LIAISON PLEX Gram-Negative Blood Culture Assay

K243013 · Luminex Corporation · PEN · Apr 18, 2025 · Microbiology

Device Facts

Record IDK243013
Device NameLIAISON PLEX Gram-Negative Blood Culture Assay
ApplicantLuminex Corporation
Product CodePEN · Microbiology
Decision DateApr 18, 2025
DecisionSESE
Submission TypeTraditional
Regulation21 CFR 866.3365
Device ClassClass 2
AttributesReal-World Evidence

Real-World Evidence

SubmissionDeviceSponsorRWD SourcesRWE Use SummaryKey Tags
K243013 · Apr 18, 2025LIAISON PLEX Gram-Negative Blood Culture AssayLuminex CorporationRemnant, de-identified clinical blood culture specimens; Archived clinical specimens identified by standard-of-care testingThe clinical performance of the LIAISON PLEX BCN Assay was evaluated using prospective and retrospective clinical specimens to demonstrate substantial equivalence to the predicate device.Clinical performance study; Prospective clinical specimens; Retrospective clinical specimens; Standard of care

Clinical Evidence

Study DesignPopulationComparatorKey Endpoints
Prospective Clinical Study (Arm 1); Multi-site prospective clinical study using remnant, de-identified blood culture specimens; Follow-up/Duration: Not applicable; Study Period: March 2024 to June 2024Patients with positive blood culture samples containing gram-negative bacteria; Sample Size: 351 prospective specimens; Number of Sites: 4Culture followed by VITEK 2, PCR/BDS, or bothPositive percent agreement (PPA) and negative percent agreement (NPA) for target organisms and resistance markers
Retrospective Sample Evaluation (Arm 2); Retrospective study using pre-selected, archived clinical specimens; Follow-up/Duration: Not applicable; Study Period: September 2019 to May 2024Patients with positive blood culture samples; Sample Size: 231 pre-selected specimens; Number of Sites: 8Standard of Care (SoC) testing confirmed by VITEK 2 and/or PCR/BDSPositive percent agreement (PPA) and negative percent agreement (NPA)

Indications for Use

The LIAISON PLEX® Gram-Negative Blood Culture (BCN) Assay, performed using the automated, sample-to-result LIAISON PLEX® System, is a qualitative multiplexed in vitro diagnostic test for the simultaneous detection and identification of selected gram-negative pathogens and/or selected genetic determinants associated with antimicrobial resistance in positive blood culture bottles. LIAISON PLEX® BCN Assay is performed directly on blood culture media using blood culture bottles identified as positive by a continuous monitoring blood culture system which contain gram-negative bacteria as determined by Gram stain. The LIAISON PLEX® BCN Assay detects and identifies the following: Resistance Markers: CTX-M (blaCTX-M), IMP (blaIMP), KPC (blaKPC), NDM (blaNDM), OXA (blaOXA), VIM (blaVIM), MCR, SME (blaSME). Gram Negative Genera and Species: Enterobacteriaceae / Morganellaceae, Acinetobacter baumannii, Acinetobacter spp., Citrobacter spp., Enterobacter spp. (1), Escherichia coli (2), Haemophilus influenzae, Klebsiella oxytoca, Klebsiella pneumoniae, Klebsiella variicola, Morganella morganii, Neisseria meningitidis, Proteus spp., Pseudomonas aeruginosa, Pseudomonas spp., Salmonella spp., Serratia marcescens, Stenotrophomonas maltophilia. (1) Due to reclassification, Klebsiella aerogenes will be reported Enterobacter spp. (2) LIAISON PLEX® BCN Assay will not distinguish between Escherichia coli and Shigella spp. (S. dysenteriae, S. boydii, S. flexneri and S. sonnei). LIAISON PLEX® BCN Assay contains targets for the detection of genetic determinants associated with resistance to carbapenems (blaCTX-M, blaIMP, blaKPC, blaNDM, blaOXA48-like, blaVIM, blaSME) to aid in the identification of potentially antimicrobial-resistant organisms in positive blood culture samples. In addition, the panel includes an assay for the detection of the mobilized genetic determinant MCR, an emerging marker of public health importance. The antimicrobial resistance gene or marker detected may or may not be associated with the agent responsible for disease. Negative results for these select antimicrobial resistance gene and marker assays do not indicate susceptibility, as multiple mechanisms of resistance to β-lactams and colistin exist. LIAISON PLEX® BCN Assay is indicated for use in conjunction with other clinical and laboratory findings to aid in the diagnosis of bacterial bloodstream infections (BSI). LIAISON PLEX® BCN Assay is not intended to monitor treatment of these infections. Sub-culturing of positive blood cultures is necessary to recover organisms for antimicrobial susceptibility testing (AST), for identification of organisms not detected by LIAISON PLEX® BCN Assay, to detect mixed infections that may not be detected by LIAISON PLEX® BCN Assay, for association of antimicrobial resistance marker genes to a specific organism, or for epidemiological typing.

Device Story

The LIAISON PLEX® BCN Assay is an automated, sample-to-result, in vitro diagnostic test performed on the LIAISON PLEX® System. It uses a single-use disposable cartridge to process positive blood culture samples. The system performs mechanical and chemical cell lysis, magnetic bead-based nucleic acid isolation, and microarray-based hybridization. Target-specific capture DNA on a microarray binds extracted nucleic acids, followed by hybridization with mediator and gold nanoparticle probes. Silver enhancement of these probes allows for light scatter measurement to determine the presence or absence of specific gram-negative pathogens and resistance markers. The device is used in clinical laboratories by trained personnel. Results are provided to healthcare providers to aid in the diagnosis of bacterial bloodstream infections. The output helps clinicians identify potential antimicrobial-resistant organisms, though sub-culturing remains necessary for definitive AST and epidemiological typing. The device benefits patients by providing rapid identification of pathogens and resistance markers, potentially enabling earlier targeted therapy.

Clinical Evidence

Clinical performance was evaluated using 582 clinical specimens (351 prospective, 231 pre-selected) and 746 contrived specimens. The study compared the LIAISON PLEX® BCN Assay against a reference method algorithm (culture/VITEK 2/PCR/BDS). Combined prospective and pre-selected sensitivity/PPA ranged from 33.3% to 100% across targets, with specificity/NPA generally >99%. Contrived specimen sensitivity/PPA was 97.2%-100% and specificity/NPA was 99.6%-100%. The testing success rate was 99.7% for clinical and 99.9% for contrived samples.

Technological Characteristics

The device is a fully automated, bench-top system using a single-use disposable cartridge. It utilizes mechanical/chemical cell lysis, magnetic bead-based nucleic acid isolation, and microarray-based hybridization with gold nanoparticle probes and silver enhancement. It is a non-amplified, direct detection test. Connectivity is integrated into the LIAISON PLEX® System. It is designed for professional use in clinical laboratories.

Indications for Use

Indicated for use in patients with suspected bacterial bloodstream infections (BSI) to aid in the diagnosis by detecting gram-negative pathogens and antimicrobial resistance markers directly from positive blood culture bottles containing gram-negative bacteria (Gram stain confirmed). Not for treatment monitoring.

Regulatory Classification

Identification

A multiplex nucleic acid assay for identification of microorganisms and resistance markers from positive blood cultures is a qualitative in vitro device intended to simultaneously detect and identify microorganism nucleic acids from blood cultures that test positive by Gram stain or other microbiological stains. The device detects specific nucleic acid sequences for microorganism identification as well as for antimicrobial resistance. This device aids in the diagnosis of bloodstream infections when used in conjunction with other clinical and laboratory findings. However, the device does not replace traditional methods for culture and susceptibility testing.

Special Controls

In combination with the general controls of the FD&C Act, the Verigene® Gram Positive Blood Culture Nucleic Acid Test is subject to the following special controls: The special controls for the BC-GP Assay are contained in the guideline document entitled "Class II Special Controls Guideline: Multiplex Nucleic Acid Assay for Identification of Microorganisms and Resistance Markers from Positive Blood Cultures."

*Classification.* Class II (special controls). The special control for this device is FDA's guideline document entitled “Class II Special Controls Guideline: Multiplex Nucleic Acid Assay for Identification of Microorganisms and Resistance Markers from Positive Blood Cultures.” For availability of the guideline document, see § 866.1(e).

Predicate Devices

Submission Summary (Full Text)

