<br>the presence of PCR inhibitors and enables<br>the system to detect failures in the testing<br>process. | Control included in each test to monitor for<br>the presence of PCR inhibitors and enables<br>the system to detect failures in the testing<br>process. |
| User Complexity | Moderate | Moderate |

# 807.92 (b)(1): Brief Description of Nonclinical Data

# Limits-of-Detection (LoDs)

Limits-of-Detection were determined for each LRT BAL panel analyte by serial dilutions of reference strains prepared in pooled negative native lavage specimens as test matrix. The lowest test concentration with a positivity rate of 95% or higher was determined as the LoD for each specific panel analyte (e.g., at least 19 of 20 positive results for at least 20 performed test replicates). Tables 2 and 3 summarize LoDs for all LRT BAL panel microorganisms or antibiotic resistance markers, respectively.

{11}------------------------------------------------

| | Reference<br>Strain ID | LoD<br>[CFU/mL] |
|-------------------------------------------------------|-------------------------------------------------------------|-----------------|
| Acinetobacter spp. | ATCC 19606<br>(A. baumannii) | 2.0 x 104 |
| Chlamydia pneumoniae | ATCC VR-2282 a | 3.2 x 102 |
| Citrobacter freundii | ATCC 8090 | 8.0 x 104 |
| Enterobacter cloacae complex | ATCC 13047<br>(E. cloacae) | 2.0 x 105 |
| Escherichia coli | ATCC 11775 | 2.0 x 104 |
| Haemophilus influenzae | ATCC 33391 | 2.0 x 104 |
| Klebsiella oxytoca | ATCC 13182 | 1.0 x 104 |
| Klebsiella pneumoniae (var. 1) | ATCC 13883 | 4.0 x 104 |
| Klebsiella quasipneumoniae<br>(K. pneumoniae, var. 2) | ATCC 700603 | 4.0 x 104 |
| Klebsiella variicola | ATCC BAA-830 | 2.0 x 104 |
| Legionella pneumophila | ATCC 33152 | 8.0 x 104 |
| Moraxella catarrhalis | ATCC 25238 | 1.5 x 105 |
| Morganella morganii | ATCC 25830 | 2.0 x 104 |
| Mycoplasma pneumoniae | ATCC 29085 b | 1.6 x 103 |
| Pneumocystis jirovecii | 36_0314 c | 5.0 x 105 |
| Proteus spp. | ATCC 29906<br>(P. mirabilis)<br>ATCC 29905<br>(P. vulgaris) | 5.0 x 103 |
| Pseudomonas aeruginosa | ATCC 10145 | 6.3 x 102 |
| Serratia marcescens | ATCC 13880 | 4.0 x 104 |
| Staphylococcus aureus | ATCC 12600 | 1.5 x 105 |
| Stenotrophomonas maltophilia | ATCC 13637 | 5.0 x 103 |
| Streptococcus pneumoniae | ATCC 49619 | 2.0 x 104 |

ª Cell supernatant (in IFU/mL).

b Bacterial suspension, quantified by qPCR (in copies/mL).

& Positive clinical specimen, quantified by qPCR (in copies/mL). LoD was determined using DNA extracted from a positive clinical specimen, and then confirmed by testing dilutions of this clinical specimen at the LoD concentration (20/20 positive results).

{12}------------------------------------------------

| | Reference<br>Strain ID | Host Microorganism | LoD<br>[CFU/mL] |
|--------|------------------------|-------------------------|-----------------|
| ctx-M | NCTC 13443 | Klebsiella pneumoniae | 1.0 x 104 |
| kpc | NCTC 13438 | Klebsiella pneumoniae | 4.0 x 104 |
| mecA | NCTC 12493 | Staphylococcus aureus | 4.0 x 105 |
| ndm | NCTC 13443 | Klebsiella pneumoniae | 2.0 x 104 |
| oxa-23 | NCTC 13301 | Acinetobacter baumannii | 1.0 x 107 |
| oxa-24 | NCTC 13302 | Acinetobacter baumannii | 5.0 x 103 |
| oxa-48 | NCTC 13442 | Klebsiella pneumoniae | 3.0 x 105 |
| oxa-58 | NCTC 13305 | Acinetobacter baumannii | 5.0 x 104 |
| tema | NCTC 13443 | Klebsiella pneumoniae | 2.0 x 104 |
| vim | NCTC 13437 | Pseudomonas aeruginosa | 2.0 x 104 |

