MRSA/SA ELITE MGB
Device Facts
| Record ID | K112937 |
|---|---|
| Device Name | MRSA/SA ELITE MGB |
| Applicant | Elitech |
| Product Code | NQX · Microbiology |
| Decision Date | Jun 1, 2012 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 866.1640 |
| Device Class | Class 2 |
Intended Use
MRSA/SA ELITe MGB® is a qualitative in vitro diagnostic test for the direct detection of Staphylococcus aureus (SA) and methicillin-resistant Staphylococcus aureus (MRSA) using DNA purified from nasal swabs. MRSA/SA ELITe MGB® is intended to aid in the prevention and control of MRSA infections in healthcare settings. It is not intended to diagnose, guide or monitor MRSA infections, or provide results of susceptibility to oxacillin/methicillin. A negative result does not preclude MRSA/SA (Staphylococcus aureus) nasal colonization. Concomitant cultures are necessary to recover organisms for epidemiological typing or for further susceptibility testing.
Device Story
MRSA/SA ELITe MGB is a real-time, multiplex PCR assay for qualitative detection of MRSA and SA DNA from nasal swabs. Sample processing occurs on the bioMérieux NucliSENS easyMAG instrument. Extracted DNA is combined with monoreagent (primers, probes, master mix) and analyzed on the Applied Biosystems 7500 FAST Dx System. The system identifies the gene responsible for methicillin resistance (mecA) and a conserved S. aureus gene. Results are determined by an algorithm comparing Cq (Ct) values from the cycler. Used in clinical laboratories by trained personnel to aid in MRSA infection control. Output informs healthcare providers of colonization status; negative results do not preclude colonization, and concomitant cultures are required for epidemiological typing or susceptibility testing.
Clinical Evidence
Prospective study at 3 sites (n=3174 eligible specimens) compared the device to broth culture followed by latex agglutination and cefoxitin susceptibility testing. MRSA sensitivity: 92.3% (95% CI: 88.08-95.16%), specificity: 95.2% (95% CI: 94.32-95.87%). SA sensitivity: 96.1% (95% CI: 94.48-97.25%), specificity: 95.1% (95% CI: 94.16-95.89%). Discrepant analysis using SCCmec sequencing confirmed additional MRSA-positive cases.
Technological Characteristics
Real-time multiplex PCR assay. Targets: mecA gene and conserved S. aureus gene. Instrumentation: bioMérieux NucliSENS easyMAG (extraction) and Applied Biosystems 7500 FAST Dx (amplification/detection). Reagents: monoreagent containing primers, probes, and master mix. Qualitative detection based on Cq (Ct) values. Storage: -20°C.
Indications for Use
Indicated for qualitative detection of SA and MRSA DNA from nasal swabs in patients at risk for nasal colonization to aid in MRSA infection prevention and control in healthcare settings. Not for diagnosing, guiding, or monitoring MRSA infections or determining oxacillin/methicillin susceptibility.
Regulatory Classification
Identification
An antimicrobial susceptibility test powder is a device that consists of an antimicrobial drug powder packaged in vials in specified amounts and intended for use in clinical laboratories for determining in vitro susceptibility of bacterial pathogens to these therapeutic agents. Test results are used to determine the antimicrobial agent of choice in the treatment of bacterial diseases.
Predicate Devices
- BD GENEOHM MRSA ACP ASSAY (K093346)
Related Devices
- K132468 — MRSA/SA ELITE MGB, ELITE MGB SOFTWARE · Elitechgroup Epoch Biosciences · Oct 17, 2013
- K142721 — cobas MRSA/SA Test · Roche Molecular Systems, Inc. · Dec 17, 2014
Submission Summary (Full Text)
{0}------------------------------------------------
#### 510(k) Summary MRSA/SA ELITe MGB®
substantial equivalence.
