QIAstat-Dx BCID GN Plus AMR Panel

{0} [LOGO] FDA U.S. FOOD & DRUG ADMINISTRATION ### 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY ### I Background Information: #### A 510(k) Number K262168 #### B Applicant QIAGEN GmbH #### C Proprietary and Established Names QIAstat-Dx BCID GN Plus AMR Panel #### 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 | ### II Submission/Device Overview: #### A Purpose for Submission: To obtain substantial equivalence determination for the QIAstat-Dx BCID GN Plus AMR Panel #### B Measurand: Nucleic acid sequences from the following gram-negative bacteria and fungi, and antimicrobial resistance markers: - Acinetobacter calcoaceticus-baumannii complex - Bacteroides fragilis - Enterobacter spp. - Escherichia coli - Haemophilus influenzae - Klebsiella spp. - Neisseria meningitidis (encapsulated) Food and Drug Administration 10903 New Hampshire Avenue Silver Spring, MD 20993-0002 {1} - Proteus spp. - Pseudomonas aeruginosa - Salmonella spp. - Serratia spp. - Stenotrophomonas maltophilia - Pan Enterobacterales - aac(6')-lb - ampC (CMY/LAT/DHA/FOX) - armA - CTX-M - GES - IMP - KPC - NDM - OXA-23 - OXA-24/40 - OXA-48-like - OXA-58 - PER - SHV - SPM - TEM - VEB - VIM ### C Type of Test: A multiplexed nucleic acid test intended for use with the QIAstst-Dx Analyzer 2.0 for the qualitative in vitro detection and identification of nucleic acids from gram-negative bacteria and fungi in a positive blood culture media sample ### III Intended Use/Indications for Use: #### A Intended Use(s): See Indications for Use below. #### B Indication(s) for Use: The QIAstat-Dx BCID GN Plus AMR Panel is an automated qualitative multiplexed real-time PCR-based in vitro diagnostic test intended for use with the QIAstat-Dx Analyzer 2.0. The QIAstat-Dx BCID GN Plus AMR Panel is capable of simultaneous detection and identification of multiple gram-negative bacterial targets and selected genetic determinants associated with antimicrobial resistance. The following gram-negative bacteria are identified and differentiated using the QIAstat-Dx BCID GN Plus AMR Panel: - Acinetobacter calcoaceticus-baumannii complex - Bacteroides fragilis K262168 - Page 2 of 94 {2} - Enterobacter spp. - Escherichia coli - Haemophilus influenzae - Klebsiella spp. - Neisseria meningitidis (encapsulated) - Proteus spp. - Pseudomonas aeruginosa - Salmonella spp. - Serratia spp. - Stenotrophomonas maltophilia - Pan Enterobacterales The QIAstat-Dx BCID GN Plus AMR Panel contains assays for the detection of the following genetic determinants of resistance: - aac(6')-lb - ampC (CMY/LAT/DHA/FOX) - armA - CTX-M - GES - IMP - KPC - NDM - OXA-23 - OXA-24/40 - OXA-48-like - OXA-58 - PER - SHV - SPM - TEM - VEB - VIM The QIAstat-Dx BCID GN Plus AMR Panel contains assays for the detection of genetic determinants associated with resistance to aminoglycosides (aac(6')-lb and armA), penicillins and cephalosporins (ampC [CMY/LAT/DHA/FOX], CTX-M, GES, PER, SHV, TEM, and VEB), and carbapenems (GES, IMP, KPC, NDM, OXA-23, OXA-24/40, OXA-48-like, OXA-58, SPM, and VIM). The antimicrobial resistance gene detected may or may not be associated with the agent responsible for disease. Negative results for certain antimicrobial resistance assays do not indicate susceptibility, as multiple mechanisms of resistance exist. The QIAstat-Dx BCID GN Plus AMR Panel is intended for use in patients suspected of having bacterial infections identified as gram-negative via Gram stain in positive blood culture samples. The test is performed on blood culture samples identified as positive by a continuous monitoring blood culture system that demonstrates the presence of gram-negative bacteria via Gram stain. The QIAstat-Dx BCID GN Plus AMR Panel is intended to aid in the diagnosis of specific pathogens K262168 - Page 3 of 94 {3} of bloodstream infection and select determinants of antimicrobial-resistance. Results should be used in conjunction with additional laboratory testing, e.g. sub-culturing of positive blood cultures for identification of organisms not detected by QIAstat-Dx BCID GN Plus AMR Panel and susceptibility testing and clinical presentation must be taken into consideration for patient management decisions. Negative results do not preclude the presence of pathogens that are not detected by this test. Positive QIAstat-Dx BCID GN Plus AMR Panel results do not rule out infection or co-infection with organisms not included in the QIAstat-Dx BCID GN Plus AMR Panel. Negative AMR results do not preclude the presence of other antimicrobial resistance mechanisms, and the absence of a resistance gene does not necessarily predict susceptibility to a particular drug nor predict therapeutic failure. # **C Special Conditions for Use Statement(s):** Rx - For Prescription Use Only For *in vitro* Diagnostic Use Only # **D Special Instrument Requirements:** QIAstat-Dx Analyzer 2.0 # **IV Device/System Characteristics:** # **A Device Description:** The QIAstat-Dx BCID GN Plus AMR Panel is an automated, qualitative, multiplex real-time PCR-based *in vitro* diagnostic (IVD) test used with the QIAstat-Dx Analyzer 2.0. It is designed for the simultaneous detection and identification of 13 gram-negative bacterial targets and 18 genetic determinants of antimicrobial resistance (AMR) directly from positive blood culture samples confirmed gram-negative by Gram stain. The QIAstat-Dx BCID GN Plus AMR Panel test system consist of: - The QIAstat-Dx BCID GN Plus AMR Panel cartridge kit contains six individually packed, single-use disposable cartridge, which are pre-packaged with wet and dry chemistry reagents necessary to perform sample preparation, nucleic acid amplification, and assay specific PCR detection (buffers, lyophilized master mix, air-dried primers and probes, and an internal process control using *Schizosaccharomyces pombe*) and six individually packed, single use transfer pipettes. - QIAstat-Dx Analyzer 2.0, consisting of one Operational Module PRO and up to four Analytical Modules with software v1.6 or higher. The QIAstat-Dx Application Software is embedded in the QIAstat-Dx Analyzer 2.0. *Calibrators and Controls:* Negative and positive external controls are recommended but not provided with the QIAstat-Dx BCID GN Plus AMR Panel. An internal control ("IC") within the cartridge which provides a full process control covering lysis, nucleic acid purification, and DNA amplification. The IC is *Schizosaccharomyces pombe* and is in the IC cavity and mixed with the sample during sample preparation (See "**Principle of Operation**"). The QIAstat-Dx Analyzer 2.0 is provided factory calibrated and does not require user calibration. The QIAstat- K262168 - Page 4 of 94 {4} Dx Analyzer 2.0 includes self-check controls to verify the performance of all sensors and actuators and alert the user in case of failure # **B Principle of Operation:** The QIAstat-Dx BCID GN Plus AMR Panel operates as a fully automated, closed-system molecular assay designed to detect Gram-negative bacteria and antimicrobial resistance markers from positive blood culture samples. Once a blood culture sample is loaded into the disposable cartridge and inserted into the QIAstat-Dx Analyzer 2.0, the following steps occur automatically: 1. Internal Control & Proteinase K Resuspension — Following insertion of the cartridge, the IC and Proteinase K is resuspended with the buffer located in Reservoir 1. The buffer is added to the interconnected IC cavity and Proteinase K cavity and transferred repeatedly between the transfer chamber and the cavities to ensure resuspension. 2. Cell Lysis — Lysis of the sample and IC occur by a combination of mechanical (rotor + silica beads), enzymatic (Proteinase K), and chemical (guanidine-based buffers) methods. 3. Nucleic Acid Purification — DNA binds to a silica membrane, is washed to remove contaminants, and is then eluted. 4. Master Mix Rehydration — The purified eluate rehydrates the lyophilized Master Mix (containing DNA polymerase, reverse transcriptase, and dNTPs). 5. Multiplex Real-Time PCR — ~15 μL aliquots are distributed into each of 8 Reaction Chambers containing target-specific primers and probes. Within each Reaction Chamber, multiplex, real-time PCR is performed (forty PCR cycles). Increase in fluorescence (indicative of detection of each target analyte) is detected directly within each Reaction Chamber after each cycle. Results are automatically interpreted by the analyzer software and displayed as Detected / Not Detected for each target, in approximately 75 minutes. The results screen displays a message indicating that the IC “Passed” when the test is run successfully, and a message of “Failed” indicates that the internal control was not amplified. If the IC fails, detected targets are reported as “Positive with Warning” (“POSITIVE*”), and negative results are reported as “Invalid”, and testing should be repeated. # **V Substantial Equivalence Information:** # **A Predicate Device Name(s):** ePlex Blood Culture Identification Gram Negative (BCID-GN) Panel # **B Predicate 510(k) Number(s):** K182619 # **C Comparison with Predicate(s):** | Device & Predicate Device(s): | K262168 | K182619 | | --- | --- | --- | | Device Trade Name | QIAstat-Dx BCID GN Plus AMR Panel | ePlex Blood Culture Identification Gram Negative (BCID-GN) Panel | | General Device Characteristic Similarities | | | K262168 - Page 5 of 94 {5} | Intended Use/Indications For Use | The QIAstat-Dx BCID GN Plus AMR Panel is an automated qualitative multiplexed real-time PCR-based *in vitro* diagnostic test intended for use with the QIAstat-Dx Analyzer 2.0. The QIAstat-Dx BCID GN Plus AMR Panel is