Table 3: LoDs for LRT BAL panel antibiotic resistance markers.

ª Although the LRT BAL Application reports tem results only for H. influenzae as corresponding host microorganism, LoD was determined with a tem positive E. coli strain. Note that inclusivity testing was also successfully performed with tem positive H. influenzae strains.

# Inclusivity

### LRT BAL Microorganisms

Inclusivity wet testing was performed with reference strains for each microorganism analyte at concentrations near the LoD with contrived samples using pooled native negative lavage specimens as sample matrix (Table 4). Reference strains used for LoD determination are considered inclusive and were used as positive controls. Inclusivity was established near LoD (< 5x LoD) for all but one of the tested reference strains. For one reference strain of M. pneumoniae, tested as genomic DNA extract, inclusivity was demonstrated with a reduced analytical sensitivity at 5x LoD, although no primer or probe deviations to the corresponding LRT BAL assay were present.

| Reference Strain | Strain ID | Test Conc.<br>[CFU/ml] | x-fold LoD | # Pos./<br># Exp. |
|--------------------------------------------------------------------------------------------|-----------------------------|------------------------|------------|-------------------|
| analyte: Acinetobacter spp. | | | | |
| Acinetobacter baumannii<br>(pos. control/LoD ref. strain) | ATCC 19606 | 4.0 x 104 | 2x | 6/6 |
| Acinetobacter baumannii | NCTC 13305 | 4.0 x 104 | 2x | 2/2 |
| Acinetobacter baumannii | NCTC 13301 | 4.0 x 104 | 2x | 2/2 |
| Acinetobacter baumannii | NCTC 13302 | 4.0 x 104 | 2x | 2/2 |
| Acinetobacter baumannii | Micromyx 6148 | 4.0 x 104 | 2x | 2/2 |
| Acinetobacter baumannii | Micromyx 4410 | 4.0 x 104 | 2x | 2/2 |
| Acinetobacter baumannii | JMI 49755 | 4.0 x 104 | 2x | 2/2 |
| Acinetobacter baumannii | NRZ-00449 | 1.0 x 104 | 0.5x | 2/2 |
| Acinetobacter baumannii | UCLA A4 | 1.0 x 104 | 0.5x | 2/2 |
| Acinetobacter calcoaceticus | ATCC 23055 | 4.0 x 104 | 2x | 2/2 |
| Acinetobacter lwoffii | ATCC 15309 | 4.0 x 104 | 2x | 2/2 |
| Acinetobacter haemolyticus | ATCC 17906 | 4.0 x 104 | 2x | 2/2 |
| Chlamydia pneumoniae | ATCC VR-2282 | 6.4 x 102 a | 2x | 4/4 |
| Reference Strain | Strain ID | Test Conc. [CFU/ml] | x-fold LoD | # Pos./ # Exp. |
| (pos. control/LoD ref. strain) | | | | |
| Chlamydia pneumoniae | ATCC VR-1310 | $6.4 x 10^2 a$ | 2x | 2/2 |