{0} FDA U.S. FOOD & DRUG ADMINISTRATION # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION # DECISION SUMMARY ASSAY ONLY # I Background Information: A 510(k) Number K243013 B Applicant Luminex Corporation C Proprietary and Established Names LIAISON PLEX Gram-Negative Blood Culture Assay D Regulatory Information | Product Code(s) | Classification | Regulation Section | Panel | | --- | --- | --- | --- | | PEN | Class II | 21 CFR 866.3365 - Multiplex Nucleic Acid Assay For Identification Of Microorganisms And Resistance Markers From Positive Blood Cultures | MI - Microbiology | | NSU | Class II | 21 CFR 862.2570 - Instrumentation for clinical multiplex test systems | CH - Clinical Chemistry | # II Submission/Device Overview: A Purpose for Submission: To obtain substantial equivalence determination for the LIAISON PLEX Gram-Negative Blood Culture Assay B Measurand: Nucleic acid sequences from the following gram-negative bacteria and antibiotic resistance markers: Enterobacteriaceae / Morganellaceae, Acinetobacter baumannii, Acinetobacter spp., Citrobacter spp., Enterobacter spp., Escherichia coli, Haemophilus influenzae, Klebsiella Food and Drug Administration 10903 New Hampshire Avenue Silver Spring, MD 20993-0002 www.fda.gov {1} *oxytoca, Klebsiella pneumoniae, Klebsiella variicola, Morganella morganii, Neisseria meningitidis, Proteus* spp., *Pseudomonas aeruginosa, Pseudomonas* spp., *Salmonella* spp., *Serratia marcescens, Stenotrophomonas maltophilia*, CTX-M (*bla*CTX-M), IMP (*bla*IMP), KPC (*bla*KPC), NDM (*bla*NDM), OXA (*bla*OXA), VIM (*bla*VIM), MCR, and SME (*bla*SME). ### **C Type of Test:** A multiplexed nucleic acid test intended for use with the automated LIAISON PLEX instrument for the qualitative *in vitro* detection and identification of nucleic acid from gram-negative bacteria in a positive blood culture media sample that demonstrates the presence of gram-negative bacteria as determined by Gram stain. ### **III Intended Use/Indications for Use:** #### **A Intended Use(s):** See Indications for Use below. #### **B Indication(s) for Use:** The LIAISON PLEX Gram-Negative Blood Culture (BCN) Assay, performed using the automated, sample-to-result LIAISON PLEX System, is a qualitative multiplexed *in vitro* diagnostic test for the simultaneous detection and identification of selected gram-negative pathogens and/or selected genetic determinants associated with antimicrobial resistance in positive blood culture bottles. LIAISON PLEX BCN Assay is performed directly on blood culture media using blood culture bottles identified as positive by a continuous monitoring blood culture system which contain gram-negative bacteria as determined by Gram stain. The BCN Assay detects and identifies the following: Resistance Markers: CTX-M (*bla*CTX-M) IMP (*bla*IMP) KPC (*bla*KPC) NDM (*bla*NDM) OXA (*bla*OXA) VIM (*bla*VIM) MCR SME (*bla*SME) Gram Negative Genera and Species: *Enterobacteriaceae / Morganellaceae* *Acinetobacter baumannii* *Acinetobacter* spp. *Citrobacter* spp. *Enterobacter* spp. (1) *Escherichia coli* (2) *Haemophilus influenzae* *Klebsiella oxytoca* *Klebsiella pneumoniae* *Klebsiella variicola* K243013 - Page 2 of 51 {2} *Morganella morganii* *Neisseria meningitidis* *Proteus* spp. *Pseudomonas aeruginosa* *Pseudomonas* spp. *Salmonella* spp. *Serratia marcescens* *Stenotrophomonas maltophilia* (1) Due to reclassification, *Klebsiella aerogenes* will be reported Enterobacter spp. (2) LIAISON PLEX BCN Assay will not distinguish between *Escherichia coli* and *Shigella* spp. (*S. dysenteriae*, *S. boydii*, *S. flexneri* and *S. sonnei*). LIAISON PLEX BCN Assay contains targets for the detection of genetic determinants associated with resistance to carbapenems (*bla*CTX-M, *bla*IMP, *bla*KPC, *bla*NDM, *bla*OXA48-like, *bla*VIM, *bla*SME) to aid in the identification of potentially antimicrobial-resistant organisms in positive blood culture samples. In addition, the panel includes an assay for the detection of the mobilized genetic determinant MCR, an emerging marker of public health importance. The antimicrobial resistance gene or marker detected may or may not be associated with the agent responsible for disease. Negative results for these select antimicrobial resistance gene and marker assays do not indicate susceptibility, as multiple mechanisms of resistance to β-lactams and colistin exist. LIAISON PLEX BCN Assay is indicated for use in conjunction with other clinical and laboratory findings to aid in the diagnosis of bacterial bloodstream infections (BSI). LIAISON PLEX BCN Assay is not intended to monitor treatment of these infections. Sub-culturing of positive blood cultures is necessary to recover organisms for antimicrobial susceptibility testing (AST), for identification of organisms not detected by LIAISON PLEX BCN Assay, to detect mixed infections that may not be detected by LIAISON PLEX BCN Assay, for association of antimicrobial resistance marker genes to a specific organism, or for epidemiological typing. ### **C Special Conditions for Use Statement(s):** Rx - For Prescription Use Only ### **D Special Instrument Requirements:** LIAISON PLEX Instrument and Software version 1.4 ## **IV Device/System Characteristics:** ### **A Device Description:** The LIAISON PLEX Gram Negative Blood Culture (BCN) Assay is an automated test for the detection and identification of nucleic acids from gram-negative bacteria in a positive blood culture sample. The LIAISON PLEX BCN Assay is performed directly on blood culture media using blood culture bottles identified as positive by a continuous monitoring blood culture system, and which contain gram-negative bacteria, as determined by Gram stain. K243013 - Page 3 of 51 {3} The LIAISON PLEX BCN Assay is performed on the LIAISON PLEX System. The LIAISON PLEX System is a fully automated, benchtop device that performs sample preparation and microarray-based hybridization for the detection of target-specific nucleic acids. The LIAISON PLEX BCN Assay components required to perform the test include the following single-use, disposables: - LIAISON PLEX BCN Assay Cartridge - LIAISON PLEX BCN Assay Transfer Pipettes The test reagents are supplied in a single, disposable test cartridge. The LIAISON PLEX BCN Assay has 33 different reportable targets. Reporting of these targets is based on detection of one or more of the nucleic acid targets. Results can be: - reviewed by the user on the touchscreen; - exported to a USB flash drive; and, - printed, or submitted to a Laboratory Information System (LIS) for reporting. The LIAISON PLEX System automates the BCN Assay sample analysis through the following steps: 1) Sample Preparation, 2) Hybridization, and 3) Signal Analysis. The image analysis of the microarray provides light signal intensities from the target-specific capture spots, as well as the negative control, background, and imaging control spots. # B Principle of Operation: The assay workflow is as follows: 1. The user adds the sample into the BCN Assay Cartridge and closes the cartridge sample port. 2. The user creates a new order in the LIAISON PLEX System and scans the barcode ID or manually enters the sample ID. 3. The user scans the barcode with the hand-held barcode reader. 4. The user inserts the BCN Assay Cartridge into the open bay and then initiates the run. This is repeated, as needed, for up to six cartridges. 5. The system scans the barcode located on the top of the BCN Assay Cartridge and automatically performs the appropriate assay based on the unique barcode on the cartridge. The following steps take place within the closed cartridge: 1. Sample Preparation – The sample is mixed with reagents and nucleic acid extraction occurs via mechanical and chemical cell lysis and magnetic bead-based nucleic acid isolation. The sample then goes through a series of washes to capture and purify the nucleic acid. The nucleic acid is then transferred into the hybridization unit. 2. Hybridization – Extracted nucleic acids hybridize to specific capture DNA probes arrayed on a glass slide in a microarray format and the bound target DNA, in turn, hybridizes with mediator and gold-nanoparticle probes. An image of the hybridization unit is taken and the LIAISON PLEX Software performs data analysis. 3. Signal Analysis – Gold nanoparticle probes bound specifically to target-containing spots in the microarray are silver-enhanced, and light scatter from the spots is measured and further analyzed to determine the presence (Detected) or absence (Not Detected) of a target. K243013 - Page 4 of 51 {4} The system completes all necessary steps for the assay. No user intervention is necessary once the run is initiated. Once the run is complete, the user removes the cartridge(s) and disposes of them in the appropriate waste container. Following a series of quality control checks a final call is made for the Target: Detected if the Target signal is above threshold and Not Detected otherwise. Failure of any quality control checks results in a “No Call” result. ## V Substantial Equivalence Information: ### A Predicate Device Name(s): Verigene Gram Negative Blood Culture Nucleic Acid Test (BC-GN) ### B Predicate 510(k) Number(s): K132843 ### C Comparison with Predicate(s): | Device & Predicate Device(s): | K243013 (Candidate device) | K132843 (Predicate device) | | --- | --- | --- | | Device Trade Name | LIASION PLEX Gram Negative Blood Culture (BCN) Assay | Verigene Gram Negative Blood Culture Nucleic Acid Test (BC-GN) | | **General Device Characteristic Similarities** | | | | Intended Use/Indications For Use | The LIAISON PLEX Gram-Negative Blood Culture (BCN) Assay, performed using the automated, sample-to-result LIAISON PLEX System, is a qualitative multiplexed *in vitro* diagnostic test for the simultaneous detection and identification of selected gram-negative pathogens and/or selected genetic determinants associated with antimicrobial resistance in positive blood culture bottles. LIAISON PLEX BCN Assay is performed directly on blood culture media using blood culture bottles identified as positive by a continuous monitoring blood culture system which contain gram-negative bacteria as determined by Gram stain. The BCN Assay detects and identifies the following: Resistance Markers: CTX-M (*bla*CTX-M) IMP (*bla*IMP) KPC (*bla*KPC) NDM (*bla*NDM) OXA (*bla*OXA) VIM (*bla*VIM) MCR SME (*bla*SME) | The Verigene Gram Negative Blood Culture Nucleic Acid Test (BC-GN), performed using the sample-to-result Verigene System, is a qualitative multiplexed *in vitro* diagnostic test for the simultaneous detection and identification of selected gram-negative bacteria and resistance markers. BC-GN is performed directly on blood culture media using blood culture bottles identified as positive by a continuous monitoring blood culture system and which contain gram-negative bacteria as determined by gram stain. BC-GN detects and identifies the following: Gram Negative Genera and Species: *Acinetobacter* spp. *Citrobacter* spp. *Enterobacter* spp. *Proteus* spp. *Escherichia