The assigned 510(k) number is: K112937
| Submitter:<br>Address:<br>Phone number<br>Fax number | ELITechGroup Epoch Biosciences.<br>21720 23rd Dr SE, Suite 150, Bothell, WA 98021 USA<br>425-482-5555<br>425-482-5550 |
|------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------|
| Contact | Debra K Hutson (Email: d.hutson@elitechgroup.com) |
| Date of Preparation | September 29, 2011 |
#### Device:
Introduction
| Trade/proprietary Name:. | MRSA/SA ELITE MGB® |
|---------------------------------------|------------------------------------------------------------------------------------------------------------|
| Common or Usual Name: | Test Kit for the detection of methicillin-resistant Staphylococcus aureus |
| Regulation number/name: | 866.1640, Antimicrobial susceptibility test powder |
| Device Class: | Class II |
| Product code | NQX |
| Classification name | System, nucleic acid amplification test, dna, methicillin resistant staphylococcus aureus, direct specimen |
| Classification Advisory<br>Committee: | Microbiology |
| Panel: | 83 |
| Predicate device | BD GENEOHM MBSA ACP ASSAY (K093346) |
Device description: MRSA/SA ELITe MGB is a real-time, multiplex polymerase chain reaction (PCR) assay for the in vitro qualitative detection of MRSA and SA DNA extracted from human nasal swab samples. In this system, sample preparation and amplification/real-time detection are completed on separate instruments.
Sample processing is completed on the bioMérieux NucliSENS® easyMAG® instrument with bioMérieux NucliSENS Nucleic Acid Extraction Reagents according to the manufacturer's instructions. Following processing, the extracted sample is placed in the well of a 96 well plate to which "monoreagent" is added. The monoreagent contains the primers and probes for the genes of interest and the internal control combined with master mix. The assay is performed on an Applied Biosystems 7500 FAST Dx System that consists of the 7500 FAST Dx instrument, a personal computer, 96-well plates and seals. The total system run time is 150 minutes consisting of 60 minutes for sample processing and about 90 minutes for the real time amplification and detection steps. The instrument never comes into contact with any fluids within the 96well plate. Each disposable plate is intended to test up to 96 samples, controls or any mixture thereof. The 96-well plates are not re-usable and are specific to the system. The kit contains enough reagents for 100 reactions. One positive and one negative control are required for each PCR run ; a Negative
{1}------------------------------------------------
| | Processing Control and a Positive Processing Control are recommended to be<br>run in each extraction run. The design of the assay includes systems to identify<br>both the gene responsible for methicillin resistance and for a conserved portion<br>of a gene unique to S. aureus. Thus, for a true "MRSA," both targets will be<br>identified in roughly equal proportions. Results are determined by using an<br>algorithm that compares output, Cq, from the cycler (called Ct in the output from<br>the cycler.) |
|---------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use: | MRSA/SA ELITe MGB® is a qualitative in vitro diagnostic test for the direct<br>detection of Staphylococcus aureus (SA) and methicillin-resistant<br>Staphylococcus aureus (MRSA) using DNA purified from nasal swabs.<br>MRSA/SA ELITe MGB® is intended to aid in the prevention and control of<br>MRSA infections in healthcare settings. It is not intended to diagnose, guide or<br>monitor MRSA infections, or provide results of susceptibility to<br>oxacillin/methicillin. A negative result does not preclude MRSA/SA<br>(Staphylococcus aureus) nasal colonization. Concomitant cultures are<br>necessary to recover organisms for epidemiological typing or for further<br>susceptibility testing. |