| Chlamydia pneumoniae | ATCC 53592 | $1.0 x 10^3 a$ | 3x | 2/2 |
| Citrobacter freundii<br>(pos. control/LoD ref. strain) | ATCC 8090 | $1.6 x 10^5$ | 2x | 4/4 |
| Citrobacter freundii | ATCC 43864 | $1.6 x 10^5$ | 2x | 2/2 |
| Citrobacter freundii | NCTC 8581 | $1.6 x 10^5$ | 2x | 2/2 |
| Citrobacter freundii | NRZ-00452 | $1.6 x 10^5$ | 2x | 2/2 |
| Citrobacter freundii | UCLA C1 | $1.6 x 10^5$ | 2x | 2/2 |
| analyte: Enterobacter cloacae complex | | | | |
| Enterobacter cloacae<br>(pos. control/LoD ref. strain) | ATCC 13047 | $4.0 x 10^5$ | 2x | 7/7 |
| Enterobacter cloacae | ATCC 23355 | $4.0 x 10^5$ | 2x | 2/2 |
| Enterobacter cloacae | ATCC 49141 | $4.0 x 10^5$ | 2x | 2/2 |
| Enterobacter cloacae | ATCC BAA-2468 | $4.0 x 10^5$ | 2x | 2/2 |
| Enterobacter cloacae | JMI 46239 | $4.0 x 10^5$ | 2x | 2/2 |
| Enterobacter cloacae | NRZ-00239 | $4.0 x 10^5$ | 2x | 2/2 |
| Enterobacter cloacae ssp. dissolvens | ATCC 23373 | $4.0 x 10^5$ | 2x | 2/2 |
| Enterobacter hormaechei | ATCC 49162 | $4.0 x 10^5$ | 2x | 2/2 |
| Enterobacter asburiae | ATCC 35953 | $4.0 x 10^5$ | 2x | 2/2 |
| Escherichia coli<br>(pos. control/LoD ref. strain) | ATCC 11775 | $4.0 x 10^4$ | 2x | 4/4 |
| Escherichia coli | ATCC 25922 | $4.0 x 10^4$ | 2x | 2/2 |
| Escherichia coli | ATCC 35218 | $4.0 x 10^4$ | 2x | 4/4 |
| Escherichia coli | ATCC BAA-2523 | $4.0 x 10^4$ | 2x | 2/2 |
| Escherichia coli | NCTC 13351 | $4.0 x 10^4$ | 2x | 4/4 |
| Escherichia coli | NCTC 13476 | $4.0 x 10^4$ | 2x | 2/2 |
| Escherichia coli | JMI 50067 | $4.0 x 10^4$ | 2x | 2/2 |
| Haemophilus influenzae<br>(non-typeable/non-capsulated)<br>(pos. control/LoD ref. strain) | ATCC 33391 | $4.0 x 10^4$ | 2x | 3/3 |
| Haemophilus influenzae (serotype a) | ATCC 9006 | $4.0 x 10^4$ | 2x | 2/2 |
| Haemophilus influenzae (serotype c) | ATCC 9007 | $4.0 x 10^4$ | 2x | 2/2 |
| Haemophilus influenzae (serotype b) | ATCC 10211 | $4.0 x 10^4$ | 2x | 1/2 |
| | | $6.0 x 10^4$ | 3x | 2/2 |
| Haemophilus influenzae (serotype b) | ATCC 49247 | $4.0 x 10^4$ | 2x | 2/2 |
| Haemophilus influenzae (serotype b) | ATCC 49766 | $4.0 x 10^4$ | 2x | 2/2 |
| Klebsiella oxytoca<br>(pos. control/LoD ref. strain) | ATCC 13182 | $4.0 x 10^4$ | 4x | 3/3 |
| Klebsiella oxytoca | ATCC 43863 | $4.0 x 10^4$ | 4x | 2/2 |
| Klebsiella oxytoca | ATCC 8724 | $4.0 x 10^4$ | 4x | 2/2 |
| Klebsiella oxytoca | ATCC 49131 | $4.0 x 10^4$ | 4x | 2/2 |
| Klebsiella oxytoca | NCIMB 12819 | $4.0 x 10^4$ | 4x | 2/2 |