coli* (1) | K243013 - Page 5 of 51 {5} | | Gram Negative Genera and Species: *Enterobacteriaceae / Morganellaceae* *Acinetobacter baumannii* *Acinetobacter* spp. *Citrobacter* spp. *Enterobacter* spp. (1) *Escherichia coli* (2) *Haemophilus influenzae* *Klebsiella oxytoca* *Klebsiella pneumoniae* *Klebsiella variicola* *Morganella morganii* *Neisseria meningitidis* *Proteus* spp. *Pseudomonas aeruginosa* *Pseudomonas* spp. *Salmonella* spp. *Serratia marcescens* *Stenotrophomonas maltophilia* (1) Due to reclassification, *Klebsiella aerogenes* will be reported *Enterobacter* spp. (2) LIAISON PLEX BCN Assay will not distinguish between *Escherichia coli* and *Shigella* spp. (*S. dysenteriae*, *S. boydii*, *S. flexneri* and *S. sonnei*). LIAISON PLEX BCN Assay contains targets for the detection of genetic determinants associated with resistance to carbapenems (*bla*CTX-M, *bla*IMP, *bla*KPC, *bla*NDM, *bla*OXA48-like, *bla*VIM, *bla*SME) to aid in the identification of potentially antimicrobial-resistant organisms in positive blood culture samples. In addition, the panel includes an assay for the detection of the mobilized genetic determinant MCR, an emerging marker of public health importance. The antimicrobial resistance gene or marker detected may or may not be associated with the agent responsible for disease. Negative results for these select antimicrobial resistance gene and marker assays do not indicate susceptibility, as multiple mechanisms of resistance to β-lactams and colistin exist. LIAISON PLEX BCN Assay is indicated for use in conjunction with other clinical and laboratory findings to aid in the diagnosis of bacterial bloodstream infections (BSI). LIAISON PLEX BCN Assay is not intended to monitor treatment of these infections. Sub-culturing of positive blood cultures is necessary to recover organisms for antimicrobial susceptibility testing | *Klebsiella pneumoniae* *Klebsiella oxytoca* *Pseudomonas aeruginosa* (1) BC-GN will not distinguish *Escherichia coli* from *Shigella* spp. (*S. dysenteriae*, *S. flexneri*, *S. boydii*, and *S. sonnei*) Resistance Markers: CTX-M (*bla*CTX-M) IMP (*bla*IMP) KPC (*bla*KPC) NDM (*bla*NDM) OXA (*bla*OXA) VIM (*bla*VIM) BC-GN is indicated for use in conjunction with other clinical and laboratory findings to aid in the diagnosis of bacterial bloodstream infections; however, it is not used to monitor these infections. Sub-culturing of positive blood cultures is necessary to recover organisms for antimicrobial susceptibility testing (AST), for identification of organisms not detected by BC-GN, to detect mixed infections that may not be detected by BC-GN, for association of antimicrobial resistance marker genes to a specific organism, or for epidemiological typing. | | --- | --- | --- | K243013 - Page 6 of 51 {6} | | (AST), for identification of organisms not detected by LIAISON PLEX BCN Assay, to detect mixed infections that may not be detected by LIAISON PLEX BCN Assay, for association of antimicrobial resistance marker genes to a specific organism, or for epidemiological typing. | | | --- | --- | --- | | Measurand | Target DNA sequences of gram-negative bacteria and antimicrobial resistance markers | Same | | Specimen Type | Positive blood culture (PBC) samples that contain gram-negative bacteria | Same | | Test Principle | Qualitative, multiplexed nucleic acid hybridization test on a microarray format using capture and mediator oligonucleotides for gold nanoparticle probe-based endpoint detection | Same | | Sample Extraction | Chemical and mechanical lysis followed by a chaotrope-based nucleic acid isolation procedure | Same | | Quality Controls | Two internal processing controls (extraction and hybridization controls). External controls are recommended to be run. | Same | | Test Interpretation Output | Detected/Not Detected | Same | | **General Device Characteristic Differences** | | | | Organisms Detected | *Enterobacteriaceae / Morganellaceae* *Acinetobacter baumannii* *Acinetobacter* spp. *Citrobacter* spp. *Enterobacter* spp. *Escherichia coli* *Haemophilus influenzae* *Klebsiella oxytoca* *Klebsiella pneumoniae* *Klebsiella variicola* *Morganella morganii* *Neisseria meningitidis* *Proteus* spp. *Pseudomonas aeruginosa* *Pseudomonas* spp. | *Acinetobacter* spp. *Citrobacter* spp. *Enterobacter* spp. *Escherichia coli* *Klebsiella oxytoca* *Klebsiella pneumoniae* *Proteus* spp. *Pseudomonas aeruginosa* | K243013 - Page 7 of 51 {7} | | *Salmonella* spp. *Serratia marcescens* *Stenotrophomonas maltophilia* | | | --- | --- | --- | | Resistance Markers Detected | CTX-M (*bla*CTX-M) IMP (*bla*IMP) KPC (*bla*KPC) NDM (*bla*NDM) OXA (*bla*OXA) VIM (*bla*VIM) MCR SME (*bla*SME) | CTX-M (*bla*CTX-M) IMP (*bla*IMP) KPC (*bla*KPC) NDM (*bla*NDM) OXA (*bla*OXA) VIM (*bla*VIM) | | Storage Requirements | 15-30°C (for cartridges only) | 2-8°C (for cartridges only) | | Test Design | LIAISON PLEX BCN Assay Cartridge, LIAISON PLEX BCN Assay Transfer Pipettes | Verigene BC-GN Test Cartridge, Verigene BC-GN Extraction Tray (with Tip Holder Assembly), Verigene BC-GN Utility Kit | | Required Equipment | LIAISON PLEX System | Verigene System, Reader and Processor SP | | Time to Result | 2.5 hours | ~ 2 hours | ## VI Standards/Guidance Documents Referenced: | Document # | Title | | --- | --- | | Class II Special Controls Guideline | Multiplex Nucleic Acid Assay for Identification of Microorganisms and Resistance Markers from Positive Blood Cultures; Class II Special Controls Guideline for Industry and Food and Drug Administration Staff (Issued May 27, 2015) | | Guidance Document | Electronic Submission Template for Medical Device 510(k) Submissions; Guidance for Industry and Food and Drug Administration Staff (October 2023) | | Guidance Document | Content of Premarket Submissions for Device Software Functions; Guidance for Industry and Food and Drug Administration Staff (June 2023) | | Guidance Document | Cybersecurity in Medical Devices: Quality System Considerations and Content of Premarket Submissions; Guidance for Industry and Food and Drug Administration Staff (Issued September 27, 2023) | | CLSI EP07 3rd Edition | Interference Testing in Clinical Chemistry | | CLSI EP12 3rd Edition | Evaluation of Qualitative Binary Output Examination Performance | | CLSI EP17-A2 | Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures; Approved Guideline - Second Edition | | CLSI EP24-A2 | Assessment of the Diagnostic Accuracy of Laboratory Tests Using Receiver Operating Characteristic Curves; Approved Guideline - Second Edition | | CLSI EP25-A | Evaluation of Stability of In Vitro Diagnostic Reagents; Approved Guideline | | ANSI AAMI ISO 14971:2019 | Application of Risk Management to Medical Devices | | ISO 15223-1 Fourth edition 2021-07 | Medical devices - Symbols to be used with information to be supplied by the manufacturer - Part 1: General requirements | | ISO 18113-1:2011 | In vitro diagnostic medical devices - Information supplied by the manufacturer (labeling). Part 1: Terms, definition and general requirements. | K243013 - Page 8 of 51 {8} | ISO 18113-2:2011 | In vitro diagnostic medical devices - Information supplied by the manufacturer (labeling) Part 2: In vitro diagnostic reagents for professional use | | --- | --- | | ISO 18113-3:2011 | In vitro diagnostic medical devices - Information supplied by the manufacturer (labeling) Part 3: In vitro diagnostic instruments for professional use | | ISO 20916 | In vitro diagnostic medical devices - Clinical performance studies using specimens from human subjects - Good study practice. First edition. 2019. | | ISO 23640 | In vitro diagnostic medical devices - Evaluation of stability of in vitro diagnostic reagents | | ISO 3864-1 | Graphic Symbols - Safety Colors and Safety Signs - Part 1. Design Principles for Safety Signs and Safety Markings (2011) | | IEC 60601-1-2 Edition 4.0 | Medical Electrical Equipment - Part 1-2: General Requirements for Basic Safety and Essential Performance - Collateral Standard: Electromagnetic Disturbances - Requirements and Tests (2014-02) | | IEC 61010-1 Edition 3.1, Consolidated Version | Safety Requirements for Electrical Equipment for Measurement Control and Laboratory Use - Part 1: General Requirements, Including Corrigendum 1. (2017-01) | | IEC 61326-2-6 Edition 3.0 | Electrical Equipment for Measurement Control and Laboratory Use - EMC Requirements - Part 2-6: Particular Requirements - In Vitro Diagnostic (IVD) Medical Equipment (2010-10) | | IEC 62366-1 Edition 1.1 2020-06 CONSOLIDATED VERSION | Medical devices - Part 1: Application of usability engineering to medical devices | | ISO 62304:2006 | Medical device software - Software life cycle processes | | ISTA 3A | Packaged-Products for Parcel Delivery System Shipment 70 kg (150 lb) or Less. (2018) | | ISTA 7D | Temperature Test for Transport Packaging | ## VII Performance Characteristics (if/when applicable): ### Analytical Performance: #### 1. Precision/Reproducibility: Precision/reproducibility was conducted according to CLSI EP05-A3$^{1}$ and the Special Controls Guideline$^{2}$ to assess the performance of the LIAISON PLEX Gram-Negative Blood Culture (BCN) Assay. #### *Site-to-Site* A site-to-site precision study was conducted at three sites (2 external, one internal). A total of 10 blood culture samples were tested: six representative on-panel organisms (*Haemophilus influenzae*, *Neisseria meningitidis*, *Acinetobacter baumannii* + OXA, *Escherichia coli* + MCR, *Klebsiella pneumoniae* + KPC, and *Serratia marcescens* + SME) individually cultured to ring positivity and to eight hours after ring positivity, one negative sample contrived with an off-panel organism (*Staphylococcus aureus*), and one negative blood culture matrix (NBM) sample. Testing was conducted on one lot of the candidate test. Ninety (90) data $^{1}$ CLSI. Evaluation of Precision of Quantitative Measurement Procedures; Approved Guideline—Third Edition. CLSI document EP05-A3. Wayne, PA: Clinical and Laboratory Standards Institute; 2014. $^{2}$ FDA Class II Special Controls Guideline: Multiplex Nucleic Acid Assay for Identification of Microorganisms and Resistance Markers from Positive Blood Cultures; Guideline for Industry and Food and Drug Administration Staff (Issued May 27, 2015) K243013 - Page 9 of 51 {9} points were generated for each on-panel member and culture timepoint (0, 8 hrs post-positivity): 3 replicates per run × 2 operators per day × 5 non-consecutive days × 3 sites = 90 data points. Acceptance criteria are ≥ 95% agreement of positive and negative samples with expected results. Results are summarized in **Table 1** below. **Table 1. Summary Results of Site-to-Site Reproducibility** | Organism | Target Type | Agreement with Expected Results | | | | 95% C.I. | | | --- | --- | --- | --- | --- | --- | --- | --- | | | | Site 1 | Site 2 | Site 3 | Overall | Lower | Upper | | *A. baumannii* | Ring Positive | 100% (30/30) | 100% (30/30) | 100% (30/30) | 100% (90/90) | 95.9% | 100% | | | Ring Positive +8 Hours | 100% (30/30) | 