capable of simultaneous detection and identification of multiple gram-negative bacterial targets and selected genetic determinants associated with antimicrobial resistance. The following gram-negative bacteria are identified and differentiated using the QIAstat-Dx BCID GN Plus AMR Panel: - *Acinetobacter calcoaceticus-baumannii* complex - *Bacteroides fragilis* - *Enterobacter* spp. - *Escherichia coli* - *Haemophilus influenzae* - *Klebsiella* spp. - *Neisseria meningitidis* (encapsulated) - *Proteus* spp. - *Pseudomonas aeruginosa* - *Salmonella* spp. - *Serratia* spp. - *Stenotrophomonas maltophilia* - Pan Enterobacterales The QIAstat-Dx BCID GN Plus AMR Panel contains assays for the detection of the following genetic determinants of resistance: - *aac(6')-lb* - *ampC* (CMY/LAT/DHA/FOX) - *armA* - CTX-M - GES - IMP - KPC - NDM | The GenMark ePlex Blood Culture Identification Gram-Negative (BCID-GN) Panel is a qualitative nucleic acid multiplex in vitro diagnostic test intended for use on GenMark's ePlex Instrument for simultaneous qualitative detection and identification of multiple potentially pathogenic gram-negative bacterial organisms and select determinants associated with antimicrobial resistance in positive blood culture. In addition, the ePlex BCID-GN Panel is capable of detecting several gram-positive bacteria (Pan Gram-Positive assay) and several Candida species (Pan Candida assay). The ePlex BCID-GN Panel is performed directly on blood culture samples identified as positive by a continuous monitoring blood culture system and which contain gram-negative organism. The following bacterial organisms and genes associated with antibiotic resistance are identified using the ePlex BCID-GN Panel: *Acinetobacter baumannii*, *Bacteroides fragilis*, *Citrobacter*, *Cronobacter sakazakii*, *Enterobacter cloacae* complex, *Enterobacter* (non-cloacae complex), *Escherichia coli*, *Fusobacterium necrophorum*, *Fusobacterium nucleatum*, *Haemophilus influenzae*, *Klebsiella oxytoca*, *Klebsiella pneumoniae* group, *Morganella morganii*, *Neisseria meningitidis*, *Proteus*, *Proteus mirabilis*, *Pseudomonas aeruginosa*, *Salmonella*, *Serratia*, *Serratia marcescens*, *Stenotrophomonas maltophilia*, CTX-M (blaCTX-M), IMP (blaIMP), KPC (blaKPC), NDM (blaNDM), OXA (blaOXA) (OXA-23 and OXA-48 groups only), and VIM (blaVIM). The ePlex BCID-GN Panel contains assays for the detection of genetic determinants associated with resistance to antimicrobial agents including CTX-M(blaCTX-M), which is associated with resistance to extended spectrum beta-lactamase (ESBL)- | | --- | --- | --- | K262168 - Page 6 of 94 {6} | | - OXA-23 - OXA-24/40 - OXA-48-like - OXA-58 - PER - SHV - SPM - TEM - VEB - VIMThe QIAstat-Dx BCID GN Plus AMR Panel contains assays for the detection of genetic determinants associated with resistance to aminoglycosides (aac(6')-Ib and armA), penicillins and cephalosporins (ampC [CMY/LAT/DHA/FOX], CTX-M, GES, PER, SHV, TEM, and VEB), and carbapenems (GES, IMP, KPC, NDM, OXA-23, OXA-24/40, OXA-48-like, OXA-58, SPM, and VIM). The antimicrobial resistance gene detected may or may not be associated with the agent responsible for disease. Negative results for certain antimicrobial resistance assays do not indicate susceptibility, as multiple mechanisms of resistance exist. The QIAstat-Dx BCID GN Plus AMR Panel is intended for use in patients suspected of having bacterial infections identified as gram-negative via Gram stain in positive blood culture samples. The test is performed on blood culture samples identified as positive by a continuous monitoring blood culture system that demonstrates the presence of gram-negative bacteria via Gram stain. The QIAstat-Dx BCID GN Plus AMR Panel is intended to aid in the diagnosis of specific pathogens of bloodstream infection and select determinants of antimicrobial-resistance. Results should be used in conjunction with additional | mediated resistance to penicillins, cephalosporins and monobactams, as well as OXA (blaOXA) (OXA-23 and OXA-48 groups only), KPC (blaKPC), and metallo-beta-lactamases IMP (blaIMP), VIM (blaVIM), and NDM (blaNDM), which is associated with carbapenemase-mediated resistance. The antimicrobial resistance gene detected may or may not be associated with the agent responsible for disease. Negative results for these select antimicrobial resistance assays do not indicate susceptibility, as there are multiple mechanisms of resistance in gram-negative bacteria. The ePlex BCID-GN Panel also contains targets designed to detect a broad range of organisms with a potentially misleading Gram stain result or organisms that may be missed by Gram staining altogether, for example in the case of co-infections. These include a broad Pan Gram-Positive assay (which is designed to detect Bacillus cereus group, Bacillus subtilis group, Enterococcus, Staphylococcus, and Streptococcus), as well as a Pan Candida assay, which is designed to detect four Candida species: Candida albicans, Candida glabrata, Candida krusei, and Candida parapsilosis. The detection and identification of specific bacterial and fungal nucleic acids from individuals exhibiting signs and/or symptoms of bloodstream infection aids in the diagnosis of bloodstream infection when used in conjunction with other clinical information. The results from the ePlex BCID-GN Panel are intended to be interpreted in conjunction with Gram stain results and should not be used as the sole basis for diagnosis, treatment, or other patient management decisions. Negative results in the setting of a suspected bloodstream infection may be due | | --- | --- | --- | K262168 - Page 7 of 94 {7} | | laboratory testing, e.g. sub-culturing of positive blood cultures for identification of organisms not detected by QIAstat-Dx BCID GN Plus AMR Panel and susceptibility testing and clinical presentation must be taken into consideration for patient management decisions. Negative results do not preclude the presence of pathogens that are not detected by this test. Positive QIAstat-Dx BCID GN Plus AMR Panel results do not rule out infection or co-infection with organisms not included in the QIAstat-Dx BCID GN Plus AMR Panel. Negative AMR results do not preclude the presence of other antimicrobial resistance mechanisms, and the absence of a resistance gene does not necessarily predict susceptibility to a particular drug nor predict therapeutic failure. | to infection with pathogens that are not detected by this test. Positive results do not rule out co-infection with other organisms; the organism(s) detected by the ePlex BCID-GN Panel may not be the definite cause of disease. Additional laboratory testing (e.g. sub-culturing of positive blood cultures for identification of organisms not detected by ePlex BCID-GN Panel and for susceptibility testing, differentiation of mixed growth, and association of antimicrobial resistance marker genes to a specific organism) and clinical presentation must be taken into consideration in the final diagnosis of bloodstream infection. | | --- | --- | --- | | Specimen Type | Same | Blood culture samples identified as positive by a continuous monitoring blood culture system | | Blood Culture Bottles | Same | BD BACTEC Plus Aerobic BD BACTEC Plus Anaerobic BD BACTEC Standard Aerobic BD BACTEC Standard Anaerobic BD BACTEC Peds Plus BD BACTEC Lytic Anaerobic bioMérieux BACT/ALERT SA Standard Aerobic bioMérieux BACT/ALERT SN Standard Anaerobic bioMérieux BACT/ALERT FA Plus bioMérieux BACT/ALERT FN Plus bioMérieux BACT/ALERT PF Plus Thermo Scientific VersaTREK REDOX 1 EZ Draw Aerobic Thermo Scientific VersaTREK REDOX 2 EZ Draw Anaerobic | K262168 - Page 8 of 94 {8} | Analyte Detected | Same | DNA | | --- | --- | --- | | Technological Principles | Same | Multiplex nucleic acid amplification test (NAAT) | | General Device Characteristic Differences | | | | Pathogens Detected | Gram-negative Bacteria:Acinetobacter calcoaceticus-baumannii complexBacteroides fragilisEnterobacter spp.Escherichia coliHaemophilus influenzaeKlebsiella spp.Neisseria meningitidis (encapsulated)Proteus spp.Pseudomonas aeruginosaSalmonella spp.Serratia spp.Stenotrophomonas maltophiliaPan Enterobacterales | Gram-negative Bacteria:Acinetobacter baumanniiBacteroides fragilisCitrobacterCronobacter sakazakiiEnterobacter cloacae complex,Enterobacter (non-cloacae complex)Escherichia coliFusobacterium necrophorumFusobacterium nucleatumHaemophilus influenzaeKlebsiella oxytocaKlebsiella pneumoniae groupMorganella morganiiNeisseria meningitidis (encapsulated)Proteus mirabilisProteus spp.Pseudomonas aeruginosaSalmonella spp.Serratia marcescensSerratia spp.Stenotrophomonas maltophiliaGram-positive Bacteria:Pan Gram-Positive | | Antimicrobial Resistance Genes Detected | aac(6')-lbampC (CMY/LAT/DHA/FOX)armACTX-MGESIMPKPCNDMOXA-23OXA-24/40OXA-48-like | CTX-MIMPKPCNDMOXA-23OXA-48-likeVIM | K262168 - Page 9 of 94 {9} | | OXA-58 PER SHV SPM TEM VEB VIM | | | --- | --- | --- | | Assay Controls | One internal control is included that monitors performance of both the sample processing and PCR stages. | Each cartridge includes two types of internal controls that together monitor performance of each step of the testing process, including extraction, amplification and detection processes. | | Instrumentation | QIAstat-Dx Analyzer 2.0 | GenMark ePlex Instrument | | Software Interface Result Reporting | • QIAstat-Dx Analyzer's Software • The QIAstat-Dx BCID GN Plus AMR Panel Assay Definition File ("ADF") | • GenMark ePlex System Software • GenMark ePlex BCID-GN Panel Software | ## VI Standards/Guidance Documents Referenced: ISO Standards: - ISO 14971, Third Edition (2019-12) — Medical devices – Application of risk management to medical devices. Listed as General Use (Recognition No. 5-125). This is the foundational risk management standard applied across the submission. IEC Standards: - IEC 62304, Edition 1.1 (2015-06) Consolidated Version — Medical device software – Software life cycle processes. Listed as Declaration of Conformity (Recognition No. 13-79). A formal Declaration of Conformity (DoC) was issued by QIAGEN GmbH for the ADF software, covering subclauses 5.1, clause 6, and clause 8, with a Class C software safety classification. - IEC 81001-5-1, Edition 1.0 (2021-12) — Health software and health IT systems safety, effectiveness and security – Part 5-1: Security – Activities in the product life cycle. Listed as General Use (Recognition No. 13-122). - IEC 61010-1, Edition 3.1 (2017-01) Consolidated Version — Safety requirements for electrical equipment for measurement, control and laboratory use – Part 1: General requirements. Listed as General Use (Recognition No. 19-34). - IEC 60601-1-2, Edition 4.1 (2020-09) Consolidated Version — Medical electrical equipment – Part 1-2: General requirements for basic safety and essential performance – Collateral standard: Electromagnetic disturbances. Listed as General Use (Recognition No. 19-36). - IEC 62366-1, Edition 1.1 (2020-06) Consolidated Version — Medical devices – Part 1: Application of usability engineering to medical devices. Listed as General Use (Recognition No. 5-129). CLSI Guidelines (Referenced in Analytical Testing): K262168 - Page 10 of 94 {10} - CLSI EP17-A2 — Referenced in the context of the linearity justification; defines linearity as the ability to provide measured quantity values proportional to the measurand (cited to explain inapplicability to this qualitative assay). - CLSI EP06-A — Referenced to justify that reportable range does not apply to qualitative clinical assays. Clinical Practice Standards (Referenced in Statements of Compliance): - Declaration of Helsinki — Ethical principles governing clinical research; cited in both US and OUS Statements of Compliance. - ICH E6(R2) Good Clinical Practice — International standard for clinical trial conduct; cited in both Statements of Compliance. - EN ISO/DIS 20916:2019 — Good study practice for in vitro diagnostic medical devices; cited in both Statements of Compliance for clinical performance study conduct. ### VII Performance Characteristics (if/when applicable): #### A Analytical Performance: ##### 1. Precision/Reproducibility: A precision/reproducibility study was conducted to assess the performance of the QIAstat-Dx BCID GN Plus AMR Panel. Two positive samples (1 low "bottle positive," 1 high "bottle positive" + 24 hours) and one negative sample were prepared. Positive samples were prepared by combining representative, on-panel isolates covering six of the 13 microorganism detected by the device. Organisms were selected for gram-negative taxonomic diversity and analytical worst-case characteristics, including coverage of different bacterial families and both fermenting and non-fermenting species (Acinetobacter baumannii, Salmonella enterica, Pseudomonas aeruginosa, Proteus vulgaris, Stenotrophomonas maltophilia), as well as AMR gene targets (OXA-24/40, OXA-48-like, VEB) and Pan Enterobacterales. Stock isolates were inoculated with whole blood into blood culture bottles and incubated in a blood culture monitoring system until "positive ring" (plus 24 hours for high positive samples). The negative sample was prepared by inoculating a blood culture bottle with whole blood and an off-panel organism (Cutibacterium acnes) and incubating until "positive ring." Studies were conducted in BD BACTEC Plus Aerobic and BD BACTEC Anaerobic Lytic medium bottles. Table 1 shows the sample composition and testing strategy for the precision/reproducibility studies. Table 1: Sample Composition and Test Strategy for Reproducibility Studies | Sample | Intended Target | Organism | Source ID | | --- | --- | --- | --- | | Positive or High Positive | Acinetobacter calcoaceticus-baumannii complex OXA-24/40 | Acinetobacter baumannii | NCTC 13302 | | | Salmonella spp. OXA-48-like | Salmonella enterica | NCTC 13954 | | | Pseudomonas aeruginosa VEB | Pseudomonas aeruginosa | MRSN 12914 | | | Proteus spp. | Proteus vulgaris | ATCC 6380 | K262168 - Page 11 of 94 {11} | | Stenotrophomonas maltophilia | Stenotrophomonas maltophilia | NCTC 10499 | | --- | --- | --- | --- | | | Pan Enterobacterales | Proteus vulgaris / Salmonella enterica | ATCC 6380 / NCTC 13954 | | Negative | N/A | Cutibacterium acnes | NCTC 737 | ### Site-to-Site Variability A site-to-site study was conducted at three external sites using six to seven QIAstat-Dx Analyzers per site. Runs were performed at each site over the course of six consecutive days, with two operators performing three replicates per sample each day, resulting in 36 replicates per sample per site (3 replicates × 2 operators × 6 days × 3 sites = 108 replicates at each concentration). A single lot of QIAstat-Dx BCID GN Plus AMR Panel was used for all runs in this study. A run was considered valid if no error code was detected by the software. Any run that caused an error code was repeated to obtain a valid result. Results where there was a failed internal control and at least one positive target in another reaction chamber ("Positive with warning") were considered invalid for analytical performance studies and were repeated to obtain valid results. Acceptance criteria were ≥95% detection for positive and high-positive samples and 100% negative calls for negative samples. Overall, 325 valid runs were included in the analysis. For all positive samples tested at all three sites, 100% detection was obtained for all expected targets (108/108 detected positive hits) (Table 2). No false positives were reported for any positive sample, and 100% negative calls were obtained for the negative samples. Table 2: Results of Site-to-Site Reproducibility Study | Organism | Target Type | Agreement with Expected Results | | | | 95% C.I. | | | --- | --- | --- | --- | --- | --- | --- | --- | | | | Site 1 | Site 2 | Site 3 | Overall | Lower | Upper | | *Acinetobacter calcoaceticus-baumannii* complex | Positive | 100% (36/36) | 100% (36/36) | 100% (36/36) | 100% (108/108) | 96.6% | 100.0% | | | High Positive | 100% (37/37) | 100% (36/36) | 100% (36/36) | 100% (109/109) | 96.7% | 100.0% | | Pan Enterobacterales | Positive | 100% (36/36) | 100% (36/36) | 100% (36/36) | 100% (108/108) | 96.6% | 100.0% | | | High Positive | 100% (37/37) (37/37) | 100% (36/36) (36/36) | 100% (36/36) (36/36) | 100% (109/109) (109/109) | 96.7% | 100.0% | | *Proteus* spp. | Positive | 100% (36/36) | 100% (36/36) | 100% (36/36) | 100% (108/108) | 96.6% | 100.0% | | | High Positive | 100% (37/37) | 100% (36/36) | 100% (36/36) | 100% (109/109) | 96.7% | 100.0% | | *Pseudomonas aeruginosa* | Positive | 100% (36/36) | 100% (36/36) | 100% (36/36) | 100% (108/108) | 96.6% | 100.0% | | | High Positive | 100% (37/37) | 100% (36/36) | 100% (36/36) | 100% (109/109) | 96.7% | 100.0% | | *Salmonella* spp. | Positive | 100% (36/36) | 100% (36/36) | 100% (36/36) | 100% (108/108) | 96.6% | 100.0% | K262168 - Page 12 of 94 {12} | | High Positive | 100% (37/37) | 100% (36/36) | 100% (36/36) | 100% (109/109) | 96.7% | 100.0% | | --- | --- | --- | --- | --- | --- | --- | --- | | Stenotrophomonas maltophilia | Positive | 100% (36/36) | 100% (36/36) | 100% (36/36) | 100% (108/108) | 96.6% | 100.0% | | | High Positive | 100% (37/37) | 100% (36/36) | 100% (36/36) | 100% (109/109) | 96.7% | 100.0% | | Resistance Markers | | | | | | | | | OXA-24/40 | Positive | 100% (36/36) | 100% (36/36) | 100% (36/36) | 100% (108/108) | 96.6% | 100.0% | | | High Positive | 100% (37/37) | 100% (36/36) | 100% (36/36) | 100% (109/109) | 96.7% | 100.0% | | OXA-48-like | Positive | 100% (36/36) | 100% (36/36) | 100% (36/36) | 100% (108/108) | 96.6% | 100.0% | | | High Positive | 100% | 100% | 100% | 100% | 96.7% | 100.0% | | VEB | Positive | 100% (36/36) | 100% (36/36) | 100% (36/36) | 100% (108/108) | 96.6% | 100.0% | | | High Positive | 100% (37/37) | 100% (36/36) | 100% (36/36) | 100% (109/109) | 96.7% | 100.0% | ### Lot-to-Lot Variability A lot-to-lot study was conducted at one site using one operator on three QIAstat-Dx Analyzers per day over the course of 16 consecutive days. Three cartridge lots were used in the study, generating a total of 90 replicates per sample per lot (270 total valid runs). Acceptance criteria were \(\geq 95\%\) detection for positive and high-positive samples and \(100\%\) negative calls for negative samples. For all positive samples, \(100\%\) detection was obtained for all expected targets (90/90 detected positive hits across all three lots) (Table 3), and \(100\%\) negative calls were observed for the negative samples. Table 3: Results of Lot-to-Lot Repeatability Study | Target | Sample | Agreement with Expected Results | | | | 95% C.I. | | | --- | --- | --- | --- | --- | --- | --- | --- | | | | Lot 1 | Lot 2 | Lot 3 | Overall | Lower | Upper | | Organisms | | | | | | | | | Acinetobacter calcoaceticus-baumannii complex | Positive | 100.0% (30/30) | 100.0% (30/30) | 100.0% (30/30) | 100.0% (90/90) | 96.0% | 100.0% | | | High Positive | 100.0% (30/30) | 100.0% (30/30) | 100.0% (30/30) | 100.0% (90/90) | 96.0% | 100.0% | | Pan Enterobacterales | Positive | 100.0% (30/30) | 100.0% (30/30) | 100.0% (30/30) | 100.0% (90/90) | 96.0% | 100.0% | | | High Positive | 100.0% (30/30) | 100.0% (30/30) | 100.0% (30/30) | 100.0% (90/90) | 96.0% | 100.0% | | Proteus spp. | Positive | 100.0% (30/30) | 