| Indication for use: | MRSA/SA ELITe MGB® is a qualitative in vitro diagnostic test for the direct<br>detection of Staphylococcus aureus (SA) and methicillin-resistant<br>Staphylococcus aureus (MRSA) using DNA purified from nasal swabs.<br>MRSA/SA ELITe MGB® is intended to aid in the prevention and control of<br>MRSA infections in healthcare settings. It is not intended to diagnose, guide or<br>monitor MRSA infections, or provide results of susceptibility to<br>oxacillin/methicillin. A negative result does not preclude MRSA/SA<br>(Staphylococcus aureus) nasal colonization. Concomitant cultures are<br>necessary to recover organisms for epidemiological typing or for further<br>susceptibility testing. |
## Comparison to Predicate device
| | ELITech Molecular Diagnostics Device<br>MRSA/SA ELITe MBG | Predicate device<br>BD GENEOHM MRSA ACP ASSAY<br>(K093346) |
|-------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended use | MRSA/SA ELITe MGB® is a qualitative<br>in vitro diagnostic test for the direct<br>detection of Staphylococcus aureus<br>(SA) and methicillin-resistant<br>Staphylococcus aureus (MRSA) using<br>DNA purified from nasal swabs.<br>MRSA/SA ELITe MGB® is intended to<br>aid in the prevention and control of<br>MRSA infections in healthcare<br>settings. It is not intended to diagnose,<br>guide or monitor MRSA infections, or<br>provide results of susceptibility to<br>oxacillin/methicillin. A negative result<br>does not preclude MRSA/SA<br>(Staphylococcus aureus) nasal<br>colonization. Concomitant cultures are<br>necessary to recover organisms for<br>epidemiological typing or for further | The BD GeneOhm® MRSA ACP<br>Assay is a qualitative in vitro<br>diagnostic test for the direct detection<br>of methicillin-resistant Staphylococcus<br>aureus (MRSA) DNA from nasal<br>swabs in patients at risk for nasal<br>colonization. The test utilizes<br>polymerase chain reaction (PCR) for<br>the amplification of MRSA DNA and<br>fluorogenic target-specific<br>hybridization probes for the detection<br>of the amplified DNA. The BD<br>GeneOhm® MRSA ACP Assay is<br>intended to aid in the prevention and<br>control of MRSA infections in<br>healthcare settings. It is not intended<br>to diagnose MRSA infections nor to<br>guide or monitor treatment for MRSA |
| | ELITech Molecular Diagnostics Device | Predicate device |
| | MRSA/SA ELITE MBG® | BD GENEOHM MRSA ACP ASSAY<br>(K093346) |
| | susceptibility testing. | infections.<br>Concomitant cultures are necessary<br>only to recover organisms for<br>epidemiological typing or for further<br>susceptibility testing. |
| Indication for Use | MRSA/SA ELITE MGB® is a qualitative<br>in vitro diagnostic test for the direct<br>detection of Staphylococcus aureus<br>(SA) and methicillin-resistant<br>Staphylococcus aureus (MRSA) using<br>DNA purified from nasal swabs.<br>MRSA/SA ELITE MGB® is intended to<br>aid in the prevention and control of<br>MRSA infections in healthcare<br>settings. It is not intended to diagnose,<br>guide or monitor MRSA infections, or<br>provide results of susceptibility to<br>oxacillin/methicillin. A negative result<br>does not preclude MRSA/SA<br>(Staphylococcus aureus) nasal<br>colonization. Concomitant cultures are<br>necessary to recover organisms for The BD GeneOhm® MRSA ACP<br>Assay is a qualitative <i>in vitro</i><br>diagnostic test for the direct detection<br>of methicillin-resistant <i>Staphylococcus</i><br><i>aureus</i> (MRSA) DNA from nasal<br>swabs in patients at risk for nasal<br>colonization. The test utilizes<br>polymerase chain reaction (PCR) for<br>the amplification of MRSA DNA and<br>fluorogenic target-specific<br>hybridization probes for the detection<br>of the amplified DNA. The BD<br>GeneOhm® MRSA ACP Assay is<br>intended to aid in the prevention and<br>control of MRSA infections in<br>healthcare settings. It is not intended<br>to diagnose MRSA infections nor to<br>guide or monitor treatment for MRSA<br>infections.<br>Concomitant cultures are necessary<br>only to recover organisms for<br>epidemiological typing or for further<br>susceptibility testing. | |