| Klebsiella oxytoca | NRZ-22060 | $4.0 x 10^4$ | 4x | 2/2 |
| Reference Strain | Strain ID | Test Conc.<br>[CFU/ml] | x-fold LoD | # Pos./<br># Exp. |
| Klebsiella pneumoniae, variant 1<br>(pos. control/LoD ref. strain) | ATCC 13883 | 8.0 x 104 | 2x | 4/4 |
| Klebsiella quasipneumoniae<br>(K. pneumoniae, variant 2)<br>(pos. control/LoD ref. strain) | ATCC 700603 | 8.0 x 104 | 2x | 3/3 |
| Klebsiella pneumoniae | NCTC 13439 | 8.0 x 104 | 2x | 2/2 |
| Klebsiella pneumoniae | NCTC 13438 | 8.0 x 104 | 2x | 2/2 |
| Klebsiella pneumoniae | NCTC 13442 | 8.0 x 104 | 2x | 2/2 |
| Klebsiella pneumoniae | NCTC 13443 | 2.0 x 104 | 0.5x | 3/3 |
| Klebsiella pneumoniae | Micromyx 4653 | 8.0 x 104 | 2x | 2/2 |
| Klebsiella pneumoniae | Micromyx 4676 | 8.0 x 104 | 2x | 2/2 |
| Klebsiella pneumoniae | JMI 49767 | 2.0 x 104 | 0.5x | 2/2 |
| Klebsiella pneumoniae | NRZ-00002 | 2.0 x 104 | 0.5x | 2/2 |
| Klebsiella pneumoniae | NRZ-00103 | 8.0 x 104 | 2x | 2/2 |
| Klebsiella variicola<br>(pos. control/LoD ref. strain) | ATCC BAA-830 | 4.0 x 104 | 2x | 3/3 |
| Klebsiella variicola | clinical strain 1 | 4.0 x 104 | 2x | 2/2 |
| Klebsiella variicola | clinical strain 2 | 4.0 x 104 | 2x | 2/2 |
| Klebsiella variicola | clinical strain 3 | 4.0 x 104 | 2x | 2/2 |
| Klebsiella variicola | clinical strain 4 | 4.0 x 104 | 2x | 2/2 |
| Legionella pneumophila (serotype 1)<br>(pos. control/LoD ref. strain) | ATCC 33152 | 1.6 x 105 | 2x | 7/7 |
| Legionella pneumophila (serotype 2) | ATCC 33154 | 1.6 x 105 | 2x | 2/2 |
| Legionella pneumophila (serotype 3) | ATCC 33155 | 1.6 x 105 | 2x | 2/2 |
| Legionella pneumophila (serotype 6) | ATCC 33215 | 1.6 x 105 | 2x | 2/2 |
| Legionella pneumophila (serotype 8) | ATCC 35096 | 1.6 x 105 | 2x | 2/2 |
| Legionella pneumophila (serotype 10) | ATCC 43283 | 1.6 x 105 | 2x | 2/2 |
| Legionella pneumophila | UCLA L1 | 1.6 x 105 | 2x | 2/2 |
| Legionella pneumophila | UCLA L5 | 1.6 x 105 | 2x | 2/2 |
| Legionella pneumophila | UCLA L6 | 1.6 x 105 | 2x | 2/2 |
| Moraxella catarrhalis<br>(pos. control/LoD ref. strain) | ATCC 25238 | 3.0 x 105 | 2x | 4/4 |
| Moraxella catarrhalis | ATCC 43617 | 3.0 x 105 | 2x | 2/2 |
| Moraxella catarrhalis | ATCC 8176 | 3.0 x 105 | 2x | 2/2 |
| Moraxella catarrhalis | ATCC 25240 | 3.0 x 105 | 2x | 2/2 |
| Moraxella catarrhalis | ATCC 23246 | 3.0 x 105 | 2x | 3/3 |
| Moraxella catarrhalis | ATCC 49143 | 3.0 x 105 | 2x | 2/2 |
| Morganella morganii<br>(pos. control/LoD ref. strain) | ATCC 25830 | 4.0 x 104 | 2x | 2/3 |