100.0% (30/30) | 100.0% (30/30) | 100.0% (90/90) | 96.0% | 100.0% | | | | 100.0% | 100.0% | 100.0% | 100.0% | 96.0% | 100.0% | K262168 - Page 13 of 94 {13} | | High Positive | (30/30) | (30/30) | (30/30) | (90/90) | | | | --- | --- | --- | --- | --- | --- | --- | --- | | Pseudomonas aeruginosa | Positive | 100.0% (30/30) | 100.0% (30/30) | 100.0% (30/30) | 100.0% (90/90) | 96.0% | 100.0% | | | High Positive | 100.0% (30/30) | 100.0% (30/30) | 100.0% (30/30) | 100.0% (90/90) | 96.0% | 100.0% | | Salmonella spp. | Positive | 100.0% (30/30) | 100.0% (30/30) | 100.0% (30/30) | 100.0% (90/90) | 96.0% | 100.0% | | | High Positive | 100.0% (30/30) | 100.0% (30/30) | 100.0% (30/30) | 100.0% (90/90) | 96.0% | 100.0% | | Stenotrophomonas maltophilia | Positive | 100.0% (30/30) | 100.0% (30/30) | 100.0% (30/30) | 100.0% (90/90) | 96.0% | 100.0% | | | High Positive | 100.0% (30/30) | 100.0% (30/30) | 100.0% (30/30) | 100.0% (90/90) | 96.0% | 100.0% | | Resistance Markers | | | | | | | | | OXA-24/40 | Positive | 100.0% (30/30) | 100.0% (30/30) | 100.0% (30/30) | 100.0% (90/90) | 96.0% | 100.0% | | | High Positive | 100.0% (30/30) | 100.0% (30/30) | 100.0% (30/30) | 100.0% (90/90) | 96.0% | 100.0% | | OXA-48-like | Positive | 100.0% (30/30) | 100.0% (30/30) | 100.0% (30/30) | 100.0% (90/90) | 96.0% | 100.0% | | | High Positive | 100.0% (30/30) | 100.0% (30/30) | 100.0% (30/30) | 100.0% (90/90) | 96.0% | 100.0% | | VEB | Positive | 100.0% (30/30) | 100.0% (30/30) | 100.0% (30/30) | 100.0% (90/90) | 96.0% | 100.0% | | | High Positive | 100.0% (30/30) | 100.0% (30/30) | 100.0% (30/30) | 100.0% (90/90) | 96.0% | 100.0% | ## 2. Linearity: Not applicable, qualitative assay. ## 3. Analytical Specificity/Interference: The QIAstat-Dx BCID GN Plus AMR Panel was evaluated for analytical specificity/cross-reactivity and interference. ### Cross-Reactivity A total of 107 organisms, including those phylogenetically related to on-panel organisms and organisms likely to be present in positive blood culture specimens (e.g., skin flora, environmental contaminants), were tested to verify the absence of cross-reactivity with the candidate device (Table 4). All bacterial samples were initially prepared by spiking organisms at high concentration into Negative Blood Culture Matrix (NBCM) at a minimum concentration of ≥1.5×10⁸ CFU/mL; all fungi were tested at ≥1.50×10⁶ CFU/mL. All samples were tested in triplicate. Where cross-reactivity was observed at the initial concentration, samples were serially diluted until a concentration was reached at which no cross-reactivity was detected. Studies were conducted in BD BACTEC Plus Aerobic bottles. K262168 - Page 14 of 94 {14} Table 5 lists the organisms for which cross-reactivity was observed, the targets detected by the candidate device, and the concentration at which the organism was no longer cross-reactive on the candidate device. Table 4: List of Microorganisms Evaluated for Cross-Reactivity | Organism | Concentration Tested (CFU/mL) | Organism | Concentration Tested (CFU/mL) | | --- | --- | --- | --- | | Acinetobacter haemolyticus | 1.50E+08 | Staphylococcus epidermidis | 1.50E+08 | | Acinetobacter junii | 1.50E+08 | Streptococcus agalactiae | 1.50E+08 | | Acinetobacter lwoffii | 1.50E+08 | Streptococcus anginosus | 1.50E+08 | | Aeromonas hydrophila | 1.50E+08 | Streptococcus pyogenes | 1.50E+08 | | Aeromonas salmonicida | 1.50E+08 | Acremonium kiliense | 1.74E+06 | | Aeromonas sobria | 1.50E+08 | Candida albicans | 1.53E+06 | | Eikenella corrodens | 1.50E+08 | Candida auris | 5.00E+06 | | Haemophilus haemolyticus | 1.50E+08 | Candida glabrata | 1.14E+07 | | Haemophilus parahaemolyticus | 1.50E+08 | Candida guilliermondii | 3.80E+07 | | Legionella pneumophila | 1.50E+08 | Candida kefyr | 4.72E+07 | | Neisseria flavescens | 1.50E+08 | Candida tropicalis | 1.66E+06 | | Neisseria gonorrhoeae | 1.50E+08 | Corynebacterium jeikeium | 1.92E+07^{b} | | Neisseria sicca | 1.50E+08 | Corynebacterium striatum | 1.69E+08 | | Pasteurella aerogenes | 1.50E+08 | Cryptococcus gattii | 3.10E+06 | | Pasteurella multocida subsp. multocida | 1.50E+08 | Cryptococcus neoformans | 5.30E+06 | | Pseudomonas fluorescens | 1.50E+08 | Propionibacterium/Cutibacterium acnes | 2.13E+08 | | Pseudomonas mendocina | 1.50E+08 | Fusarium solani | 3.70E+03^{b} | | Pseudomonas putida | 1.50E+08 | Mucor velutinosus | 1.00E+05^{b} | | Pseudomonas stutzeri | 1.50E+08 | Staphylococcus capitis | 7.70E+07^{b} | | Ralstonia insidiosa | 1.50E+08 | Staphylococcus lugdunensis | 9.00E+07^{b} | | Ralstonia pickettii | 1.50E+08 | Trichosporon asahii | 8.00E+06 | | Vibrio alginolyticus | 1.50E+08 | Trichosporon asteroides | 2.08E+05^{b} | | Vibrio furnissii | 1.50E+08 | Trichosporon mucoides | 4.00E+07 | | Bacteroides caccae | 1.50E+08 | Penicillium marneffei (gDNA ONLY HG3) | 2.35E+07^{a} | | Bacteroides distasonis | 1.50E+08 | Trichosporon dermatis | 5.00E+06 | | Bacteroides eggerthii | 1.50E+08 | Acinetobacter nosocomialis | 1.50E+08 | | Bacteroides merdae | 1.50E+08 | Bacteroides ureolyticus | 1.50E+08 | | Bacteroides ovatus | 1.50E+08 | Burkholderia cepacia | 1.50E+08 | | Bacteroides thetaiotaomicron | 1.50E+08 | Corynebacterium diphtheriae | 6.30E+08 | | Bacteroides vulgatus | 1.50E+08 | Lactobacillus acidophilus | 1.50E+08 | | Fusobacterium gonidiaformans | 1.50E+08 | Malassezia furfur | 5.00E+06 | | Fusobacterium necrophorum | 1.50E+08 | Rhodotorula minuta | 5.00E+06 | | Fusobacterium nucleatum | 1.50E+08 | Neisseria perflava | 1.50E+08 | | Fusobacterium varium | 1.50E+08 | Candida norvegensis | 5.00E+06 | | Bacillus cereus | 1.50E+08 | Corynebacterium urealyticum | 1.50E+08 | | Candida dublinensis | 5.00E+06 | Escherichia fergusonii^{c} | 1.50E+08 | | Candida famata | 5.00E+06 | Streptococcus pneumoniae | 1.50E+08 | | Candida krusei | 5.00E+06 | Prevotella corporis | 1.50E+08 | | Candida lusitaniae | 5.00E+06 | Prevotella intermedia | 1.50E+08 | K262168 - Page 15 of 94 {15} | Candida metapsilosis | 5.00E+06 | Prevotella nigrescens | 1.50E+08 | | --- | --- | --- | --- | | Candida orthopsilosis | 5.00E+06 | Pseudomonas alcaligenes | 1.50E+08 | | Candida parapsilosis | 5.00E+06 | Haemophilus aegyptius | 1.50E+08 | | Corynebacterium ulcerans | 1.50E+08 | Leclercia adecarboxylata^{c} | 1.50E+08 | | Enterococcus faecalis | 1.50E+08 | Rhizobium pusense | 1.50E+08 | | Enterococcus faecium | 1.50E+08 | Streptococcus galloyticus | 1.50E+08 | | Lactobacillus casei | 1.50E+08 | Aggregatibacter aphrophilus | 1.50E+08 | | Lactobacillus crispatus | 1.50E+08 | Kluyvera ascorbata^{c} | 1.50E+08 | | Lactobacillus paracasei | 1.50E+08 | Phytobacter diazotrophicus^{c} | 1.50E+08 | | Lactobacillus rhamnosus | 1.50E+08 | Phytobacter ursingii^{c} | 1.50E+08 | | Listeria innocua | 1.50E+08 | Burkholderia glumae | 1.50E+08 | | Listeria monocytogenes | 1.50E+08 | Kluyvera intermedia^{c} | 1.50E+08 | | Micrococcus luteus | 1.50E+08 | Stenotrophomonas acidaminiphila | 1.50E+08 | | Staphylococcus aureus | 1.50E+08 | Haemophilus parainfluenzae | 1.50E+08 | $^{a}$D0 concentration reported in copies/ml because these samples were derived from gDNA stocks spiked into Negative Blood Culture Matrix. $^{b}$Samples were not targeted to the intended starting test concentration of ≥1.50E+08 CFU/mL for bacteria and ≥1.50E+06 CFU/mL for fungal targets. The affected samples had a lower stock titre after growth than required for this study. All samples were derived from inoculating pre-cultured pathogen into blood culture bottles. The bottles were incubated in a blood culture instrument until 'positive ring' to obtain the 'positive' samples. $^{c}$These pathogens are part of the Enterobacterales order and a positive signal for Pan Enterobacterales is expected as confirmed by the in silico assessment as part of the "Inclusivity" Study. For six pathogens tested (Leclercia adecarboxylata EP100, Phytobacter diazotrophicus EP238, Kluyvera intermedia EP241, Kluyvera ascorbate EP237, Phytobacter ursingii EP239 and Escherichia fergusonii EP091) positive results were obtained for Pan Enterobacterales. All noted pathogens belong to the Enterobacterales order. In silico analysis as part of the "Inclusivity" Study was conducted and confirmed the assay to be inclusive of these pathogens for the Pan Enterobacterales target. Therefore, the signal cannot be considered a cross-reaction for the QIAstat-Dx BCID GN Plus AMR Panel as expected. Runs were valid and data were included in the line data. Three runs obtained for EP080, Acinetobacter nosocomialis, were excluded from the analysis because it is an on-panel pathogen and was measured in error as the scope of the work was to assess cross-reactivity using off-panel pathogens. Table 5: List of Microorganisms Where Cross-Reactivity Observed | Organism | Target Detected | Concentration (CFU/mL) | | --- | --- | --- | | Acinetobacter junii | Acinetobacter calcoaceticus-baumannii complex | 3.27E+04 | | Leclercia adecarboxylata | Enterobacter spp. | 3.16E+05 | | Pseudomonas mendocina | Pseudomonas aeruginosa | 2.88E+05 | | Stenotrophomonas acidaminiphila | Stenotrophomonas maltophilia | 7.50E+05 | NOTE: Cross-reactivity was confirmed by in vitro testing at the initial concentration and serial dilution. Concentration shown is the lowest tested concentration at which all three replicates were negative (no cross-reactivity). Three additional organisms (Kluyvera ascorbata / CTX-M; Escherichia fergusonii / TEM; Haemophilus aegyptius / H. influenzae) produced unexpected signals but cross-reactivity was discarded upon investigation (on-panel gene carriers or expected subgroup targets). ### In silico Analysis Primers and probes included in the QIAstat-Dx BCID