100% (30/30) | 100% (30/30) | 100% (90/90) | 95.9% | 100% | | *K. pneumoniae* | Ring Positive | 100% (30/30) | 100% (31/31) | 100% (30/30) | 100% (91/91) | 95.9% | 100% | | | Ring Positive +8 Hours | 100% (30/30) | 100% (30/30) | 100% (30/30) | 100% (90/90) | 95.9% | 100% | | *H. influenzae* | Ring Positive | 100% (30/30) | 100% (30/30) | 100% (30/30) | 100% (90/90) | 95.9% | 100% | | | Ring Positive +8 Hours | 100% (30/30) | 100% (30/30) | 100% (30/30) | 100% (90/90) | 95.9% | 100% | | *N. meningitidis* | Ring Positive | 100% (30/30) | 100% (30/30) | 100% (30/30) | 100% (90/90) | 95.9% | 100% | | | Ring Positive +8 Hours | 100% (30/30) | 100% (30/30) | 100% (30/30) | 100% (90/90) | 95.9% | 100% | | *E. coli* | Ring Positive | 100% (30/30) | 100% (30/30) | 100% (30/30) | 100% (90/90) | 95.9% | 100% | | | Ring Positive +8 Hours | 100% (30/30) | 100% (30/30) | 100% (30/30) | 100% (90/90) | 95.9% | 100% | | *S. marcescens* | Ring Positive | 100% (30/30) | 100% (30/30) | 100% (30/30) | 100% (90/90) | 95.9% | 100% | | | Ring Positive +8 Hours | 100% (30/30) | 100% (30/30) | 100% (30/30) | 100% (90/90) | 95.9% | 100% | | *S. aureus* | Off-panel Negative | 98.3% (59/60) | 100% (60/60) | 100% (60/60) | 99.4% (179/180) | 96.9% | 99.9% | | Negative Blood Culture | Off-panel Negative | 100% (60/60) | 100% (60/60) | 100% (60/60) | 100% (180/180) | 97.9% | 100% | | **Resistance Markers** | | | | | | | | | OXA | Ring Positive | 100% (30/30) | 100% (30/30) | 100% (30/30) | 100% (90/90) | 95.9% | 100% | | | Ring Positive +8 Hours | 100% (30/30) | 100% (30/30) | 100% (30/30) | 100% (90/90) | 95.9% | 100% | | KPC | Ring Positive | 100% (30/30) | 100% (31/31) | 100% (30/30) | 100% (91/91) | 95.9% | 100% | | | Ring Positive +8 Hours | 100% (30/30) | 100% (30/30) | 100% (30/30) | 100% (90/90) | 95.9% | 100% | | MCR | Ring Positive | 100% (30/30) | 100% (30/30) | 100% (30/30) | 100% (90/90) | 95.9% | 100% | | | Ring Positive +8 Hours | 100% (30/30) | 100% (30/30) | 100% (30/30) | 100% (90/90) | 95.9% | 100% | | SME | Ring Positive | 100% (30/30) | 100% (30/30) | 100% (30/30) | 100% (90/90) | 95.9% | 100% | | | Ring Positive +8 Hours | 100% (30/30) | 100% (30/30) | 100% (30/30) | 100% (90/90) | 95.9% | 100% | *Lot-to-Lot* K243013 - Page 10 of 51 {10} A lot-to-lot precision study was conducted using one operator at one internal site. Testing was conducted over a minimum of five non-consecutive days across three lots of LIAISON PLEX BCN Assay cartridges and two LIAISON PLEX systems. Data from the cartridge lot for Operator 1 tested during site-to-site testing was leveraged for assessment of lot-to-lot precision. A total of 6 blood culture samples were tested: two representative on-panel organisms (*A. baumannii* + OXA and *K. pneumoniae* + KPC) individually cultured to ring positivity and to eight hours after ring positivity, one negative sample contrived with an off-panel organism (*Staphylococcus aureus*), and one negative blood culture matrix (NBM) sample. Ninety (90) data points were generated per lot: 3 replicates per run × 5 days × 6 panel members = 90 data points. Acceptance criteria are ≥ 95% agreement of positive and negative samples with expected results. Results are summarized in **Table 2** below. **Table 2. Summary Results of Lot-to-Lot Reproducibility** | Organism | Target Type | Agreement with Expected Results | | | | 95% C.I. | | | --- | --- | --- | --- | --- | --- | --- | --- | | | | Lot 1 | Lot 2 | Lot 3 | Overall | Lower | Upper | | *A. baumannii* | Ring Positive | 100% (15/15) | 100% (15/15) | 100% (15/15) | 100% (45/45) | 92.1% | 100% | | | Ring Positive +8 Hours | 100% (15/15) | 100% (15/15) | 100% (15/15) | 100% (45/45) | 92.1% | 100% | | *K. pneumoniae* | Ring Positive | 100% (15/15) | 100% (15/15) | 100% (15/15) | 100% (45/45) | 92.1% | 100% | | | Ring Positive +8 Hours | 100% (15/15) | 100% (15/15) | 100% (15/15) | 100% (45/45) | 92.1% | 100% | | *S. aureus* | Off-panel Negative | 100% (15/15) | 100% (15/15) | 100% (15/15) | 100% (45/45) | 92.1% | 100% | | Negative Blood Culture | Off-panel Negative | 100% (15/15) | 100% (15/15) | 100% (15/15) | 100% (45/45) | 92.1% | 100% | | **Resistance Markers** | | | | | | | | | OXA | Ring Positive | 100% (15/15) | 100% (15/15) | 100% (15/15) | 100% (45/45) | 92.1% | 100% | | | Ring Positive +8 Hours | 100% (15/15) | 100% (15/15) | 100% (15/15) | 100% (45/45) | 92.1% | 100% | | KPC | Ring Positive | 100% (15/15) | 100% (15/15) | 100% (15/15) | 100% (45/45) | 92.1% | 100% | | | Ring Positive +8 Hours | 100% (15/15) | 100% (15/15) | 100% (15/15) | 100% (45/45) | 92.1% | 100% | #### *Repeatability and Within-Laboratory Precision* Within-lab precision/repeatability was assessed using site-to-site precision study data. Results from site-to-site testing are stratified according to operator in **Table 3** below. **Table 3. Summary Results of Within-Lab/Repeatability Stratified by Operator** | Organism | Target Type | Agreement with Expected Results | | | --- | --- | --- | --- | | | | Site 1 | | | | | Operator 1 | Operator 2 | | *A. baumannii* | Ring Positive | 100% 15/15 | 100% 15/15 | | | Ring Positive +8 Hours | 100% 15/15 | 100% 15/15 | | *K. pneumoniae* | Ring Positive | 100% 15/15 | 100% 15/15 | | | Ring Positive +8 Hours | 100% 15/15 | 100% 15/15 | | *H. influenzae* | Ring Positive | 100% 15/15 | 100% 15/15 | | | Ring Positive +8 Hours | 100% 15/15 | 100% 15/15 | | *N. meningitidis* | Ring Positive | 100% 15/15 | 100% 15/15 | | | Ring Positive +8 Hours | 100% 15/15 | 100% 15/15 | K243013 - Page 11 of 51 {11} | Organism | Target Type | Agreement with Expected Results | | | --- | --- | --- | --- | | | | Site 1 | | | | | Operator 1 | Operator 2 | | *E. coli* | Ring Positive | 100% 15/15 | 100% 15/15 | | | Ring Positive +8 Hours | 100% 15/15 | 100% 15/15 | | *S. marcescens* | Ring Positive | 100% 15/15 | 100% 15/15 | | | Ring Positive +8 Hours | 100% 15/15 | 100% 15/15 | | *S. aureus* | Off-panel Negative | 100% 30/30 | 96.7% 29/30 | | Negative Blood Culture | Off-panel Negative | 100% 30/30 | 100% 30/30 | # 2. Linearity: This study is not applicable as the test device is a qualitative assay. # 3. Analytical Specificity/Interference: The LIAISON PLEX Gram-Negative Blood Culture (BCN) Assay was evaluated for analytical specificity/cross-reactivity and interference. # *Cross-Reactivity* The LIAISON PLEX BCN Assay was evaluated for microbial cross-reactivity. A total of 113 “off-panel” organisms, including those phylogenetically related to the on-panel organisms and organisms likely to be present in clinical blood culture samples but not detected by the LIAISON PLEX BCN Assay. Each potentially cross-reacting organism was spiked into negative blood matrix and tested in triplicate. All species were tested at clinically relevant concentrations or the highest achievable concentration. **Table 4** lists the microorganisms tested and the results of microbial cross-reactivity testing. **Table 4. Summary Results of Microorganisms Evaluated for Cross-Reactivity** | Organism | Positivity (%) | Organism | Positivity (%) | | --- | --- | --- | --- | | **Gram Positive** | | | | | *Abitrophia defective* | 0% | *Leuconostoc carnosum* | 0% | | *Aerococcus viridans* | 0% | *Leuconostoc mesenteroids* | 0% | | *Arcanobacterium bernardiae* | 0% | *Listeria grayi* | 0% | | *Arcanobacterium haemolyticum* | 0% | *Listeria innocua* | 0% | | *Bacillus cereus* | 0% | *Listeria ivanovii* | 0% | | *Bacillus licheniformis* | 0% | *Listeria welshimeri* | 0% | | *Bacillus subtilis* | 0% | *Micrococcus luteus* | 0% | | *Bacillus thuringiensis* | 0% | *Micrococcus lylae* | 0% | | *Corynebacterium amycolatum* | 0% | *Pediococcus acidilactici* | 0% | | *Corynebacterium diphtheriae* | 0% | *Pediococcus pentosaceus* | 0% | | *Corynebacterium pseudodiphtheriticum* | 0% | *Peptostreptococcus anaerobius* | 0% | | *Corynebacterium striatum* | 0% | *Planococcus citreus* | 0% | | *Cutibacterium acnes* | 0% | *Planococcus kocurii* | 0% | | *Cutibacterium avidum* | 0% | *Rothia dentocariosa* | 0% | | *Propionibacterium freudenreichii* | 0% | *Rothia (Stomatococcus) mucilaginosa* | 0% | | *Enterococcus avium* | 0% | *Staphylococcus capitis* | 0% | | *Enterococcus casseliflavis*, VRE, vanC | 0% | *Staphylococcus caprae* | 0% | | *Enterococcus dispar* | 0% | *Staphylococcus cohnii* | 0% | | *Enterococcus durans* | 0% | *Staphylococcus haemolyticus* | 0% | | *Enterococcus flavescens* | 0% | *Staphylococcus hominis* | 0% | | *Enterococcus gallinarum*, vanC | 0% | *Staphylococcus intermedius* | 0% | K243013 - Page 12 of 51 {12} | Organism | Positivity (%) | Organism | Positivity (%) | | --- | --- | --- | --- | | Enterococcus hirae | 0% | Staphylococcus lugdunensis | 0% | | Enterococcus mundtii | 0% | Streptococcus agalactiae | 0% | | Enterococcus raffinosus | 0% | Streptococcus anginosus | 0% | | Erysipelothrix rhusiopathiae | 0% | Streptococcus bovis | 0% | | Kocuria kristinae | 0% | Streptococcus constellatus | 0% | | Kytococcus sedentarius, Met R | 0% | Streptococcus equinus | 0% | | Lactobacillus acidophilus | 0% | Streptococcus mitis | 0% | | Lactobacillus crispatus | 0% | Streptococcus pneumoniae | 0% | | Lactobacillus rhamnosus | 0% | Streptococcus pyogenes | 0% | | Corynebacterium frankenforstense | 0% | | | | **Gram Negative** | | | | | Aggregatibacter aphrophilus | 0% | Fusobacterium nucleatum | 0% | | Bacteroides fragilis | 0% | Haemophilus haemolyticus | 0% | | Brevundimonas diminuta | 0% | Herbaspirillum huttiense | 0% | | Burkholderia cepacia | 0% | Kingella kingae | 0% | | Capnocytophaga ochracea | 0% | Moraxella catarrhalis | 0% | | Cardiobacterium hominis | 0% | Neisseria lactamica | 0% | | Comamonas testosteronae | 0% | Neisseria mucosa | 0% | | Delftia acidovorans | 0% | Neisseria sicca | 0% | | Eikenella corrodens | 0% | Parabacteroides distasonis | 0% | | Elizabethkingia meningoseptica | 0% | Pasteurella aerogenes | 0% | | Fusobacterium necrophorum | 0% | Prevotella bivia | 0% | | Aeromonas hydrophila | 0% | Klebsiella quasivariicola | 100% | | Haemophilus parainfluenzae | 0% | Providencia rustigiani | 100% | | Haemophilus parahaemolyticus | 33% | Providencia vermicola | 100% | | Enterobacter kobei | 100% | Pseudomonas paraeruginosa | 100% | | Haemophilus aegyptius | 100% | Serratia entomophila | 100% | | Klebsiella grimontii | 100% | Serratia nevei | 100% | | Klebsiella huaxiensis | 100% | Serratia odorifera | 100% | | Klebsiella michiganensis | 100% | Serratia quinivorans | 100% | | Klebsiella quasipneumoniae | 100% | Serratia ureilytica | 100% | | Lelliottia amnigena | 100% | | | | **Yeast/Fungi** | | | | | Aspergillus fumigatus | 0% | Candida tropicalis | 0% | | Candida albicans | 0% | Cryptococcus neoformans | 0% | | Candida famata | 0% | Kluyveromyces lactis | 0% | | Candida glabrata | 0% | Saccharomyces cerevisiae | 0% | | Candida krusei | 0% | Schizosaccharomyces pombe | 0% | | Candida parapsilosis | 0% | | | Of the off-panel organisms tested, 17 species generated positive results for Enterobacteriaceae / Morganellaceae, and of those 17, 11 had an additional positive