| Morganella morganii | ATCC 8019 | 4.0 x 104 | 2x | 2/2 |
| Morganella morganii | ATCC 25829 | 4.0 x 104 | 2x | 0/1 |
| | | 6.0 x 104 | 3x | 2/2 |
| Morganella morganii | DSM-46262 | 4.0 x 104 | 2x | 1/2 |
| | | 6.0 x 104 | 3x | 2/2 |
| Morganella morganii ssp. sibonii | ATCC 49948 | 4.0 x 104 | 2x | 2/2 |
| Mycoplasma pneumoniae (type 1)<br>(pos. control/LoD ref. strain) | ATCC 29085 | 3.2 x 103 b | 2x | 5/7 |
| Reference Strain | Strain ID | Test Conc.<br>[CFU/ml] | x-fold LoD | # Pos./<br># Exp. |
| Mycoplasma pneumoniae (type 1) | ATCC 29343 | 3.2 x 103 b | 2x | 2/2 |
| | | 3.2 x 103 b | 2x | 0/2 |
| | | 5.0 x 103 b | 3x | 1/2 |
| Mycoplasma pneumoniae (type 2) | ATCC 15531 c, d | 6.4 x 103 b | 4x | 1/2 |
| | | 7.0 x 103 b | 4.4x | 1/2 |
| | | 8.0 x 103 b | 5x | 2/2 |
| Mycoplasma pneumoniae (type 1) | ATCC 15293 | 3.2 x 103 b | 2x | 2/2 |
| Mycoplasma pneumoniae (type 2) | ATCC 49894 | 3.2 x 103 b | 2x | 2/2 |
| Pneumocystis jirovecii<br>(pos. control/LoD ref. strain) | pos. specimen 1 | 1.0 x 106 b | 2x | 1/1 |
| Pneumocystis jirovecii | pos. specimen 2 | 1.0 x 106 b | 2x | 2/2 |
| Pneumocystis jirovecii | pos. specimen 3 | 1.0 x 106 b | 2x | 2/2 |
| Pneumocystis jirovecii | pos. specimen 4 | 1.0 x 106 b | 2x | 2/2 |
| Pneumocystis jirovecii | pos. specimen 5 | 1.0 x 106 b | 2x | 2/2 |
| analyte: Proteus spp. | | | | |
| Proteus vulgaris<br>(pos. control/LoD ref. strain) | ATCC 29905 | 1.0 x 104 | 2x | 7/7 |
| Proteus mirabilis<br>(pos. control/LoD ref. strain) | ATCC 29906 | 1.0 x 104 | 2x | 2/2 |
| Proteus mirabilis | ATCC 12453 | 1.0 x 104 | 2x | 2/2 |
| Proteus mirabilis | ATCC 14153 | 1.0 x 104 | 2x | 2/2 |
| Proteus mirabilis | ATCC 25933 | 1.0 x 104 | 2x | 2/2 |
| Proteus vulgaris | ATCC 6380 | 1.0 x 104 | 2x | 2/2 |
| Proteus vulgaris | ATCC 8427 | 1.0 x 104 | 2x | 2/2 |
| Proteus hauseri | ATCC 700826 | 1.0 x 104 | 2x | 2/2 |
| Proteus penneri | ATCC 33519 | 1.0 x 104 | 2x | 2/2 |
| Pseudomonas aeruginosa<br>(pos. control/LoD ref. strain) | ATCC 10145 | 1.3 x 103 | 2x | 3/3 |
| Pseudomonas aeruginosa | ATCC 27853 | 1.3 x 103 | 2x | 2/2 |
| Pseudomonas aeruginosa | NCTC 13437 | 1.3 x 103 | 2x | 2/2 |
| Pseudomonas aeruginosa | Micromyx 2562 | 2.0 x 103 | 3x | 2/2 |
| Pseudomonas aeruginosa | NRZ-00196 | 1.3 x 103 | 2x | 2/2 |
| Pseudomonas aeruginosa | UCLA P20 | 1.3 x 103 | 2x | 2/2 |
| Serratia marcescens<br>(pos. control/LoD ref. strain) | ATCC 13880 | 8.0 x 104 | 2x | 3/3 |