GN Plus AMR Panel were analyzed to evaluate their specificity using BLAST homology analysis run against the Core nucleotide database from The National Center for Biotechnology Information (NCBI) database. A dedicated analysis was performed to determine the specificity of the PCR assays detecting AMR genes. Results of the in silico analysis are shown in Table 6 below. Target organisms for which sequences predicted potential cross-reactivity are listed. K262168 - Page 16 of 94 {16} Table 6: Summary Results of In silico Cross-Reactivity Analysis | Target | In Silico Prediction | | --- | --- | | Bacteroides fragilis | Cross-reaction with Bacteroides hominis (B. fragilis Division II now recognized as B. hominis; cross-reaction expected based on assay design). | | Enterobacter spp. | Cross-reaction with some Citrobacter species (C. braakii, C. cronae, C. europaeus, C. freundii, C. portucalensis, C. youngae), Kosakonia cowanii, Kosakonia radicincitans, and Leclercia adecarboxylata. | | Klebsiella spp. | Cross-reaction with Lelliottia amnigena (all 9 available genomes predicted to be detected; confirmed by in vitro Inclusivity Study). | | Proteus spp. | Cross-reaction with Pantoea agglomerans (52/53 complete genomes predicted to be detected). | | Escherichia coli | Cross-reaction with Escherichia ruysiae and Shigella spp. (S. boydii, S. flexneri, S. dysenteriae, S. sonnei). Rhizobium pusense predicted in silico but not confirmed in vitro. | | Stenotrophomonas maltophilia | Cross-reaction with S. muris (within S. maltophilia complex), S. acidaminiphila, S. riyadhensis, and S. tuberculopleuritidis. | | Pseudomonas aeruginosa | Cross-reaction with Pseudomonas denitrificans (in silico). P. fluorescens and P. triclosanedens discarded as cross-reactants. | | aac(6')-Ib | Cross-reaction with ANT(3")-II-AAC(6')-IId (rare fused mechanism; originally described in a single S. marcescens clinical isolate; no evidence of widespread clinical distribution). | | ampC (FOX) | Cross-reaction with MOX-9 (plasmid-mediated class C β-lactamase; identified in a single clinical/hospital instance; no human BSI reports). | | SHV | Cross-reaction with blaOKP-c-1, blaAAK-1, and LEN β-lactamase family (chromosomal β-lactamases in K. quasipneumoniae and K. variicola; confer low-level intrinsic resistance only). | | NDM | Cross-reaction with AFM β-lactamase family (rare class B MBL; BSIs involving AFM producers are exceptionally rare and limited to isolated case reports). | # Competitive Inhibition (Microbial Interference) A competitive inhibition study was conducted to evaluate the capability of the QIAstat-Dx BCID GN Plus AMR Panel to detect multiple analytes in one sample when one pathogen is present at a high concentration and another pathogen is present at a lower concentration. Twenty-one organism strains from commercial suppliers were used to create representative combinations of on-panel and off-panel organisms commonly found in polymicrobial positive blood cultures. Samples were prepared by spiking one organism at high concentration (bottle positive + 24 hours of incubation) into Negative Blood Culture Matrix (NBCM), and the second organism at a lower concentration targeted at approximately 3×LoD for that pathogen (Table 7). Studies were conducted in BD BACTEC Plus Aerobic and BD BACTEC Plus Anaerobic bottles. K262168 - Page 17 of 94 {17} Table 7: List of Tested Organism Mixtures in Competitive Inhibition Study | Mix | High Positive | | Low Positive | | | --- | --- | --- | --- | --- | | | Organism | Concentration (CFU/mL) | Organism | Concentration (CFU/mL) | | 1 | Escherichia coli (CCUG 70662) | 6.08E+08 | Klebsiella pneumoniae (CCUG 68728) | 9.00E+04 | | 1 | Klebsiella pneumoniae (CCUG 68728) | 1.40E+09 | Escherichia coli (CCUG 70662) | 1.24E+06 | | 2 | Escherichia coli (NCTC 14320) | 2.01E+09 | Proteus mirabilis (ATCC BAA-2792) | 9.00E+04 | | 2 | Proteus mirabilis (ATCC BAA-2792) | 3.17E+09 | Escherichia coli (NCTC 14320) | 1.24E+06 | | 3 | Klebsiella pneumoniae (CCUG 68728) | 1.40E+09 | Enterobacter cloacae (NCTC 14326) | 8.10E+06 | | 3 | Enterobacter cloacae (NCTC 14326) | 9.56E+08 | Klebsiella pneumoniae (CCUG 68728) | 9.00E+04 | | 4 | Escherichia coli (NCTC 14320) | 2.01E+09 | Enterobacter cloacae (NCTC 14326) | 8.10E+06 | | 4 | Enterobacter cloacae (NCTC 14326) | 9.56E+08 | Escherichia coli (NCTC 14320) | 1.24E+06 | | 5 | Pseudomonas aeruginosa (NCTC 13921) | 1.41E+09 | Proteus mirabilis (ATCC BAA-2792) | 9.00E+04 | | 5 | Proteus mirabilis (ATCC BAA-2792) | 3.17E+09 | Pseudomonas aeruginosa (NCTC 13921) | 2.52E+06 | | 6 | Acinetobacter baumannii (ATCC BAA-2801) | 1.38E+08 | Klebsiella pneumoniae (CCUG 68728) | 9.00E+04 | | 6 | Klebsiella pneumoniae (CCUG 68728) | 1.40E+09 | Acinetobacter baumannii (ATCC BAA-2801) | 5.34E+04 | | 7 | Proteus mirabilis (ATCC BAA-2792) | 3.17E+09 | Klebsiella pneumoniae (CCUG 68728) | 9.00E+04 | | 7 | Klebsiella pneumoniae (CCUG 68728) | 1.40E+09 | Proteus mirabilis (ATCC BAA-2792) | 9.00E+04 | | 8 | Acinetobacter baumannii (NCTC 13302) | 2.72E+08 | Pseudomonas aeruginosa (NCTC 13921) | 2.52E+06 | | 8 | Pseudomonas aeruginosa (NCTC 13921) | 1.41E+09 | Acinetobacter baumannii (NCTC 13302) | 5.52E+02 | | 9 | Bacteroides fragilis (CCUG 4856T) | 8.19E+09 | Escherichia coli (NCTC 14320) | 1.24E+06 | | 9 | Escherichia coli (NCTC 14320) | 2.01E+09 | Bacteroides fragilis (CCUG 4856T) | 3.90E+04 | | 10 | Pseudomonas aeruginosa (ATCC BAA-2799) | 6.62E+07 | Stenotrophomonas maltophilia (NCTC 10259) | 2.54E+08 | | 10 | Stenotrophomonas maltophilia (NCTC 10259) | 2.05E+09 | Pseudomonas aeruginosa (ATCC BAA-2799) | 8.28E+06 | | 10 | Pseudomonas aeruginosa (NCTC 13921) | 1.25E+09 | Stenotrophomonas maltophilia (NCTC 10259) | 2.54E+08 | | 11 | Klebsiella pneumoniae (CCUG 68728) | 1.40E+09 | Serratia marcescens (ATCC 14041) | 1.89E+08 | | 11 | Serratia marcescens (ATCC 14041) | 4.82E+08 | Klebsiella pneumoniae (CCUG 68728) | 9.00E+04 | | 12 | Acinetobacter baumannii (ATCC BAA-2801) | 1.38E+08 | Enterobacter cloacae (NCTC 14326) | 8.10E+06 | | 12 | Enterobacter cloacae (NCTC 14326) | 9.56E+08 | Acinetobacter baumannii (ATCC BAA-2801) | 5.34E+04 | | 13 | Cutibacterium acnes (NCTC 737) | 1.71E+08 | Escherichia coli (NCTC 14320) | 1.24E+06 | | 14 | Enterococcus faecium (ATCC BAA-2316) | 8.96E+08 | Klebsiella pneumoniae (CCUG 68728) | 9.00E+04 | | 15 | Bacillus cereus (ATCC 21769) | 1.87E+06 | Pseudomonas aeruginosa (ATCC BAA-2799) | 8.28E+06 | | 16 | Corynebacterium striatum (ATCC 43735) | 9.54E+08 | Acinetobacter baumannii (NCTC 13302) | 5.52E+02 | | 17 | Candida albicans (ATCC 10231) | 1.83E+07 | Enterobacter cloacae (NCTC 14326) | 8.10E+06 | | 18 | Micrococcus luteus (NCTC 7011) | 2.13E+08 | Serratia marcescens (ATCC 14041) | 1.89E+08 | K262168 - Page 18 of 94 {18} | 19 | Staphylococcus aureus (ATCC BAA-2313) | 2.09E+08 | Proteus mirabilis (ATCC BAA-2792) | 9.00E+04 | | --- | --- | --- | --- | --- | | 20 | Staphylococcus epidermidis (ATCC 35984) | 7.29E+08 | Bacteroides fragilis (CCUG 4856T) | 3.90E+04 | | 21 | Enterococcus faecalis (ATCC 51299) | 7.25E+08 | Stenotrophomonas maltophilia (NCTC 10259) | 2.54E+08 | ### Endogenous/Exogenous Interference An interfering substances study was conducted to evaluate whether potentially interfering endogenous or exogenous substances that may be present in positive blood culture specimens impact the performance of the QIAstat-Dx BCID GN Plus AMR Panel. A single positive sample mix was prepared by combining three (3) representative on-panel isolates Escherichia coli (NCTC 14325), Acinetobacter baumannii (CCUG 57249), and Pseudomonas aeruginosa (MRSN 12914) at approximately 3×LoD for each pathogen in Negative Blood Culture Matrix (NBCM), covering all eight Reaction Chambers of the QIAstat-Dx BCID GN Plus AMR Panel cartridge (Table 8). A negative sample was prepared by inoculating blood culture bottles with whole human blood without any pathogenic material and incubating until a negative ring was confirmed. Studies were conducted in BD BACTEC Plus Aerobic bottles. Table 8: Positive Sample Mix Composition | Pathogen | Test Concentration (CFU/mL) | | --- | --- | | Escherichia coli | 4.05E+05 | | Acinetobacter baumannii | 5.34E+04 | | Pseudomonas aeruginosa | 3.00E+07 | Interfering substances were individually spiked into the positive and negative sample mixes to achieve the test concentrations listed in Table 9 below. All targets were detected in the positive sample mix and no false positives were observed in the negative sample mix for all substances tested, with one exception: Amoxicillin at 0.075 mg/mL caused inhibition of the Acinetobacter calcoaceticus-baumannii complex target (2/3 replicates detected). Upon retesting at the lower concentration of 0.038 mg/mL, full detection (3/3) was restored with no interference observed. Table 9: List of Potentially Interfering Substances | Substance | Concentration in Blood Culture Bottle | Solvent | | --- | --- | --- | | Acetaminophen | 0.156 mg/mL | Water | | Amoxicillin | 0.075 mg/mLa | Water | | Amoxicillin trihydrate: potassium clavulanate (4:1) | 0.075 mg/mL | Water | | Amphotericin B | 0.002 mg/mL | DMSO | | Ara-C Triphosphate | 2.64 μg/mL | Water | | Biotin (Vitamin B7) | 0.00351 mg/mL | DMSO | K262168 - Page 19 of 94 {19} | Sodium hypochlorite solution (Bleach) | 5% (v/v) / 6 mg/mL | N/A | | --- | --- | --- | | Caspofungin Diacetate | 0.005 mg/mL | DMSO | | Ceftriaxone Disodium Hemiheptahydrate | 0.966 mg/mL | Water | | Ciprofloxacin | 0.01 mg/mL | DMSO | | Ethanol | 5% (v/v) / 39.4 mg/mL | N/A | | Fluconazole | 0.075 mg/mL | Ethanol | | 5-Fluorocytosine | 0.09 mg/mL | Water | | Gentamicin sulfate | 0.03 mg/mL | Water | | Heparin Sodium | 3 Units/mL | Water | | Ibuprofen | 0.219 mg/mL | DMSO | | Imipenem Monohydrate | 0.5 mg/mL | Water | | Povidone (iodinated) | 5 mg/mL | Water | | Salicylic Acid | 0.0286 mg/mL | Water | | Sodium Polyanethol Sulfonate (SPS) | 0.0025 mg/mL | Water | | Tetracycline Hydrochloride | 0.015 mg/mL | Water | | Vancomycin (hydrochloride) | 0.12 mg/mL | Water | | Bilirubin (unconjugated) | 0.25 mg/mL | Chloroform | | Cholesterol | 0.5 mg/mL | Ethanol | | D-Glucose | 5 mg/mL | Water | | γ-globulin | 60 mg/mL | Saline | | Haemoglobin | 5 mg/mL | Water | | Magnesium Sulphate (MgSO4) | 0.1 mg/mL | Water | | Human genomic DNA (hgDNA) | 10.55 μg/mL | N/A | | Human Serum Albumin | 25 mg/mL | Water | | Triglycerides (Lipid standards) | 15 mg/mL | Chloroform | | Fibrinogen | 0.5 mg/mL | Saline | #### 4. Detection Limit and Assay Reportable Range: Limit of Detection Studies Limit of detection (LoD) studies were conducted to determine the LoD for the QIAstat-Dx BCID GN Plus AMR Panel, defined as the lowest concentration of organism detected in positive blood culture samples at least 95% of the time. The LoD was determined in combined isolate mixtures and verified in single-spiked samples. Combined LoD Study Seventeen combined sample mixtures were prepared, with up to three organisms per mix, covering a minimum of two isolates for each of the 31 targets on the panel. Samples were prepared using two methods: (1) pre-cultured pathogenic material was inoculated into blood culture bottles (BD BACTEC Plus Aerobic or Anaerobic media) and incubated until K262168 - Page 20 of 94 {20} "positive ring," and (2) for seven isolates where high-titer material was insufficient, 5.0 McFarland suspensions were generated and spiked into negative blood culture matrix (NBCM). All samples were enumerated to determine CFU/mL concentration and serially diluted in NBCM to concentrations ranging from 10× to 0.01× the estimated LoD. The LoD was established at the lowest concentration yielding a detection rate of ≥95% (≥19/20 replicates), with at least two dropouts observed at the 10-fold lower concentration. For all 31 intended targets, the LoD was successfully established in at least two isolates. Table 10 summarized the LoD results. Table 10: Summary of Established LoD Equivalency | Target | Organism | LoD (CFU/mL) | Combined LoD (CFU/mL) | | --- | --- | --- | --- | | **Organisms** | | | | | *Acinetobacter calcoaceticus*-*baumannii* complex | *Acinetobacter baumannii* | 3.50E+04 | 2.31E+04 | | | *Acinetobacter baumannii* | 1.78E+04 | | | | *Acinetobacter baumannii* | 1.78E+04 | | | | *Acinetobacter baumannii* | 4.14E+04 | | | | *Acinetobacter baumannii* | 3.50E+03 | | | *Bacteroides fragilis* | *B. fragilis* | 1.30E+04 | 1.30E+04 | | | *B. fragilis* | 1.30E+04 | | | *Enterobacter* spp. | *E. asburiae* | 3.40E+05 | 1.52E+06 | | | *E. cloacae* | 2.70E+06 | | | *Escherichia coli* | *E. coli* | 3.14E+05 | 2.26E+05 | | | *E. coli* | 1.35E+05 | | | | *E. coli* | 2.30E+05 | | | *Haemophilus influenzae* | *H. influenzae* | 9.05E+04 copies/ml^{b} (9.00E+03 CFU/ml) | 5.46E+04 copies/mL^{b} | | | *H. influenzae* | 1.87E+04 copies/ml^{b} | | | *Klebsiella* spp. | *K. pneumoniae* | 3.00E+04 | 9.84E+04 | | | *K. pneumoniae* | 3.00E+04 | | | | *K. pneumoniae* | 3.00E+05 | | | | *K. pneumoniae* | 3.38E+04 | | | *Neisseria meningitidis* (encapsulated) | *Neisseria meningitidis* (encapsulated) | 1.00E+03 | 5.50E+03 | | | *Neisseria meningitidis* (encapsulated) | 1.00E+04 | | | Pan Enterobacterales | *H. paralvei* | 1.26E+07 | 6.38E+06 | | | *C. freundii* | 1.60E+05 | | | *Proteus* spp. | *P. vulgaris* | 3.00E+05 | 1.65E+05 | | | *P. mirabilis* | 3.00E+04 | | K262168 - Page 21 of 94 {21} | Pseudomonas aeruginosa | P. aeruginosa | 1.00E+07 | 2.76E+06 | | --- | --- | --- | --- | | | P. aeruginosa | 1.00E+06 | | | | P. aeruginosa | 9.60E+05 | | | | P. aeruginosa | 8.40E+05 | | | | P. aeruginosa | 1.00E+06 | | | Salmonella spp. | S. enterica\( ^{a} \) | 1.00E+04 | 7.50E+03 | | | S. enterica | 5.00E+03 | | | Serratia spp. | S. marcescens | 7.00E+06 | 3.54E+06 | | | S. liquefaciens | 7.50E+04 | | | Stenotrophomonas maltophilia | S. maltophilia | 1.63E+08 | 8.48E+07 | | | | | | | | S. maltophilia | 6.50E+06 | | | Resistance Markers | | | | | aac(6')-Ib | K. pneumoniae | 3.00E+04 | 1.94E+05 | | | P. aeruginosa | 1.00E+05 | | | | E. coli | 1.35E+04 | | | | K. pneumoniae | 3.38E+04 | | | | P. aeruginosa | 8.40E+05 | | | | P. aeruginosa | 1.00E+05 | | | | P. mirabilis | 3.00E+05 | | | | K. pneumoniae | 3.00E+04 | | | | K. pneumoniae | 3.00E+05 | | | ampC (CMY/LAT/DHA/F OX) | K. pneumoniae | 3.38E+05 | 2.66E+05 | | | P. mirabilis | 3.00E+05 | | | | C. freundii | 1.60E+05 | | | armA | Acinetobacter baumannii | 1.78E+04 | 2.58E+04 | | | K. pneumoniae | 3.38E+04 | | | CTX-M | K. pneumoniae | 3.00E+04 | 2.58E+04 | | | E. coli | 1.35E+04 | | | | K. pneumoniae | 3.38E+04 | | | GES | K. pneumoniae | 3.00E+04 | 6.50E+04 | | | P. aeruginosa | 1.00E+05 | | | IMP | E. coli | 3.14E+05 | 1.72E+05 | | | K. pneumoniae | 3.00E+04 | | | KPC | E. coli | 2.30E+05 | 2.65E+05 | | | K. pneumoniae | 3.00E+05 | | | NDM | E. coli | 1.35E+05 | 1.76E+06 | | | K. pneumoniae | 3.38E+06 | | | OXA-23 | Acinetobacter baumannii | 1.78E+04 | 2.96E+04 | | | Acinetobacter baumannii\( ^{a} \) | 4.14E+04 | | | OXA-24/40 | Acinetobacter baumannii | 1.78E+04 | 3.76E+04 | K262168 - Page 22 of 94 {22} | | Acinetobacter baumannii | 5.73E+04 | | | --- | --- | --- | --- | | OXA-48 like | E. coli | 2.30E+05 | 1.17E+05 | | | S. enterica | 5.00E+03 | | | OXA-58 | Acinetobacter baumannii | 3.50E+04 | 1.93E+04 | | | Acinetobacter baumannii | 3.50E+03 | | | PER | Acinetobacter baumannii^{a} | 4.14E+05 | 2.57E+05 | | | P. aeruginosa | 1.00E+05 | | | SHV | K. pneumoniae | 3.00E+04 | 1.66E+05 | | | K. pneumoniae | 3.00E+05 | | | | K. pneumoniae | 3.38E+04 | | | | K. pneumoniae | 3.00E+05 | | | SPM | P. aeruginosa | 9.60E+06 | 5.22E+06 | | | P. aeruginosa | 8.40E+05 | | | TEM | K. pneumoniae | 3.00E+05 | 1.75E+06 | | | E. coli | 3.14E+05 | | | | K. pneumoniae | 3.38E+06 | | | | P. mirabilis | 3.00E+06 | | | VEB | P. aeruginosa | 1.00E+07 | 5.50E+06 | | | P. aeruginosa | 1.00E+06 | | | VIM | P. aeruginosa | 1.00E+06 | 3.50E+05 | | | P. aeruginosa | 1.00E+05 | | | | E. cloacae | 2.70E+05 | | | | P. mirabilis | 3.00E+04 | | $^{a}$ An error caused additional replicate to be included in the dataset but did not have an influence on LoD determination as all (21/21) or none (0/21) of the replicates for the individual targets in this mix were positive. $^{b}$ Isolate Stock specific LoD and average LoD for H. influenzae isolates are shown in copies/ml instead of CFU/ml. LoD for H. influenzae isolate CCUG 23946 was determined to be at 9.00E+03 CFU/ml. ### Single-Spiked LoD Verification Study A separate single-spiked LoD study was conducted using 18 single-isolate samples detecting 22 intended targets to verify that LoD performance in combined multi-isolate mixes is equivalent to that in single-isolate samples. The same sample preparation methods and serial dilution scheme were used, with testing at 1× and 0.1× the expected LoD (derived from the combined LoD study). A semi-log concentration (3.16× expected LoD) was used if ≥95% detection was not achieved at 1× LoD. For 18 out of 22 intended targets, LoD equivalency between single-spiked and combined samples was confirmed. Results were compared to the Growth and Detection Study to ensure that LoDs were at or below the concentration in positive blood culture specimens. ### Growth and Detection A growth and detection study was conducted to establish the range of expected organism concentrations present in positive blood culture samples at bottle positivity ("positive ring") and to verify that the QIAstat-Dx BCID GN Plus AMR Panel can detect all panel targets in Positive Blood Culture Media. K262168 - Page 23 of 94 {23} Eighteen contrived single-spiked blood culture samples were prepared, with one representative isolate per organism, covering all pathogens and AMR genes on the panel. In some cases, additional isolates of the same pathogen were included to represent different AMR genes. Each pathogen was inoculated into three individual BD BACTEC Plus Aerobic blood culture bottles (BD BACTEC Plus Anaerobic bottles were used for Bacteroides fragilis). Bottles were incubated in a blood culture instrument until "positive ring," removed within two hours of positive detection, and tested within two hours of removal. Each bottle replicate was enumerated to determine CFU/mL concentration. Table 11 below summarizes the results from the growth and detection study. Table 11: Results for Growth and Detection Study | Target | Organism | Established LoD (CFU/mL) | Bottle Positive Concentration (CFU/mL) | | --- | --- | --- | --- | | **Organisms** | | | | | *Acinetobacter calcoaceticus-baumannii* complex | *A. baumannii* (NCTC 13305) | 3.50E+04 | 2.11E+08 | | | *A. baumannii* (ATCC BAA-2803) | 1.78E+04 | 1.65E+08 | | | *A. baumannii* (CCUG 57249) | 1.78E+04 | 6.47E+07 | | | *A. baumannii* (ATCC BAA-2801) | 4.14E+04 | 1.65E+08 | | | *A. baumannii* (ATCC BAA-2890) | 3.50E+03 | 4.03E+07 | | *Bacteroides fragilis* | *B. fragilis* (NCTC 11625) | 1.30E+04 | 5.78E+08 | | | *B. fragilis* (NCTC 8560) | 1.30E+04 | 5.78E+08 | | *Enterobacter* spp. | *E. asburiae* (ATCC 35955) | 3.40E+05 | 3.25E+08 | | | *E. cloacae* (NCTC 14326) | 2.70E+06 | 3.25E+08 | | *Escherichia coli* | *E. coli* (CCUG 67180) | 3.14E+05 | 1.36E+08 | | | *E. coli* (NCTC 14325) | 1.35E+05 | 9.23E+07 | | | *E. coli* (NCTC 14321) | 2.30E+05 | 1.36E+08 | | *Haemophilus influenzae* | *H. influenzae* (CCUG 23946) | 9.05E+04 copies/mL^{b} | 