result that was found to be cross-reactive. Table 5 lists the microorganisms shown to cross-react and the false target calls made on the LIAISON PLEX BCN Assay. The observed cross-reactivity is noted in the device labeling. Table 5. Summary Results of Cross-Reactive Organisms | Organism | Positivity (%) | Results | | | --- | --- | --- | --- | | Lelliottia amnigena | 100% | Enterobacteriaceae / Morganellaceae | Enterobacter spp. | K243013 - Page 13 of 51 {13} | Pseudomonas paraeruginosa | 100% | Pseudomonas spp. | Pseudomonas aeruginosa | | --- | --- | --- | --- | | Serratia nevei | 100% | Enterobacteriaceae / Morganellaceae | Serratia marcescens | | Serratia entomophila | 100% | Enterobacteriaceae / Morganellaceae | Serratia marcescens | | Serratia ureilytica | 100% | Enterobacteriaceae / Morganellaceae | Serratia marcescens | | Klebsiella quasivariicola | 100% | Enterobacteriaceae / Morganellaceae | Klebsiella pneumoniae | | Haemophilus aegyptius | 100% | None | Haemophilus influenzae | | Klebsiella michiganensis | 100% | Enterobacteriaceae / Morganellaceae | Klebsiella oxytoca | | Klebsiella grimontii | 100% | Enterobacteriaceae / Morganellaceae | Klebsiella oxytoca | | Klebsiella quasipneumoniae | 100% | Enterobacteriaceae / Morganellaceae | Klebsiella pneumoniae | | Haemophilus parahaemolyticus | 33% | None | Enterobacteriaceae / Morganellaceae | # In Silico Inclusivity/Exclusivity In silico analyses were conducted for potentially cross-reactive organisms that could not be obtained for wet testing to predict potential cross-reactivity of the assay oligos through a BLAST comparison of the oligo sequences to the GenBank nucleotide (nt) sequence database residing locally on company servers. To ensure that all potential cross-reactivity is assessed, this analysis was performed with all capture and mediator probes in the assay's oligo mix, including the internal control oligos. Table 6 lists the microorganisms for which sequences were subjected to in silico analysis to predict potential cross-reactivity. Table 6. Potential Cross-Reactive Organisms Tested in In Silico Analysis | On-Panel Organisms | Off-Panel Organisms | | | | | --- | --- | --- | --- | --- | | | Gram-Positive Bacteria | Gram-Negative Bacteria | Resistance Markers | Yeasts / Viruses / Parasites | | Acinetobacter baumannii | Actinomyces israelii | Actinobacillus hominis | AmpC | Aspergillus flavus | | Acinetobacter spp. | Actinomyces naeslundii | Actinobacillus ureae | CMY | Aspergillus fumigatus | | Citrobacter spp. | Actinomyces odontolyticus | Aeromonas caviae | mecA | Aspergillus niger | | Enterobacter spp. | Aerococcus sanguinicola | Aeromonas hydrophila | mecC | Aspergillus terreus | | Enterobacteriaceae | Aerococcus urinae | Aeromonas sobria | ompK36 | Blastomyces dermatitidis | | Escherichia coli / Shigella spp. | Aerococcus viridans | Aggregatibacter actinomycetemcomitans | RAHN | Candida albicans | | Haemophilus influenzae | Arcanobacterium bernardiae | Aggregatibacter aphrophilus | SHV | Candida auris | | Klebsiella oxytoca | Arcanobacterium haemolyticum | Bacteroides caccae | SPM | Candida dubliniensis | | Klebsiella pneumoniae | Arthrobacter psychrolactophilus | Bacteroides fragilis | TEM | Candida duobushaemulonii | | Klebsiella variicola | Bacillus spp. | Bacteroides ovatus | vanA | Candida famata | | Morganella morganii | Brochothrix thermosphacta | Bacteroides thetaiotaomicron | vanB | Candida glabrata | | Morganellaceae | Carnobacterium divergens | Bacteroides uniformis | vanC | Candida guilliermondii | | Neisseria meningitidis | Carnobacterium maltaromaticum | Bacteroides vulgatus | vanD | Candida haemulonii | K243013 - Page 14 of 51 {14} | On-Panel Organisms | Off-Panel Organisms | | | | | --- | --- | --- | --- | --- | | | Gram-Positive Bacteria | Gram-Negative Bacteria | Resistance Markers | Yeasts / Viruses / Parasites | | Proteus spp. | Cellulomonas turbata | Bacteroides xylanisolvens | vanM | Candida inconspicua | | Pseudomonas aeruginosa | Cellulosimicrobium cellulans | Bordetella bronchiseptica | | Candida kefyr | | Pseudomonas spp. | Clostridioides difficile | Bordetella parapertussis | | Candida krusei | | Salmonella spp. | Clostridium bifermentans | Bordetella pertussis | | Candida lipolytica | | Serratia marcescens | Clostridium clostridioforme | Brevundimonas diminuta | | Candida lusitaniae | | Stenotrophomonas maltophilia | Clostridium perfringens | Brevundimonas vesicularis | | Candida metapsilosis | | CTX-M | Clostridium ramosum (Thomasclavelia ramosa) | Burkholderia cepacia | | Candida multis-gemmis | | IMP | Clostridium septicum | Burkholderia mallei | | Candida nivariensis | | KPC | Clostridium tertium | Burkholderia multivorans | | Candida norvegensis | | MCR (MCR-1, MCR-2, MCR-3) | Clostridium tetani | Burkholderia pseudomallei | | Candida orthopsilosis | | NDM | Corynebacterium spp. | Campylobacter hominis | | Candida parapsilosis | | OXA (OXA-23, OXA-24/40/143, OXA-48, OXA-58) | Cutibacterium acnes | Capnocytophaga ochracea | | Candida sojae | | SME | Cutibacterium avidum | Cardiobacterium hominis | | Candida tropicalis | | VIM | Cutibacterium granulosum | Chlamydia trachomatis | | Candida viswanathii | | | Enterococcus avium | Chlamydophila pneumoniae | | Coccidioides immitis | | | Enterococcus casseliflavus | Chromobacterium violaceum | | Coccidioides posadasii | | | Enterococcus cecorum | Comamonas testosteroni | | Cryptococcus amylolentus | | | Enterococcus dispar | Delftia acidovorans | | Cryptococcus gattii | | | Enterococcus durans | Eikenella corrodens | | Cryptococcus neoformans | | | Enterococcus faecalis | Elizabethkingia meningoseptica | | Cryptococcus uniguttulatus | | | Enterococcus faecium | Fusobacterium necrophorum | | Cutaneotrichosporon curvatum | | | Enterococcus flavescens | Fusobacterium nucleatum | | Cyberlindnera fabianii | | | Enterococcus gallinarum | Haemophilus aegyptius | | Geotrichum capitatum (Magnusiomyces capitatus) | | | Enterococcus hirae | Haemophilus ducreyi | | Histoplasma capsulatum | | | Enterococcus mundtii | Haemophilus haemolyticus | | Kluyveromyces lactis | | | Enterococcus raffinosus | Haemophilus parahaemolyticus | | Kodamaea ohmeri | | | Erysipelothrix rhusiopathiae | Haemophilus parainfluenzae | | Lodderomyces elongisporus | | | Finegoldia magna | Haemophilus parasuis | | Magnusiomyces capitatus | | | Gemella haemolysans | Haemophilus quentini | | Millerozyma farinosa | | | Gemella morbillorum | Haemophilus sputorum | | Naganishia albida | | | Granulicatella adiacens | Herbaspirillum huttiense | | Papilistrema laurentii | | | Granulicatella elegans | Kingella denitrificans | | Penicillium chrysogenum | | | Kocuria kristinae | Kingella kingae | | Rhodotorula mucilaginosa | | | Kocuria rhizophila | Kingella negevensis | | Saccharomyces cerevisiae | | | Kytococcus sedentarius | Kingella oralis | | Schizosaccharomyces pombe | K243013 - Page 15 of 51 {15} | On-Panel Organisms | Off-Panel Organisms | | | | | --- | --- | --- | --- | --- | | | Gram-Positive Bacteria | Gram-Negative Bacteria | Resistance Markers | Yeasts / Viruses / Parasites | | | Lactobacillus acidophilus | Legionella pneumophila | | Talaromyces marneffei | | | Lactobacillus crispatus | Leptospira interrogans | | Trichosporon asahii | | | Lactobacillus rhamnosus | Moraxella catarrhalis | | Wickerhamomyces anomalus | | | Lactococcus garvieae | Moraxella osloensis | | BK Virus | | | Lactococcus lactis | Mycobacterium tuberculosis | | Chikungunya Virus | | | Leuconostoc carnosum | Neisseria gonorrhoeae | | Cytomegalovirus | | | Leuconostoc citreum | Neisseria lactamica | | Dengue Virus | | | Leuconostoc mesenteroides | Neisseria mucosa | | Enterovirus | | | Listeria spp. | Neisseria sicca | | Epstein Barr Virus | | | Macrococcus caseolyticus | Parabacteroides distasonis | | Hepatitis A virus | | | Micrococcus luteus | Parabacteroides merdae | | Hepatitis B virus | | | Mycobacterium avium complex (MAC) | Pasteurella aerogenes | | Hepatitis C virus | | | Mycobacterium fortuitum | Pasteurella canis | | Human alphaherpesvirus 1 | | | Mycobacterium mucogenicum | Pasteurella multocida | | Human alphaherpesvirus 2 | | | Mycoplasma hominis | Pasteurella stomatis | | Human betaherpesvirus 6 | | | Mycoplasma pneumoniae | Prevotella bivia | | Human betaherpesvirus 7 | | | Nocardia farcinica | Prevotella buccae | | Human Immunodeficiency Virus | | | Parvimonas micra | Prevotella denticola | | JC Virus | | | Pediococcus acidilactici | Prevotella melaninogenica | | Measles Virus | | | Pediococcus pentosaceus | Prevotella oralis | | Mumps Virus | | | Peptostreptococcus anaerobius | Psychrobacter cryohalolentis | | Parvovirus B19 | | | Planococcus citreus | Psychrobacter immobilis | | Rubella Virus | | | Planococcus kocurii | Ralstonia mannitolilytica | | Varicella Zoster Virus | | | Propionibacterium freudenreichii | Ralstonia pickettii | | West Nile Virus | | | Propionibacterium propionicum (Arachnia propionica) | Serratia spp. | | Zika Virus | | | Rhodococcus equi | Stenotrophomonas acidaminiphila | | Plasmodium falciparum | | | Rothia dentocariosa | Stenotrophomonas nitritireducens | | Trypanosoma cruzi | | | Rothia mucilaginosa | Stenotrophomonas rhizophila | | | | | Sarcina ventriculi | Treponema pallidum | | | | | Solibacillus silvestris | Veillonella parvula | | | | | Staphylococcus spp. | Vibrio alginolyticus | | | | | Streptococcus spp. | Vibrio parahaemolyticus | | | | | Ureaplasma parvum | Vibrio vulnificus | | | | | Ureaplasma urealyticum | | | | | | Vagococcus fluvialis | | | | | | Weissella paramesenteroides | | | | K243013 - Page 16 of 51 {16} In silico analyses of potential non-specific amplification and detection predicts the following potential cross-reactivity against organisms listed in Table 6. The predicted cross-reactivity is noted in the device labeling. - Acinetobacter baumannii oligo designs are predicted to detect some strains of Acinetobacter pittii and Klebsiella pneumoniae. - Some strains of Escherichia species (E. albertii, E. coli, E. fergusonii, E. marmotae), Raoultella terrigena and Shigella species (S. boydii, S. dysenteriae, S. flexneri, S. sonnei) are predicted to produce false positive Citrobacter spp. results. - Enterobacter spp. oligo designs are predicted to detect some strains of Klebsiella huaxiensis, Leclercia adecarboxylata and Lelliottia species (Lelliottia amnigena, Lelliottia nimipressuralis). - Some strains of Aeromonas hydrophila, Aggregatibacter aphrophilus, Corynebacterium frankenforstense, Haemophilus parahaemolyticus, Haemophilus parainfluenzae and Pseudomonas wenzhouensis are predicted to produce false positive Enterobacteriaceae / Morganellaceae results. - Escherichia coli oligo designs are predicted to detect some strains of Escherichia albertii and Escherichia marmotae. - Haemophilus influenzae oligo designs are predicted to detect some strains of Haemophilus aegyptius. - Some strains of Cedecea species (C. davisae, C. lapagei, C. neteri), Enterobacter species (E. kobei, E. mori), Escherichia coli, Klebsiella species (K. aerogenes, K. electrica, K. grimontii, K. huaxiensis, K. michiganensis, K. pasteurii, K. pneumoniae, K. variicola), Kluyvera species (K. ascorbate, K. cryocrescens), Raoultella species (R. ornithinolytica, R. planticola) and unspecified Superficieibacter species are predicted to produce false positive Klebsiella oxytoca results. - Klebsiella pneumoniae oligo designs are predicted to detect some strains of Enterobacteriaceae and Klebsiella species (K. africana, K. grimontii, K. quasipneumoniae, K. quasivariicola, K. variicola). - Some strains of Cronobacter muytjensii, Klebsiella pneumoniae and Klebsiella quasipneumoniae are predicted to produce false positive Klebsiella variicola results. - Some strains of Arsenophonus species (A. apicola, A. endosymbiont, A. nasoniae), Haemophilus parahaemolyticus, Leclercia adecarboxylata, Morganella psychrotolerans, Providencia species (P. hangzhouensis, P. heimbachae, P. huaxiensis, P. manganoxydans, P. rettgeri, P. rustigianii, P. stuartii, P. vermicola), Serratia species (S. grimesii, S. proteamaculans, S. quinivorans) and Xenorhabdus species (X. bovienii, X. budapestensis, X. griffiniae, X. hominickii, X. poinarii) are predicted to produce false positive Proteus spp. results. - Pseudomonas aeruginosa oligo designs are predicted to detect some strains of Pseudomonas paraeruginosa and Pseudomonas fluorescens. - Salmonella spp. oligo designs are predicted to detect one strain of Enterobacteriaceae. - Some strains of Serratia species (S. bockelmannii, S. entomophila, S. ficaria, S. fonticola, S. grimesii, S. inhibens, S. liquefaciens, S. nematodiphila, S. nevei, S. odorifera, S. proteamaculans, S. quinivorans, S. rhizosphaerae, S. rubidaea, S. surfactantfaciens, S. symbiotica, S. ureilytica) are predicted to produce false positive Serratia marcescens results. K243013 - Page 17 of 51 {17} - *Stenotrophomonas maltophilia* oligo designs are predicted to detect some strains of *Enterobacter cloacae* and other *Stenotrophomonas* species (*S. acidaminiphila*, *S. nitritireducens*, *S. rhizophila*). - Some strains of *Serratia liquefaciens* are predicted to produce false positive *Serratia marcescens* and CTX-M results. - Some strains of *Serratia liquefaciens* and *Serratia nevei* are predicted to produce false positive *Serratia marcescens* and IMP results. - Some strains of *Serratia nevei* are predicted to produce false positive *Serratia marcescens* and OXA results. #### *Microbial Interference and Competitive Inhibition* A microbial interference and competitive inhibition study was conducted to evaluate performance of the LIAISON PLEX BCN Assay to detect on-panel organisms in the presence of highly concentrated, potentially interfering microorganisms. To evaluate competitive inhibition, a set of ten representative pairwise combinations of a lower concentration on-panel target strain (ring positive) and a second on-panel target at high concentration (ring positive + 8 hours) were evaluated to assess dual target detection. To determine if non-target organisms can interfere with detection of on-panel organisms in the same sample, a microbial interference study was conducted. The study evaluated a set of ten representative pairwise combinations of a lower concentration on-panel target strains (ring positive) mixed with seven potentially interfering off-panel organisms at high concentration (~1.0E+08 CFU/mL). The on-panel pairwise strain mixtures were chosen to be representative of clinically relevant, polymicrobial bloodstream infections and were each tested in triplicate. Results of the microbial interference study and competitive inhibition studies are presented in **Tables 7 and 8**, respectively. Interference was not observed for the LIAISON PLEX BCN Assay when low concentration on-panel organisms are tested in the presence of high concentration interfering microbes and other on-panel organisms. **Table 7. Summary Results of Competitive Inhibition Study** | On-Panel High Concentration Target | Positivity | On-Panel Low Concentration Target | Positivity | | --- | --- | --- | --- | | *Acinetobacter baumannii* | 100% | *Citrobacter freundii* | 100% | | | 100% | *Enterobacter cloacae* | 100% | | | 100% | *Escherichia coli* | 100% | | | 100% | *Haemophilus influenzae* | 100% | | | 100% | *Klebsiella oxytoca* | 100% | | | 100% | *Klebsiella pneumoniae* | 100% | | | 100% | *Neisseria meningitidis* | 100% | | | 100% | *Proteus mirabilis* | 100% | | | 100% | *Pseudomonas aeruginosa* | 100% | | *Citrobacter freundii* | 100% | *Acinetobacter baumannii* | 100% | | | 100% | *Enterobacter cloacae* | 100% | | | 100% | *Escherichia coli* | 100% | | | 100% | *Haemophilus influenzae* | 100% | | | 100% | *Klebsiella oxytoca* | 100% | | | 100% | *Klebsiella pneumoniae* | 100% | | | 100% | *Neisseria meningitidis* | 100% | | | 100% | *Proteus mirabilis* | 100% | | | 100% | *Pseudomonas aeruginosa* | 100% | | *Enterobacter cloacae* | 100% | *Acinetobacter baumannii* | 100% | | | 100% | *Citrobacter freundii* | 100% | K243013 - Page 18 of 51 {18} | On-Panel High Concentration Target | Positivity | On-Panel Low Concentration Target | Positivity | | --- | --- | --- | --- | | | 100% | Escherichia coli | 100% | | | 100% | Haemophilus influenzae | 100% | | | 100% | Klebsiella oxytoca | 100% | | | 100% | Klebsiella pneumoniae | 100% | | | 100% | Neisseria meningitidis | 100% | | | 100% | Proteus mirabilis | 100% | | | 100% | Pseudomonas aeruginosa | 100% | | Escherichia coli | 100% | Acinetobacter baumannii | 100% | | | 100% | Citrobacter freundii | 100% | | | 100% | Enterobacter cloacae | 100% | | | 100% | Haemophilus influenzae | 100% | | | 100% | Klebsiella oxytoca | 100% | | | 100% | Klebsiella pneumoniae | 100% | | | 100% | Neisseria meningitidis | 100% | | | 100% | Proteus mirabilis | 100% | | | 100% | Pseudomonas aeruginosa | 100% | | Haemophilus influenzae | 100% | Acinetobacter baumannii | 100% | | | 100% | Citrobacter freundii | 100% | | | 100% | Enterobacter cloacae | 100% | | | 100% | Escherichia coli | 100% | | | 100% | Klebsiella oxytoca | 100% | | | 100% | Klebsiella pneumoniae | 100% | | | 100% | Neisseria meningitidis | 100% | | | 100% | Proteus mirabilis | 100% | | | 100% | Pseudomonas aeruginosa | 100% | | Klebsiella oxytoca | 100% | Acinetobacter baumannii | 100% | | | 100% | Citrobacter freundii | 100% | | | 100% | Enterobacter cloacae | 100% | | | 100% | Escherichia coli | 100% | | | 100% | Haemophilus influenzae | 100% | | | 100% | Klebsiella pneumoniae | 100% | | | 100% | Neisseria meningitidis | 100% | | | 100% | Proteus mirabilis | 100% | | | 100% | Pseudomonas aeruginosa | 100% | | Klebsiella pneumoniae | 100% | Acinetobacter baumannii | 100% | | | 100% | Citrobacter freundii | 100% | | | 100% | Enterobacter cloacae | 100% | | | 100% | Escherichia coli | 100% | | | 100% | Haemophilus influenzae | 100% | | | 100% | Klebsiella oxytoca | 100% | | | 100% | Neisseria meningitidis | 100% | | | 100% | Proteus mirabilis | 100% | | | 100% | Pseudomonas aeruginosa | 100% | | Neisseria meningitidis | 100% | Acinetobacter baumannii | 100% | | | 100% | Citrobacter freundii | 100% | | | 100% | Enterobacter cloacae | 100% | | | 100% | Escherichia coli | 100% | | | 100% | Haemophilus influenzae | 100% | | | 100% | Klebsiella oxytoca | 100% | | | 100% | Klebsiella pneumoniae | 100% | | | 100% | Proteus mirabilis | 100% | | | 100% | Pseudomonas aeruginosa | 100% | | Proteus mirabilis | 100% | Acinetobacter baumannii | 100% | | | 100% | Citrobacter freundii | 100% | K243013 - Page 19 of 51 {19} | On-Panel High Concentration Target | Positivity | On-Panel Low Concentration Target | Positivity | | --- | --- | --- | --- | | | 100% | Enterobacter cloacae | 100% | | | 100% | Escherichia coli | 100% | | | 100% | Haemophilus influenzae | 100% | | | 100% | Klebsiella oxytoca | 100% | | | 100% | Klebsiella pneumoniae | 100% | | | 100% | Neisseria meningitidis | 100% | | | 100% | Pseudomonas aeruginosa | 100% | | Pseudomonas aeruginosa | 100% | Acinetobacter baumannii | 100% | | | 100% | Citrobacter freundii | 100% | | | 100% | Enterobacter cloacae | 100% | | | 100% | Escherichia coli | 100% | | | 100% | Haemophilus influenzae | 100% | | | 100% | Klebsiella oxytoca | 100% | | | 100% | Klebsiella pneumoniae | 100% | | | 100% | Neisseria meningitidis | 100% | | | 100% | Proteus mirabilis | 100% | Table 8. Summary Results of Microbial Interference Study | On-Panel Low Concentration Target | Positivity | Off-Panel High Concentration Target | Positivity | | --- | --- | --- | --- | | Acinetobacter baumannii | 100% | Bacillus cereus | 0% | | | 100% | Clostridium perfringens | 0% | | | 100% | Corynebacterium striatum | 0% | | | 100% | Cutibacterium (Propionibacterium) acnes | 0% | | | 100% | Staphylococcus aureus | 0% | | | 100% | Staphylococcus epidermidis | 0% | | | 100% | Streptococcus mitis | 0% | | Citrobacter freundii | 100% | Bacillus cereus | 0% | | | 100% | Clostridium perfringens | 0% | | | 100% | Corynebacterium striatum | 0% | | | 100% | Cutibacterium (Propionibacterium) acnes | 0% | | | 100% | Staphylococcus aureus | 0% | | | 100% | Staphylococcus epidermidis | 0% | | | 100% | Streptococcus mitis | 0% | | Enterobacter cloacae | 100% | Bacillus cereus | 0% | | | 100% | Clostridium perfringens | 0% | | | 100% | Corynebacterium striatum | 0% | | | 100% | Cutibacterium (Propionibacterium) acnes | 0% | | | 100% | Staphylococcus aureus | 0% | | | 100% | Staphylococcus epidermidis | 0% | | | 100% | Streptococcus mitis | 0% | | Escherichia coli | 100% | Bacillus cereus | 0% | | | 100% | Clostridium perfringens | 0% | | | 100% | Corynebacterium striatum | 0% | | | 100% | Cutibacterium (Propionibacterium) acnes | 0% | | | 100% | Staphylococcus aureus | 0% | | | 100% | Staphylococcus epidermidis | 0% | | | 100% | Streptococcus mitis | 0% | | Haemophilus influenzae | 100% | Bacillus cereus | 0% | | | 100% | Clostridium perfringens | 0% | | | 100% | Corynebacterium striatum | 0% | | | 100% | Cutibacterium (Propionibacterium) acnes | 0% | | | 100% | Staphylococcus aureus | 0% | | | 100% | Staphylococcus epidermidis | 0% | K243013 - Page 20 of 51 {20} | On-Panel Low Concentration Target | Positivity | Off-Panel High Concentration Target | Positivity | | --- | --- | --- | --- | | Klebsiella oxytoca | 100% | Streptococcus mitis | 0% | | | 100% | Bacillus cereus | 0% | | | 100% | Clostridium perfringens | 0% | | | 100% | Corynebacterium striatum | 0% | | | 100% | Cutibacterium (Propionibacterium) acnes | 0% | | | 100% | Staphylococcus aureus | 0% | | | 100% | Staphylococcus epidermidis | 0% | | | 100% | Streptococcus mitis | 0% | | Klebsiella pneumoniae | 100% | Bacillus cereus | 0% | | | 100% | Clostridium perfringens | 0% | | | 100% | Corynebacterium striatum | 0% | | | 100% | Cutibacterium (Propionibacterium) acnes | 0% | | | 100% | Staphylococcus aureus | 0% | | | 100% | Staphylococcus epidermidis | 0% | | | 100% | Streptococcus mitis | 0% | | Neisseria meningitidis | 100% | Bacillus cereus | 0% | | | 100% | Clostridium perfringens | 0% | | | 100% | Corynebacterium striatum | 0% | | | 