| Serratia marcescens | ATCC 14756 | 8.0 x 104 | 2x | 2/2 |
| Serratia marcescens | ATCC 15365 | 8.0 x 104 | 2x | 2/2 |
| Serratia marcescens | ATCC 27117 | 8.0 x 104 | 2x | 2/2 |
| Serratia marcescens | ATCC 43861 | 8.0 x 104 | 2x | 2/2 |
| Serratia marcescens ssp. sakuensis | DSM-17174 | 8.0 x 104 | 2x | 2/2 |
| Staphylococcus aureus<br>(pos. control/LoD ref. strain) | ATCC 12600 | 3.0 x 105 | 2x | 7/7 |
| Staphylococcus aureus | ATCC BAA-2312 | 3.0 x 105 | 2x | 2/2 |
| Staphylococcus aureus | NCTC 12493 | 3.0 x 105 | 2x | 2/2 |
| Staphylococcus aureus | ATCC 33591 | 3.0 x 105 | 2x | 2/2 |
| Staphylococcus aureus | DSM-17091 | 3.0 x 105 | 2x | 2/2 |
| Reference Strain | Strain ID | Test Conc.<br>[CFU/ml] | x-fold LoD | # Pos./<br># Exp. |
| Staphylococcus aureus | ATCC 29213 | 3.0 x 105 | 2x | 2/2 |
| Staphylococcus aureus | ATCC 43300 | 3.0 x 105 | 2x | 2/2 |
| Stenotrophomonas maltophilia<br>(pos. control/LoD ref. strain) | ATCC 13637 | 1.0 x 104 | 2x | 4/4 |
| Stenotrophomonas maltophilia | ATCC 13636 | 1.0 x 104 | 2x | 0/2 |
| Stenotrophomonas maltophilia | | 1.5 x 104 | 3x | 2/2 |
| Stenotrophomonas maltophilia | ATCC 17666 | 1.0 x 104 | 2x | 2/2 |
| Stenotrophomonas maltophilia | ATCC 49130 | 1.0 x 104 | 2x | 2/2 |
| Stenotrophomonas maltophilia | DSM-50173 | 1.0 x 104 | 2x | 2/2 |
| Stenotrophomonas maltophilia | DSM-21874 e<br>[NCIMB 9528] | 1.0 x 104 | 2x | 0/2 |
| Stenotrophomonas maltophilia | | 1.5 x 104 | 3x | 0/2 |
| Stenotrophomonas maltophilia | | 2.0 x 104 | 4x | 2/2 |
| Streptococcus pneumoniae<br>serotype 19F<br>(pos. control/LoD ref. strain) | ATCC 49619 | 4.0 x 104 | 2x | 4/7 f |
| Streptococcus pneumoniae serotype 3 | ATCC 6303 | 4.0 x 104 | 2x | 2/2 |
| Streptococcus pneumoniae serotype 5 | ATCC 6305 | 4.0 x 104 | 2x | 2/2 |
| Streptococcus pneumoniae | ATCC 49150g | 4.0 x 104 | 2x | 0/2 |
| | | 8.0 x 104 | 4x | 2/2 |
| Streptococcus pneumoniae | ATCC 33400 h | 4.0 x 104 | 2x | 1/2 |
| | | 6.0 x 104 | 3x | 1/2 |
| | | 8.0 x 104 | 4x | 2/2 |
| Streptococcus pneumoniae serotype 2 | ATCC 27336 | 4.0 x 104 | 2x | 2/2 |
| Streptococcus pneumoniae serotype 1 | ATCC 6301 | 4.0 x 104 | 2x | 2/2 |
| Streptococcus pneumoniae serotype 9V | DSM-11865 | 4.0 x 104 | 2x | 2/2 |
| Streptococcus pneumoniae serotype 23F | DSM-11866 | 4.0 x 104 | 2x | 2/2 |
| | | 6.0 x 104 | 3x | 1/2 |
| | | 8.0 x 104 | 4x | 2/2 |
| Streptococcus pneumoniae serotype 6B | DSM-11867 | 4.0 x 104 | 2x | 2/2 |