7.68E+07 | | | *H. influenzae* (CCUG 33775) | 1.87E+04 copies/mL^{b} | 7.68E+07 | | *Klebsiella* spp. | *K. pneumoniae* (ATCC BAA-3077) | 3.00E+04 | 7.70E+07 | | | *K. pneumoniae* (NCTC 14334) | 3.00E+04 | 7.70E+07 | | | *K. pneumoniae* (ATCC BAA-1705) | 3.00E+05 | 7.70E+07 | | | *K. pneumoniae* (CCUG 68728) | 3.38E+04 | 7.70E+07 | | *Neisseria meningitidis* (encapsulated) | *N. meningitidis* (ATCC 13090) | 1.00E+03 | 1.94E+08 | | | *N. meningitidis* (ATCC 13077) | 1.00E+04 | 1.94E+08 | | Pan Enterobacterales | *H. paralvei* (CCUG 59411) | 1.26E+07 | 3.25E+08 | | | *C. freundii* (CCUG 418T) | 1.60E+05 | 4.79E+07 | | *Proteus* spp. | *P. vulgaris* (ATCC 6380) | 3.00E+05 | 1.91E+08 | | | *P. mirabilis* (ATCC BAA-2792) | 3.00E+04 | 1.91E+08 | | *Pseudomonas aeruginosa* | *P. aeruginosa* (MRSN 12914) | 1.00E+07 | 2.21E+08 | | | *P. aeruginosa* (NCTC 13437) | 1.00E+06 | 3.46E+07 | | | *P. aeruginosa* (JMI 378769) | 9.60E+06 | 3.46E+07 | K262168 - Page 24 of 94 {24} | | P. aeruginosa (NCTC 13921) | 8.40E+05 | 3.46E+07 | | --- | --- | --- | --- | | | P. aeruginosa (ATCC BAA-2794) | 1.00E+06 | 2.21E+08 | | Salmonella spp. | S. enterica (ATCC 6962) | 1.00E+04 | 4.32E+08 | | | S. enterica (NCTC 13954) | 5.00E+03 | 4.32E+08 | | Serratia spp. | S. marcescens (ATCC 14041) | 7.00E+06 | 3.83E+08 | | | S. liquefaciens (ATCC 27592) | 7.50E+04 | 3.83E+08 | | Stenotrophomonas maltophilia | S. maltophilia (NCTC 10499) | 1.63E+08 | 1.57E+07c | | | S. maltophilia (NCTC 10258) | 6.50E+06 | 1.57E+07c | | ampC (CMY/LAT/DHA/FOX) | K. pneumoniae (CCUG 68728) | 3.38E+05 | 7.70E+07 | | | P. mirabilis (ATCC BAA-2792) | 3.00E+05 | 1.91E+08 | | | C. freundii (CCUG 418T) | 1.60E+05 | 4.79E+07 | | Resistance Markers | | | | | aac(6')-Ib | K. pneumoniae (ATCC BAA-3077) | 3.00E+04 | 7.70E+07 | | | P. aeruginosa (ATCC BAA-2799) | 1.00E+05 | 2.21E+08 | | | E. coli (NCTC 14325) | 1.35E+04 | 1.36E+08 | | | K. pneumoniae (CCUG 68728) | 3.38E+04 | 7.70E+07 | | | P. aeruginosa (NCTC 13921) | 8.40E+05 | 3.46E+07 | | | P. aeruginosa (ATCC BAA-2794) | 1.00E+05 | 2.21E+08 | | | P. mirabilis (ATCC BAA-2792) | 3.00E+05 | 1.91E+08 | | | K. pneumoniae (NCTC 14334) | 3.00E+04 | 7.70E+07 | | | K. pneumoniae (ATCC BAA-1705) | 3.00E+05 | 7.70E+07 | | armA | A. baumannii (ATCC BAA-2803) | 1.78E+04 | 1.65E+08 | | | K. pneumoniae (CCUG 68728) | 3.38E+04 | 7.70E+07 | | CTX-M | K. pneumoniae (ATCC BAA-3077) | 3.00E+04 | 7.70E+07 | | | E. coli (NCTC 14325) | 1.35E+04 | 9.23E+07 | | | K. pneumoniae (CCUG 68728) | 3.38E+04 | 7.70E+07 | | GES | K. pneumoniae (ATCC BAA-3077) | 3.00E+04 | 7.70E+07 | | | P. aeruginosa (ATCC BAA-2794) | 1.00E+06a | 2.21E+08 | | IMP | E. coli (CCUG 67180) | 3.14E+05 | 1.36E+08 | | | K. pneumoniae (NCTC 14334) | 3.00E+04 | 7.70E+07 | | KPC | E. coli (NCTC 14321) | 2.30E+05 | 1.36E+08 | | | K. pneumoniae (ATCC BAA-1705) | 3.00E+05 | 7.70E+07 | | NDM | E. coli (NCTC 14325) | 1.35E+05 | 9.23E+07 | | | K. pneumoniae (CCUG 68728) | 3.38E+06 | 7.70E+07 | | OXA-23 | A. baumannii (ATCC BAA-2803) | 1.78E+04 | 1.65E+08 | | | A. baumannii (ATCC BAA-2801) | 4.14E+04 | 1.65E+08 | K262168 - Page 25 of 94 {25} | OXA-24/40 | A. baumannii (CCUG 57249) | 1.78E+04 | 6.47E+07 | | --- | --- | --- | --- | | | A. baumannii (ATCC BAA-2881) | 5.73E+04 | 2.11E+08 | | OXA-48-like | E. coli (NCTC 14321) | 2.30E+05 | 1.36E+08 | | | S. enterica (NCTC 13954) | 5.00E+03 | 4.32E+08 | | OXA-58 | A. baumannii (NCTC 13305) | 3.50E+04 | 2.11E+08 | | | A. baumannii (ATCC BAA-2890) | 3.50E+03 | 4.03E+07 | | PER | A. baumannii (ATCC BAA-2801) | 4.14E+05 | 1.65E+08 | | | P. aeruginosa (ATCC BAA-2799) | 1.00E+05 | 2.21E+08 | | SHV | K. pneumoniae (ATCC BAA-3077) | 3.00E+04 | 7.70E+07 | | | K. pneumoniae (ATCC BAA-1705) | 3.00E+05 | 7.70E+07 | | | K. pneumoniae (CCUG 68728) | 3.38E+04 | 7.70E+07 | | | K. pneumoniae (NCTC 14334) | 3.00E+05 | 7.70E+07 | | SPM | P. aeruginosa (JMI 378769) | 9.60E+06 | 3.46E+07 | | | P. aeruginosa (NCTC 13921) | 8.40E+05 | 3.46E+07 | | TEM | K. pneumoniae (ATCC BAA-3077) | 3.00E+05 | 7.70E+07 | | | E. coli (CCUG 67180) | 3.14E+05 | 1.36E+08 | | | K. pneumoniae (CCUG 68728) | 3.38E+06 | 7.70E+07 | | | P. mirabilis (ATCC BAA-2792) | 3.00E+06 | 1.91E+08 | | VEB | P. aeruginosa (MRSN 12914) | 1.00E+07 | 2.21E+08 | | | P. aeruginosa (NCTC 13437) | 1.00E+06 | 3.46E+07 | | VIM | P. aeruginosa (NCTC 13437) | 1.00E+06 | 3.46E+07 | | | P. aeruginosa (ATCC BAA-2799) | 1.00E+05 | 2.21E+08 | | | E. cloacae (NCTC 14326) | 2.70E+05 | 3.25E+08 | | | P. mirabilis (ATCC BAA-2792) | 3.00E+04 | 1.91E+08 | a GES LoD: the established single-spiked LoD (1.00E+06 CFU/mL) is reported, as it is higher (less sensitive) than the combined LoD and represents the claimed IFU value. b H. influenzae LoD is expressed in copies/mL because a valid CFU/mL titre could not be determined for isolate CCUG 33775; the LoD for isolate CCUG 23946 was established at 9.00 × 10³ CFU/mL. c The LoD for S. maltophilia isolate NCTC 10499 (1.63E+08 CFU/mL) exceeds the initial mean bottle positive concentration (1.57 E+07 CFU/mL). An expanded analysis incorporating data from multiple verification studies yielded a revised mean BP concentration of 8.76E+08 CFU/mL, which exceeds the LoD for both isolates. A detection rate of 100% (9/9) was achieved for all 31 panel targets across all 18 isolates tested, meeting the ≥95% acceptance criterion. No false-positive signals for on-panel targets were observed in any replicate, demonstrating the absence of on-panel cross-reactivity in this study. ### Analytical Reactivity/Inclusivity An analytical reactivity/inclusivity study was conducted to evaluate the inclusivity of the QIAstat-Dx BCID GN Plus AMR Panel through testing of clinically relevant organism subtypes, strains, genotypes, serotypes, species, and AMR gene variants representing the genetic, temporal, and geographic diversity for each on-panel reportable target (Table 12). K262168 - Page 26 of 94 {26} Strains that did not achieve 100% detection and their corresponding test results are shown in Table 13. The study comprised both an in silico and an in vitro component, as well as an assessment of on-panel cross-reactivity. Table 12: List of Wet Tested Organism for Inclusivity | Target | Species | Catalogue No. | Strain Designation / Strain Name | Result | | --- | --- | --- | --- | --- | | Organisms | | | | | | Acinetobacter calcoaceticus-baumannii complex | A. baumannii | PA212222 | N/A | Detected | | | A. calcoaceticus | CCUG 57816 | RUH2202 | Detected | | | A. nosocomialis | CCUG 74064 | NIPH 2119 | Detected | | | A. baumannii | ATCC BAA-2801 | 1103521 | Detected | | | A. baumannii | ATCC BAA-2803 | 1125209 | Detected | | | A. baumannii | ATCC BAA-2881 | 1173844 | Detected | | | A. baumannii | ATCC BAA-2901 | 157075 | Detected | | | A. baumannii | ATCC BAA-2890 | 177579 | Detected | | | A. baumannii | ATCC BAA-2895 | N/A | Detected | | | A. baumannii | ATCC BAA-2873 | 1081925 | Detected | | | A. baumannii | CCUG 57249 | N/A | Detected | | | A. baumannii | CCUG 68164 | N/A | Detected | | | A. baumannii | NCTC 13302 | OXA-25, 327009 | Detected | | | A. baumannii | NCTC 13305 | A-15 | Detected | | | A. baumannii | NCTC 13424 | UK 20 | Detected | | | A. pittii | CCUG 72793 | EQUA 2018:03/B | Detected | | Bacteroides fragilis | B. fragilis | CCUG 4856T | EN-2, Onslow/VPI 2553 | Detected | | | B. fragilis | CCUG 25931 | LRA 048 07 85 | Detected | | | B. fragilis | NCTC 11295 | 83008 | Detected | | | B. fragilis | NCTC 11625 | N/A | Detected | | | B. fragilis | NCTC 8560 | WPMH 1 | Detected | | Enterobacter spp. | E. amnigenus (Lelliottia amnigena) | ATCC 33072 | CUETM 77-118 | Detected | | | L. nimipressuralis | CCUG 69901 | N/A | Detected | | | E. ludwigii | CCUG 26899 | N/A | Detected | | | E. ludwigii | CCUG 51323T | EN-119 | Detected | | | E. asburiae | ATCC 35955 | CDC 14-81 | Detected | | | E. cancerogenus | CCUG 48556 | strain RSTN 68352/03 | Detected | | | E. cloacae | NCTC 14326 | 29 | Detected | | | E. hormaechei subsp. xiangfangensis | CCUG 63474 | N/A | Detected | | | E. kobei | CCUG 59627 | N/A | Detected | | | E. bugandensis | CCUG 75194T | EB-247 | Detected | | | E. hormaechei subsp. oharae | CCUG 53905T | EN-314 | Detected | K262168 - Page 27 of 94 {27} | | E. hormaechei subsp. steigerwaltii | CCUG 60271 | N/A | Detected | | --- | --- | --- | --- | --- | | | E. roggenkampii | CCUG 75192T | EN-117 | Detected | | | E. hormaechei subsp. hoffmannii | NCTC 13870 | LBM 93.03.067 | Detected | | | E. cloacae | NCTC 14328 | N/A | Detected | | | E. cloacae | NCTC 14322 | 42E | Detected | | | E. cloacae subsp. dissolvens | CCUG 25230T | W-4, 8122-8 | Detected | | | E. gergoviae | CCUG 33719 | N/A | Detected | | Escherichia coli | E. coli | ATCC BAA-2774 | 1093925 | Detected | | | E. coli | ATCC BAA-2775 | 1099675 | Detected | | | E. coli | NCTC 13919 | NCTC13919, H162440763 | Detected | | | E. coli | NCTC 14320 | N/A | Detected | | | E. coli | CCUG 67180 | N/A | Detected | | | E. coli | NCTC 14321 | 7 | Detected | | | E. coli | NCTC 14325 | N/A | Detected | | | E. coli | NCTC 14333 | MLST: 167 | Detected | | | E. coli | NCTC 13462 | N/A | Detected | | | E. coli | NCTC 14339 | MLST: 361 | Detected | | | E. coli | NCTC 13351 | N/A | Detected | | | E. coli | NCTC 13352 | N/A | Detected | | Haemophilus influenzae | H. influenzae | NCTC 4560 | NCTC 4560 [18] | Detected | | | H. influenzae | CCUG 23946 | Kilian HK 394 | Detected | | | H. influenzae | CCUG 29539 | L-378 | Detected | | | H. influenzae | CCUG 33775 | 572 | Detected | | | H. influenzae | CCUG 7317 | 641 | Detected | | Klebsiella spp. | K. aerogenes | ATCC 13048 | NCDC 819-56 | Detected | | | K. pneumoniae | ATCC BAA-3072 | 979620 | Detected | | | K. pneumoniae | ATCC BAA-1705 | ART 2008133 | Detected | | | K. pneumoniae | ATCC BAA-2783 | 972863 | Detected | | | K. pneumoniae | ATCC BAA-3060 | 1150539 | Detected | | | K. pneumoniae | ATCC BAA-3063 | 1121740 | Detected | | | K. pneumoniae | ATCC BAA-3068 | 1104762 | Detected | | | K. pneumoniae | ATCC BAA-3077 | 875638 | Detected | | | K. pneumoniae | CCUG 68727 | NCTC 13442 | Detected | | | K. pneumoniae | CCUG 68728 | NCTC 13443 | Detected | | | K. pneumoniae | NCTC 14334 | 47C | Detected | | | K. pneumoniae | NCTC 14337 | 54 | Detected | | | K. pneumoniae | JMI Strain 1238756 | N/A | Detected | | | K. variicola | ATCC BAA-830 | F2R9 | Detected | | | K. oxytoca | ATCC BAA-3059 | 957532 | Detected | K262168 - Page 28 of 94 {28} | | K. pneumoniae | NCTC 13438 | N/A | Detected | | --- | --- | --- | --- | --- | | | K. pneumoniae | ATCC 700603 | K6 | Detected | | Neisseria meningitidis(Encapsulated) | N. meningitidis | ATCC 13077 | M1027 | Detected | | | N. meningitidis | ATCC 13090 | M2092 | Detected | | | N. meningitidis | CCUG 27650 | LNP 638 | Detected | | | N. meningitidis | CCUG 55208 | EQUALIS 07-040/2 | Detected | | | N. meningitidis | NCTC 8556 | 60 | Detected | | Pan Enterobacterales | Cedecea davisae | NCTC 13724 | 005, 3278-77 | Detected | | | Citrobacter braakii | ATCC 51113 | CDC 80-58 | Detected | | | Citrobacter freundii | CCUG 418T | [ATCC 13316, NCTC 9750] | Detected | | | Citrobacter koseri | ATCC 27156 | CDC 3613-63 | Detected | | | Cronobacter sakazakii | ATCC 29004 | CDC 3225-73 | Detected | | | Edwardsiella tarda | NCTC 10396 | K 349 | Detected | | | Kluyvera cryocrescens | NCTC 10483 | 2367, No.4 | Detected | | | Kosakonia quasisacchari | NCTC 14272 | GDMCC1.1570 | Detected | | | Leclercia adecarboxylata | NCTC 13032 | 1783 | Detected | | | Morganella morganii | ATCC 25829 | M4 | Detected | | | Providencia stuartii | CCUG 74678 | N/A | Detected | | | Rahnella aquatilis | CCUG 52909 | N/A | Detected | | | Raoultella terrigena | NCTC 13038 | 84, L84/CUETM 77-176 | Detected | | | Shigella sonnei | CCUG 68726 | I virulent | Detected | | | Serratia rubidaea | CCUG 35289 | N/A | Detected | | | Tatumella ptyseos | NCTC 11468 | H36 | Detected | | | Yersinia enterocolitica subsp. enterocolitica | NCTC 12982 | 33114 | Detected | | | Yokenella regensburgei | NCTC 12131 | 3349-72 | Detected | | | Citrobacter freundii | ATCC 6879 | NCTC 4143 | Detected | | | Hafnia paralvei | CCUG 59411 | SMI Oth047 | Detected | | | Citrobacter amalonaticus | CCUG 4860 | 9823 | Detected | | | Escherichia fergusonii | CCUG 18766T | CDC 0568-73 | Detected | | | Morganella morganii subsp. siboni | ATCC 51206 | CDC 3522-75 | Detected | K262168 - Page 29 of 94 {29} | | Providencia rettgeri | CCUG 14804 | 1163 | Detected | | --- | --- | --- | --- | --- | | | Raoultella ornithinolytica | NCTC 13096 | N/A | Detected | | | Shigella boydii | CCUG 49022 | P288/NCTC 12985 | Detected | | | Escherichia hermannii (Atlantibacter hermannii) | CCUG 15714T | CDC 980-72 | Not Detected | | | Kosakonia cowanii | CCUG 45998 | 888-76, KSK 246 | Not Detected | | Proteus spp. | P. mirabilis | ATCC 43071 | CDC S-17 | Detected | | | P. mirabilis | ATCC BAA-2792 | 1104843 | Detected | | | P. penneri | NCTC 12737 | 1808-73 | Detected | | | P. vulgaris | ATCC 6380 | N/A | Detected | | | P. vulgaris | NCTC 13145 | CDC PR1 | Detected | | | P. terrae/cibarius | CCUG 35385 | HSCT 3586 T1 | Detected | | | P. hauseri | CCUG 63603 | N/A | Detected | | Pseudomonas aeruginosa | P. aeruginosa | ATCC BAA-2794 | 1079232 | Detected | | | P. aeruginosa | NCTC 13437 | N/A | Detected | | | P. aeruginosa | MRSN 12914 | NR-51575 | Detected | | | P. aeruginosa | NCTC 13921 | NCTC 13921 | Detected | | | P. aeruginosa | ATCC BAA-3099 | 1079205 | Detected | | | P. aeruginosa | ATCC BAA-3108 | 853930 | Detected | | | P. aeruginosa | ATCC BAA-2796 | 1106434 | Detected | | | P. aeruginosa | JMI Strain 378769 | N/A | Detected | | | P. aeruginosa | JMI Strain 338718 | N/A | Detected | | | P. aeruginosa | JMI Strain JMI-147 | N/A | Detected | | | P. aeruginosa | JMI Strain 1227781 | N/A | Detected | | | P. aeruginosa | MRSN 6241 | NR-51550 | Detected | | | P. aeruginosa | MRSN 8141 | NR-51558 | Detected | | | P. aeruginosa | NCTC 14383 | N/A | Detected | | Salmonella spp. | S. bongori | CCUG 63587 | N/A | Detected | | | S. enterica (serovar Newport) | ATCC 6962 | NCTC 129 | Detected | | | S. enterica subsp. enterica serotype Gallinarum | ATCC 700623 | X-142 | Detected | | | S. enterica subsp. enterica serotype Seftenberg | NCTC 13953 | N/A | Detected | | | S. enterica subsp. enterica | NCTC 13954 | N/A | Detected | K262168 - Page 30 of 94 {30} | | serotype Typhimurium | | | | | --- | --- | --- | --- | --- | | | S. enterica subsp. enterica serotype Westhampton | NCTC 8549 | 1314-52 | Detected | | Serratia spp. | S. ficaria | CCUG 20935 | F.Grimont G 5779 | Detected | | | S. fonticola | CCUG 14186 | CUETM 77-165 | Detected | | | S. liquefaciens | ATCC 27592 | CDC 1284-57 | Detected | | | S. marcescens | ATCC 14041 | NRRL B-1481 | Detected | | | S. plymuthica | ATCC 7462 | [Grimont 502] | Detected | | | S. marcescens | NCTC 13382 | NRS 116-175 | Detected | | | S. odorifera | CCUG 14508 | N/A | Detected | | Stenotrophomonas maltophilia | S. maltophilia | ATCC 13637 | 810-2 | Detected | | | S. maltophilia | CCUG 72210 | N/A | Detected | | | S. maltophilia | NCTC 10258 | 560 | Detected | | | S. maltophilia | NCTC 10259 | 611 | Detected | | | S. maltophilia | NCTC 10498 | RH 661-1 | Detected | | | S. maltophilia | NCTC 10499 | RH 873-3 | Detected | | Resistane Markers | | | | | | aac(6')-Ib | P. aeruginosa | JMI Strain 378769 | aac(6')-Ib / N/A | Detected | | | P. aeruginosa | JMI Strain 338718 | aac(6')-Ib / N/A | Detected | | | P. aeruginosa | JMI Strain JMI-147 | aac(6')-Ib / N/A | Detected | | | K. pneumoniae | ATCC BAA-3068 | aac(6')-Ib3 / 1104762 | Detected | | | E. coli | ATCC BAA-2774 | aac(6')-Ib-cr / 1093925 | Detected | | | E. coli | NCTC 14320 | aac(6')-Ib / N/A | Detected | | | K. pneumoniae | ATCC BAA-3077 | aac(6')-Ib4 / 875638 | Detected | | | P. mirabilis | ATCC BAA-2792 | aac(6')-Ib7 / 1104843 | Detected | | | P. aeruginosa | ATCC BAA-2799 | aac(6')-Ib / 1145009 | Detected | | | E. coli | NCTC 13919 | aac(6')-Ib / NCTC13919 | Detected | | | A. baumannii | ATCC BAA-2901 | aac(6')-Ib3 / 157075 | Detected | | | P. aeruginosa | ATCC BAA-2794 | aac(6')-Ib / 1079232 | Detected | | | K. pneumoniae | CCUG 68728 | aac(6')-Ib / NCTC 13443 | Detected | | | P. aeruginosa | NCTC 13921 | aac(6')-Ib / NCTC 13921 | Detected | | | K. pneumoniae | NCTC 14334 | aac(6')-Ib / 47C | Detected | K262168 - Page 31 of 94 {31} | | K. pneumoniae | ATCC BAA-1705 | aac(6')-Ib / ART 2008133 | Detected | | --- | --- | --- | --- | --- | | | E. coli | NCTC 14325 | aac(6')-Ib / N/A | Detected | | | K. pneumoniae | ATCC BAA-2783 | aac(6')-Ib3, aac(6')-Ib-cr5 / 972863 | Detected | | | P. aeruginosa | ATCC BAA-3108 | aac(6')-Ib / 853930 | Detected | | | K. pneumoniae | ATCC BAA-3072 | aac(6')-Ib-cr / 979620 | Detected | | | K. pneumoniae | ATCC BAA-3060 | aac(6')-Ib3, aac(6')-Ib-cr, aac(6')-IIc / 1150539 | Detected | | | K. pneumoniae | ATCC BAA-3063 | aac(6')-Ib-cr / 1121740 | Detected | | | K. oxytoca | ATCC BAA-3059 | aac(6')-Ib / 957532 | Detected | | | E. bugandensis | CCUG 75194T | aac(6')-Ib-cr5 / EB-247 | Detected | | | P. aeruginosa | NCTC 14383 | aac(6')-Ib / N/A | Detected | | | A. baumannii | NCTC 13424 | aac(6')-Ib / UK 20 | Detected | | | P. aeruginosa | MRSN 6241 | aac(6')-Ib / N/A | Detected | | | P. aeruginosa | MRSN 8141 | aac(6')-Ib / N/A | Detected | | | K. pneumoniae | NCTC 14337 | aac(6')-Ib / 54 | Detected | | | E. coli | NCTC 13351 | aac(6')-Ib / N/A | Detected | | | K. pneumoniae | NCTC 13438 | aac(6')-Ib / N/A | Detected | | | A. baumannii | CCUG 68164 | aac(6')-Ib / N/A | Detected with reduced sensitivity | | ampC (CMY/DHA/FOX/LAT) | K. pneumoniae | ATCC BAA-3060 | ampC (DHA-7) / 1150539 | Detected | | | gBlock | ampC FOX-X77455.1 | ampC (FOX) gBlock | Detected | | | P. mirabilis | ATCC BAA-2792 | ampC (CMY-16) / 1104843 | Detected | | | Citrobacter freundii | ATCC 6879 | ampC (CMY-LAT) / NCTC 4143 | Detected | | | Citrobacter braakii | ATCC 51113 | ampC (CMY-2) / CDC 80-58 | Detected | | | S. enterica subsp. enterica serotype Seftenberg | NCTC 13953 | ampC (CMY-LAT) | Detected | | | K. pneumoniae | CCUG 68728 | ampC (CMY-LAT), ampC (DHA) / NCTC 13443 | Detected | | | Citrobacter freundii | CCUG 418T | ampC (CMY-72) | Detected | | | Morganella morganii subsp. siboni | ATCC 51206 | ampC (DHA-4) / CDC 3522-75 | Detected | | | Morganella morganii | ATCC 25829 | ampC (DHA) / M4 | Detected | K262168 - Page 32 of 94 {32} | | K. pneumoniae | JMI Strain 1238756 | ampC (FOX-5) / N/A | Detected | | --- | --- | --- | --- | --- | | | E. coli | NCTC 14333 | ampC (CMY-LAT) / MLST: 167 | Detected | | | E. coli | NCTC 14339 | ampC (CMY-LAT) / MLST: 361 | Detected | | armA | A. baumannii | ATCC BAA-2803 | armA / 1125209 | Detected | | | A. baumannii | NCTC 13424 | armA / UK 20 | Detected | | | K. pneumoniae | ATCC BAA-3072 | armA / 979620 | Detected | | | A. baumannii | ATCC BAA-2873 | armA / 1081925 | Detected | | | K. pneumoniae | ATCC BAA-2783 | armA / 972863 | Detected | | | K. pneumoniae | CCUG 68728 | armA / NCTC 13443 | Detected | | | K. oxytoca | ATCC BAA-3059 | armA / 957532 | Detected | | CTX-M | E. coli | ATCC BAA-2774 | CTX-M-15 (Group 1) | Detected | | | E. coli | ATCC BAA-2775 | CTX-M-14 (Group 9) | Detected | | | K. pneumoniae | ATCC BAA-2783 | CTX-M-15 (Group 1) | Detected | | | K. pneumoniae | ATCC BAA-3077 | CTX-M-15 (Group 1) | Detected | | | gBlock | gBl…