100% | Cutibacterium (Propionibacterium) acnes | 0% | | | 100% | Staphylococcus aureus | 0% | | | 100% | Staphylococcus epidermidis | 0% | | | 100% | Streptococcus mitis | 0% | | Proteus mirabilis | 100% | Bacillus cereus | 0% | | | 100% | Clostridium perfringens | 0% | | | 100% | Corynebacterium striatum | 0% | | | 100% | Cutibacterium (Propionibacterium) acnes | 0% | | | 100% | Staphylococcus aureus | 0% | | | 100% | Staphylococcus epidermidis | 0% | | | 100% | Streptococcus mitis | 0% | | Pseudomonas aeruginosa | 100% | Bacillus cereus | 0% | | | 100% | Clostridium perfringens | 0% | | | 100% | Corynebacterium striatum | 0% | | | 100% | Cutibacterium (Propionibacterium) acnes | 0% | | | 100% | Staphylococcus aureus | 0% | | | 100% | Staphylococcus epidermidis | 0% | | | 100% | Streptococcus mitis | 0% | ### Endogenous/Exogenous Interference An interfering substances study was conducted to assess the performance of the LIAISON PLEX BCN Assay in the presence of potentially interfering substances that may be present in blood culture specimens. A set of four representative on-panel target organisms were tested in the presence of six interfering substances across five replicates. A panel containing negative blood matrix was also tested to assess the risk of false positive results in the presence of potentially interfering substances. No interference was observed (Table 9). Table 9. Summary Results of Interfering Substances Study | Interfering Substance | Concentration | Target | Positivity | | --- | --- | --- | --- | | Unconjugated Bilirubin | 20 mg/dL | Acinetobacter baumannii | 100% | | | | Escherichia coli | 100% | | | | Haemophilus influenzae | 100% | | | | Neisseria meningitidis | 100% | | | | Negative Control | 0% | K243013 - Page 21 of 51 {21} | Conjugated Bilirubin | 20 mg/dL | *Acinetobacter baumannii* | 100% | | --- | --- | --- | --- | | | | *Escherichia coli* | 100% | | | | *Haemophilus influenzae* | 100% | | | | *Neisseria meningitidis* | 100% | | | | Negative Control | 0% | | Hemoglobin | 14 g/L | *Acinetobacter baumannii* | 100% | | | | *Escherichia coli* | 100% | | | | *Haemophilus influenzae* | 100% | | | | *Neisseria meningitidis* | 100% | | | | Negative Control | 0% | | Intralipid | 3000 mg/dL | *Acinetobacter baumannii* | 100% | | | | *Escherichia coli* | 100% | | | | *Haemophilus influenzae* | 100% | | | | *Neisseria meningitidis* | 100% | | | | Negative Control | 0% | | γ-globulin | 6 g/dL | *Acinetobacter baumannii* | 100% | | | | *Escherichia coli* | 100% | | | | *Haemophilus influenzae* | 100% | | | | *Neisseria meningitidis* | 100% | | | | Negative Control | 0% | | Sodium polyanetholsulfonate (SPS) | 0.25% w/v | *Acinetobacter baumannii* | 100% | | | | *Escherichia coli* | 100% | | | | *Haemophilus influenzae* | 100% | | | | *Neisseria meningitidis* | 100% | | | | Negative Control | 0% | # 4. Assay Reportable Range: This study is not applicable as the test device is a qualitative assay. # 5. Traceability, Stability and Expected Values (Controls, Calibrators, or Methods): # *Quality Controls* Each BCN Assay Cartridge includes internal controls to ensure performance of sample preparation and detection. The internal extraction control is present in the lysis tube when the sample is added. Sample preparation is initiated, and the extraction control assesses extraction, nucleic acid recovery, and detection. A post-extraction hybridization control serves as an indicator of successful hybridization. Internal control results are reported as Pass or Fail on the printed reports. Internal controls must generate a signal above threshold in each internal reaction for the system to report a valid test result. Positive and negative external controls are not provided but are recommended to be tested with each new lot or shipment of reagents, or monthly (whichever occurs first). Negative blood matrix can be used as the negative control. Previously characterized positive samples or verified negative blood matrix spiked with well characterized organisms may be used as the external positive control. External controls should be used in accordance with laboratory protocols and in accordance with local, state, and federal accrediting organizations, as applicable. # *Sample Stability* A sample stability study was performed to establish the recommended storage conditions for positive blood cultures prior to testing with the LIAISON PLEX Gram-Negative Blood Culture (BCN) Assay. A representative panel of four on-panel organisms as well as a negative blood matrix control were tested. Fresh samples (ring positive and ring positive + 8 K243013 - Page 22 of 51 {22} hours) were stored at various conditions, refrigerated (2° to 8°C) and room temperature (15°C to 30°C), and tested over various timepoints. Results of the sample stability study are presented in **Tables 10 and 11** below. **Table 10. Summary Results of Sample Stability (Refrigerated, 2° to 8°C)** | Organism/Resistance Marker | Condition | % Positivity At Timepoints | | | | | --- | --- | --- | --- | --- | --- | | | | T0 | T1 (24 hours) | T2 (72 hours) | T3 (7 days) | | *Haemophilus influenzae* | Ring Positive | 100% (20/20) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | | Ring Positive +8hr | 100% (20/20) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | *Acinetobacter baumannii* + OXA | Ring Positive | 100% (20/20) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | | Ring Positive +8hr | 100% (20/20) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | *Escherichia coli* + MCR | Ring Positive | 100% (20/20) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | | Ring Positive +8hr | 100% (20/20) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | *Neisseria meningitidis* | Ring Positive | 100% (20/20) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | | Ring Positive +8hr | 100% (20/20) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | Negative Blood Matrix | NA | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/3) | **Table 11. Summary Results of Sample Stability (RT, 15°C to 30°C)** | Organism/Resistance Marker | Condition | % Positivity At Timepoints | | | | | --- | --- | --- | --- | --- | --- | | | | T0 | T1 (24 hours) | T2 (72 hours) | T3 (7 days) | | *Haemophilus influenzae* | Ring Positive | 100% (20/20) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | | Ring Positive +8hr | 100% (20/20) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | *Acinetobacter baumannii* + OXA | Ring Positive | 100% (20/20) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | | Ring Positive +8hr | 100% (20/20) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | *Escherichia coli* + MCR | Ring Positive | 100% (20/20) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | | Ring Positive +8hr | 100% (20/20) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | *Neisseria meningitidis* | Ring Positive | 100% (20/20) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | | Ring Positive +8hr | 100% (20/20) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | Negative Blood Matrix | NA | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/3) | #### *Fresh Versus Frozen Study* The performance of the LIAISON PLEX BCN Assay with fresh and frozen samples was evaluated by testing four on-panel organisms (ring positive and ring positive + 8 hours) as K243013 - Page 23 of 51 {23} well as a negative blood matrix control under five conditions: Initial testing of fresh samples (T0), 1 freeze-thaw cycle (T1), 2 freeze-thaw cycles (T2), 1 freeze-thaw cycle after 1 month of frozen storage (T3), and 2 freeze-thaw cycles after 1 month of frozen storage (T4). Samples were stored frozen for a minimum of 8 hours in between each freeze-thaw cycle. For the 1-month testing, material was frozen for at least 34 days prior to the first freeze-thaw. All samples demonstrated 100% agreement with the fresh sample (Table 12). **Table 12. Summary Results of Fresh Versus Frozen Study** | Representative Organisms | Condition | Initial Testing | | | 1-Month Testing | | | --- | --- | --- | --- | --- | --- | --- | | | | Fresh T(0) | 1^{st} Freeze-thaw T(1) | 2^{nd} Freeze-thaw T(2) | 1^{st} Freeze-thaw T(3) | 2^{nd} Freeze-thaw T(4) | | *Acinetobacter baumannii* | Ring Positive | 100% (12/12) | 100% (10/10) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | | Ring Positive +8hr | 100% (12/12) | 100% (10/10) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | *Escherichia coli* | Ring Positive | 100% (12/12) | 100% (10/10) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | | Ring Positive +8hr | 100% (12/12) | 100% (10/10) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | *Haemophilus influenzae* | Ring Positive | 100% (12/12) | 100% (10/10) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | | Ring Positive +8hr | 100% (12/12) | 100% (10/10) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | *Neisseria meningitidis* | Ring Positive | 100% (12/12) | 100% (10/10) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | | Ring Positive +8hr | 100% (12/12) | 100% (10/10) | 100% (10/10) | 100% (10/10) | 100% (10/10) | | Negative Blood Matrix | Ring Negative | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/3) | ## 6. Detection Limit: ### Growth and Detection Study A growth and detection study was conducted to establish the range of expected organism concentrations present in incubated blood cultures at bottle positivity (i.e., ring positive) and eight hours after ring positive. Seventeen (17) organisms representing the LIAISON PLEX Gram-Negative Blood Culture (BCN) Assay reportable targets were cultivated to ring positive and eight hours after ring positivity using a continuous monitoring blood culture system. Three unique blood bottles were tested in triplicate to assess ring positive growth and ring positive + eight hours for a total of nine bottles per organism. In addition, CFU/mL quantitation was done for each bottle. One negative blood bottle was also grown for at least five days. Results from the growth and detection study are presented in Table 13. **Table 13. Summary Results for Growth and Detection Study** | Organism | Assay Targets Reported | At Positivity | 8 Hours After Positivity | | --- | --- | --- | --- | | Mean Bottle Concentration (CFU/mL) | Agreement / Total (% Detected) | Mean Bottle Concentration (CFU/mL) | Agreement / Total (% Detected) | | *Acinetobacter baumannii* [OXA] IHMA 128307 | *Acinetobacter baumannii* | *Acinetobacter* spp. | 2.06E+08 | 9/9 (100%) | 1.63E+08 | 9/9 (100%) | K243013 - Page 24 of 51 {24} | Organism | Assay Targets Reported | At Positivity | 8 Hours After Positivity | | --- | --- | --- | --- | | Mean Bottle Concentration (CFU/mL) | Agreement / Total (% Detected) | Mean Bottle Concentration (CFU/mL) | Agreement / Total (% Detected) | | | | OXA | | | | | | *Citrobacter freundii* [VIM] IHMA 549813 | *Citrobacter* spp. | *Enterobacteriaceae* / *Morganellaceae* | VIM | 1.28E+09 | 9/9 (100%) | 1.87E+09 | 9/9 (100%) | | *Enterobacter cloacae* ATCC 35030 | *Enterobacter* spp. | *Enterobacteriaceae* / *Morganellaceae* | 