#### Table 4: Inclusivity study results for LRT BAL panel microorganisms.

510(k) Summary - LRT BAL Application K191967 Rev 3.0

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

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

510(k) Summary – LRT BAL Application K191967 Rev 3.0

{15}------------------------------------------------

{16}------------------------------------------------

ª In IFU/mL for C. pneumoniae.

b In copies/mL for M. pneumoniae and P. jirovecii.

& Quantified DNA extract (reference material) from a commercial provider.

4 For M. pneumoniae ATCC 15531, a positivity rate of 2/2 was obtained at a 5x LoD concentration. Sequencing did not reveal any mismatches to primer and probe binding sites and the observed slightly is likely due to the different source material (DNA extract instead of a cell suspension).

e S. maltophilia strain DSM-21874 was positive only at a 4x LoD concentration. Sequencing did not reveal any mismatches to primer or probe sequences.

T S. pneumoniae LoD reference strain ATCC 46916 did not show consistently positive signals at 2x LoD. ATCC 46916 controls were repeated using a freshly prepared counted culture stock. This time, a positivity rate of 8/8 was obtained at a 2x LoD concentration, as expected.

& S. pneumoniae strain ATCC 49150 was only positive at 4x LoD. Sequencing did not reveal any mismatches to primer or probe sequences.

4 S. pneumoniae strain ATCC 33400 was only consistently positive at 4x LoD. Sequencing did not reveal any mismatches to primer or probe sequences.

{17}------------------------------------------------

To supplement inclusivity testing, in silico GenBank BLAST analyses of LRT BAL panel primer and probe sequences were performed for each LRT BAL panel microorganism (search performed July 2019) for all applicable strain entries. BLAST analyses identified strain entries for which detection by LRT BAL is predicted at LoD (match of relevant primer and probe sequences), only with reduced sensitivity (typically, single relevant mismatches of primer or probe sequences; detection likely at higher than LoD concentrations only), or is not predicted (multiple relevant mismatches in primer and probe sequences).

Table 5 lists microorganisms for which inclusivity was demonstrated by wet testing and are supported by in silico analysis (predicted at LoD or predicted with reduced sensitivity). Note that additional in silico analysis results are provided for strains that have not been wet tested. Such in silico results are provided as supplementary data only. Results are not intended to be a surrogate for wet testing and do not assure that specific strains will be detected.

NOTE: The performance of the Unyvero LRT BAL Application has not been established for those microorganism species that were evaluated by in silico analysis only.

Table 5: Inclusive LRT BAL panel microorganisms as demonstrated by inclusivity wet testing and/or as predicted by in silico analysis (at LoD concentration or with reduced sensitivity). For strains for which in silico analysis predicts higher than LoD concentrations for one or more applicable entries, numbers of GenBank entries are additionally listed.

| | Wet<br>Testing | In Silico:<br>at LoD | In Silico:<br>Reduced<br>Sensitivity | Comment |
|--------------------------------------|----------------|-----------------------|--------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Acinetobacter spp. | | | | |
| A. baumannii | X | X | - | |
| A. calcoaceticus | X | X | - | |
| A. lwoffii | X | X | - | |
| A. haemolyticus | X | X | - | |
| A. nosocomialis | - | X | - | |
| A. pittii | - | X | - | |
| A. junii | - | X | - | |
| A. parvus | - | X | - | |
| A. lactucae | - | X | - | |
| A. oleivorans | - | X | - |…