1.58E+09 | 9/9 (100%) | 2.04E+09 | 9/9 (100%) | | *Escherichia coli* [MCR] NCTC 13846 | *Enterobacteriaceae* / *Morganellaceae* | *Escherichia coli* | MCR | 1.87E+09 | 9/9 (100%) | 1.65E+09 | 9/9 (100%) | | *Klebsiella oxytoca* [CTX-M] IHMA 683079 | CTX-M | *Enterobacteriaceae* / *Morganellaceae* | *Klebsiella oxytoca* | 9.69E+08 | 9/9 (100%) | 3.41E+08 | 9/9 (100%) | | *Klebsiella pneumoniae* [KPC] IHMA 629630 | *Enterobacteriaceae* / *Morganellaceae* | KPC | *Klebsiella pneumoniae* | 1.56E+09 | 9/9 (100%) | 1.58E+09 | 9/9 (100%) | | *Klebsiella variicola* ATCC BAA-830 | *Enterobacteriaceae* / *Morganellaceae* | *Klebsiella variicola* | 1.75E+09 | 9/9 (100%) | 2.06E+09 | 9/9 (100%) | | *Salmonella enteritidis* ATCC 13076 | *Enterobacteriaceae* / *Morganellaceae* | *Salmonella* spp. | 2.56E+08 | 9/9 (100%) | 1.85E+09 | 9/9 (100%) | | *Serratia marcescens* [SME] NCTC 13920 | *Enterobacteriaceae* / *Morganellaceae* | SME | *Serratia marcescens* | 2.38E+08 | 9/9 (100%) | 1.47E+09 | 12/12^{1} (100%) | | *Morganella morganii* [NDM] IHMA 605873 | *Enterobacteriaceae* / *Morganellaceae* | *Morganella morganii* | NDM | 1.68E+09 | 9/9 (100%) | 2.71E+09 | 9/9 (100%) | | *Pseudomonas aeruginosa* [IMP] IHMA 576602 | IMP | *Pseudomonas aeruginosa* | *Pseudomonas* spp. | 7.99E+08 | 9/9 (100%) | 1.26E+09 | 9/9 (100%) | | *Stenotrophomonas maltophilia* ATCC 13636 | *Stenotrophomonas maltophilia* | 4.48E+08 | 9/9 (100%) | 1.00E+09 | 9/9 (100%) | | *Proteus mirabilis* ATCC 12453 | *Enterobacteriaceae* / *Morganellaceae* | *Proteus* spp. | 2.26E+09 | 9/9 (100%) | 2.12E+09 | 9/9 (100%) | | *Haemophilus influenzae* ATCC 9007 | *Haemophilus influenzae* | 2.13E+09 | 9/9 (100%) | 1.70E+09 | 9/9 (100%) | | *Neisseria meningitidis* ATCC 43744 | *Neisseria meningitidis* | 1.88E+08 | 9/9 (100%) | 2.87E+08 | 9/9 (100%) | | Organism | Assay Targets Reported | Mean Bottle Concentration (CFU/mL) | Agreement / Total (% Detected) | | Negative Blood | None | 0.00E+00 | 0/3 (0%) | K243013 - Page 25 of 51 {25} $^{1}$ One OXA false positive was observed in initial testing. An additional set of replicates was tested resulting in 12 total replicates. ### Analytical Reactivity An analytical reactivity study was performed to evaluate the inclusivity of the LIAISON PLEX BCN Assay through testing of multiple strains representing on-panel reportable targets. All on-panel organisms (n = 246) were tested in triplicate. A summary of organism results is listed in **Table 14**. A summary of results for resistance markers is listed in **Table 15**. One hundred percent (100%) positivity was noted for all targets. **Table 14. Summary Results of Inclusivity Study for Target Organisms** | Reportable Target (Family) | Reportable Target (Genus) | Reportable Target (Species) | Organism | # Strains | % Detected | | --- | --- | --- | --- | --- | --- | | N/A | *Acinetobacter* spp. | N/A | *Acinetobacter guillouiae* | 1 | 100% | | *Acinetobacter radioresistens* | 2 | 100% | | *Acinetobacter baylyi* | 1 | 100% | | *Acinetobacter bereziniae* | 1 | 100% | | *Acinetobacter calcoaceticus* | 1 | 100% | | *Acinetobacter haemolyticus* | 1 | 100% | | *Acinetobacter indicus* | 1 | 100% | | *Acinetobacter johnsonii* | 1 | 100% | | *Acinetobacter junii* | 1 | 100% | | *Acinetobacter lwoffii* | 1 | 100% | | *Acinetobacter nosocomialis* | 1 | 100% | | *Acinetobacter pittii* | 1 | 100% | | *Acinetobacter schindleri* | 1 | 100% | | *Acinetobacter ursingii* | 1 | 100% | | *Acinetobacter* spp. | *Acinetobacter baumannii* | *Acinetobacter baumannii* | 5 | 100% | | *Enterobacteriaceae* / *Morganellaceae* | *Citrobacter* spp. | N/A | *Citrobacter amalonaticus* | 2 | 100% | | *Citrobacter braakii* | 2 | 100% | | *Citrobacter farmeri* | 1 | 100% | | *Citrobacter freundii* | 4 | 100% | | *Citrobacter gillenii* | 1 | 100% | | *Citrobacter koseri* | 5 | 100% | | *Citrobacter murliniae* | 1 | 100% | | *Citrobacter rodentium* | 1 | 100% | | *Citrobacter sedlakii* | 1 | 100% | | *Citrobacter werkmanii* | 1 | 100% | | *Citrobacter youngae* | 1 | 100% | | *Enterobacteriaceae* / *Morganellaceae* | *Enterobacter* spp. | N/A | *Enterobacter cloacae* | 10 | 100% | | *Enterobacter aerogenes* | 5 | 100% | | *Enterobacter amnigenus* | 1 | 100% | | *Enterobacter asburiae* | 1 | 100% | | *Enterobacter bugandensis* | 1 | 100% | | *Enterobacter cancerogenus* | 1 | 100% | | *Enterobacter hormaechei* | 2 | 100% | | *Enterobacter ludwigii* | 1 | 100% | | *Enterobacteriaceae* / *Morganellaceae* | N/A | *Escherichia coli* | *Escherichia coli* | 21 | 100% | | *Shigella boydii* | 2 | 100% | | *Shigella dysenteriae* | 2 | 100% | | *Shigella flexneri* | 2 | 100% | | *Shigella sonnei* | 2 | 100% | | *Enterobacteriaceae* / *Morganellaceae* | N/A | *Klebsiella oxytoca* | *Klebsiella oxytoca* | 5 | 100% | K243013 - Page 26 of 51 {26} | Reportable Target (Family) | Reportable Target (Genus) | Reportable Target (Species) | Organism | # Strains | % Detected | | --- | --- | --- | --- | --- | --- | | *Enterobacteriaceae* / *Morganellaceae* | N/A | *Klebsiella pneumoniae* | *Klebsiella pneumoniae* | 26 | 100% | | *Enterobacteriaceae* / *Morganellaceae* | N/A | *Klebsiella variicola* | *Klebsiella variicola* | 5 | 100% | | N/A | N/A | *Haemophilus influenzae* | *Haemophilus influenzae* | 6 | 100% | | *Enterobacteriaceae* / *Morganellaceae* | N/A | *Morganella morganii* | *Morganella morganii* | 5 | 100% | | N/A | N/A | *Neisseria meningitidis* | *Neisseria meningitidis* | 10 | 100% | | *Enterobacteriaceae* / *Morganellaceae* | *Proteus* spp. | N/A | *Proteus hauseri* | 1 | 100% | | | | | *Proteus mirabilis* | 5 | 100% | | | | | *Proteus myxofaciens* | 1 | 100% | | | | | *Proteus penneri* | 1 | 100% | | | | | *Proteus vulgaris* | 2 | 100% | | N/A | *Pseudomonas* spp. | N/A | *Pseudomonas alcaligenes* | 1 | 100% | | | | | *Pseudomonas chlororaphis* | 1 | 100% | | | | | *Pseudomonas fluorescens* | 1 | 100% | | | | | *Pseudomonas luteola* | 1 | 100% | | | | | *Pseudomonas mendocina* | 1 | 100% | | | | | *Pseudomonas monteilii* | 1 | 100% | | | | | *Pseudomonas mosselii* | 1 | 100% | | | | | *Pseudomonas mucidolens* | 1 | 100% | | | | | *Pseudomonas oryzihabitans* | 1 | 100% | | | | | *Pseudomonas pseudoalcaligenes* | 1 | 100% | | | | | *Pseudomonas putida* | 1 | 100% | | | | | *Pseudomonas resinovorans* | 1 | 100% | | | | | *Pseudomonas stutzeri* | 1 | 100% | | | | | *Pseudomonas veronii* | 1 | 100% | | | | | *Pseudomonas aeruginosa* | 4 | 100% | | *Enterobacteriaceae* / *Morganellaceae* | *Salmonella* spp. | N/A | *Salmonella bongori* | 1 | 100% | | | | | *Salmonella enterica* | 12 | 100% | | | | | *Salmonella enterica* subsp. *Arizonae* | 1 | 100% | | | | | *Salmonella enterica* subsp. *diarizonae* | 1 | 100% | | | | | *Salmonella enterica* subsp. *houtenae* | 1 | 100% | | | | | *Salmonella enterica* subsp. *indica* | 1 | 100% | | N/A | N/A | *Stenotrophomonas maltophilia* | *Stenotrophomonas maltophilia* | 5 | 100% | | *Enterobacteriaceae* / *Morganellaceae* | N/A | *Serratia marcescens* | *Serratia marcescens* | 6 | 100% | | *Enterobacteriaceae* / *Morganellaceae* | N/A | N/A | *Cedecea davisae* | 2 | 100%^ | | | | | *Cedecea lapagei* | 1 | 100%^ | | | | | *Cedecea neteri* | 1 | 100% | | | | | *Cronobacter muytjensii* | 1 | 100% | | | | | *Cronobacter sakazakii* | 2 | 100% | | | | | Enteric group 137 | 1 | 100% | | | | | *Escherichia albertii* | 1 | 100%^ | | | | | *Escherichia fergusonii* | 1 | 100% | K243013 - Page 27 of 51 {27} | Reportable Target (Family) | Reportable Target (Genus) | Reportable Target (Species) | Organism | # Strains | % Detected | | --- | --- | --- | --- | --- | --- | | | | | *Escherichia hermanii* | 1 | 100% | | | | | *Edwardsiella tarda* | 1 | 100% | | | | | *Hafnia alvei* | 1 | 100% | | | | | *Hafnia alvei* | 1 | 100% | | | | | *Kluyvera ascorbata* | 1 | 100%^ | | | | | *Kluyvera cryocrescens* | 1 | 100% | | | | | *Kluyvera georgiana* | 1 | 100% | | | | | *Kluyvera intermedia* | 1 | 100% | | | | | *Leclercia adecarboxylata* | 1 | 100%^ | | | | | *Lelliottia nimipressuralis* | 1 | 100%^ | | | | | *Pantoea agglomerans* | 1 | 100% | | | | | *Plesiomonas shigelloides* | 1 | 100% | | | | | *Pluralibacter gergoviae* | 1 | 100% | | | | | *Providencia acalfaciens* | 1 | 100% | | | | | *Providencia heimbachae* | 1 | 100% | | | | | *Providencia rettigeri* | 1 | 100%^ | | | | | *Providencia stuartii* | 1 | 100%^ | | | | | *Rahnella aquatillis* | 1 | 100% | | | | | *Raoultella ornithinolytica* | 1 | 100%^ | | | | | *Raoultella planticola* | 1 | 100%^ | | | | | *Raoultella terrigena* | 1 | 100%^ | | | | | *Serratia plymuthica* | 1 | 100% | | | | | *Serratia grimesii* | 1 | 100%^ | | | | | *Xenorhabdus bovienii* | 1 | 100%^ | | | | | *Xenorhabdus poinarii* | 1 | 100% | | | | | *Yersinia enterocolitica* | 1 | 100% | | | | | *Yokenella regensburgei* | 1 | 100% | | | | | *Serratia ficaria* | 1 | 100%^ | | | | | *Serratia fonticola* | 1 | 100%^ | | | | | *Serratia liquefaciens* | 1 | 100%^ | | | | | *Serratia proteamaculans* | 1 | 0%^ | | | | | *Serratia rubidaea* | 1 | 100%^ | ^ Tested strain gave expected target call and additional cross-reactive target call. See Table 16. **Table 15. Summary Results of Inclusivity Study for Target Resistance Markers** | Reportable Target (Resistance Marker) | Organism | # Strains | % Detected | | --- | --- | --- | --- | | CTX-M | *Citrobacter freundii* | 1 | 100% | | | *Enterobacter cloacae* | 5 | 100% | | | *Escherichia coli* | 14 | 100% | | | *Klebsiella oxytoca* | 2 | 100% | | | *Klebsiella pneumoniae* | 16 | 100% | | | *Kluyvera ascorbata* | 1 | 100% | | | *Kluyvera georgiana* | 1 | 100% | | | *Morganella morganii* | 1 | 100% | | | *Salmonella enterica* | 1 | 100% | | IMP | *Escherichia coli* | 1 | 100% | | | *Klebsiella pneumoniae* | 2 | 100% | | | *Proteus mirabilis* | 1 | 100% | | | *Pseudomonas aeruginosa* | 1 | 100% | | KPC | *Citrobacter freundii* | 2 | 100% | | | *Enterobacter cloacae* | 1 | 100% | | | *Enterobacter hormaechei* | 1 | 100% | | | *Klebsiella pneumoniae* | 3 | 100% | K243013 - Page 28 of 51 {28} | | *Morganella morganii* | 1 | 100% | | --- | --- | --- | --- | | MCR | *Enterobacter cloacae* | 2 | 100% | | | *Escherichia coli* | 5 | 100% | | | *Klebsiella pneumoniae* | 4 | 100% | | | *Salmonella enterica* | 2 | 100% | | NDM | *Enterobacter cloacae* | 1 | 100% | | | *Escherichia coli* | 4 | 100% | | | *Klebsiella pneumoniae* | 1 | 100% | | | *Morganella morganii* | 1 | 100% | | OXA | *Acinetobacter baumannii* | 5 | 100% | | | *Acinetobacter radioresistens* | 2 | 100% | | | *Acinetobacter indicus* | 1 | 100% | | | *Escherichia coli* | 2 | 100% | | | *Klebsiella pneumoniae* | 5 | 100% | | SME | *Serratia marcescens* | 6 | 100% | | VIM | *Enterobacter cloacae* | 1 | 100% | | | *Klebsiella pneumoniae* | 2 | 100% | | | *Pseudomonas aeruginosa* | 3 | 100% | Of the 246 on-panel organisms tested, 17 organisms generated additional target calls that align with known cross-reactivity based on *in silico* analysis, or design features. Additionally, one on-panel organism (*…
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