| Mode of identification<br>of <i>S. aureus</i> | Presence of conserved region in a<br><i>Staphylococcus aureus</i> -specific gene. | Presence of SCC <i>mec</i> cassette<br>(genetic element that carries the <i>mecA</i><br>gene) at <i>orfX</i> junction (specific to <i>S.</i><br><i>aureus</i> ) |
| Mode of detection for<br>methicillin resistance | Presence of the <i>mecA</i> gene which is<br>responsible for resistance to<br>methicillin. | gene) at <i>orfX</i> junction (specific to <i>S.</i><br><i>aureus</i> ) |
| Assay Format | Qualitative real-time polymerase chain<br>reaction (PCR) assay using 3 forward<br>primer, 3 reverse<br>primers, and 3 fluorescent-labeled<br>probes for the amplification and<br>detection of methicillin resistant<br><i>Staphylococcus aureus</i> (MRSA) DNA. | Qualitative real-time polymerase chain<br>reaction (PCR) assay using 1 forward<br>primer, 5 reverse<br>primers, and 2 molecular beacon<br>probes for the amplification and<br>detection of methicillin resistant<br><i>Staphylococcus aureus</i> (MRSA) DNA. |
| Composition | MRSA/SA ELITE MGB® PCR Mix<br>Tfi PCR Master Mix<br><0.01% MRSA/SA primers<br><0.01% Internal Control primers<br><0.01% MRSA/SA Fluorescent-<br>labeled oligonucleotide probes<br><0.01% Internal Control Fluorescent-<br>labeled oligonucleotide probe | Master Mix<br>< 0.0005% DNA polymerase complex<br>< 0.001% Internal Control: non-<br>infectious DNA containing MRSA-<br>primer binding sequences and a<br>unique sequence for probe<br>hybridization<br>< 0.06% primers |
| | ELITech Molecular Diagnostics Device<br>MRSA/SA ELITE MBG® | Predicate device<br>BD GENEOHM MRSA ACP ASSAY<br>(K093346) |
| | Reference dT(8)-AP593<br>MRSA/SA Internal Control<br>Tris buffer<br><0.01% EDTA<br>0.01% total yeast RNA<br><0.001% Non-infectious plasmid DNA<br>(recombinant) containing Internal<br>Control sequences | < 1% Nucleotide mix (dATP, dCTP,<br>dGTP, dTTP)<br>Bovine serum albumin<br>Carbohydrate<br>MgCl2<br>< 0.001% non-infectious<br>Staphylococcus epidermidis genomic<br>DNA (ATCC 14990) |
| | MRSA/SA Positive Control<br>Tris buffer<br><0.01% EDTA<br>0.01% total yeast RNA<br><0.001% Non-infectious plasmid DNA<br>(microbial) containing MRSA<br>sequences | Control DNA<br>Tris-EDTA buffer<br>Carbohydrate<br>< 0.001% non-infectious genomic<br>MRSA DNA (ATCC 43300)<br>Diluent<br>Tris-HCl buffer<br>MgCl2<br>(NH4)2SO4<br>KCl<br>Tween-20 |
| Appearance of<br>reagents | Frozen, ready to use | Liquid, ready to use. |
| Sample type | Nasal swab | Nasal swab |
| Storage & Expiry | Stored in -20 °C freezer. The device<br>is stable until the expiry date stated on<br>the label. | Stored at 2- 25 °C.<br>Reagents are stable until the expiry<br>date stated on the label. |
| Instrument | ABI 7500 Fast Dx | BD SmartCycler® II |
| Controls | Positive PCR control (Plasmid DNA<br>(microbial) containing MRSA<br>sequences)<br>Internal Control (Plasmid DNA<br>(recombinant) containing Internal<br>Control sequences) | Positive PCR control (DNA from S.<br>aureus ATCC 43300).<br>Negative PCR control (DNA from S.<br>epidermidis ATCC 14990).<br>Internal procedural control |
:
:
{2}------------------------------------------------
.
.
1. 1. 1. 1. 1.
:
{3}------------------------------------------------
## Standard/Guidance Document Referenced
FDA Draft Guidance for Industry and Food and Drug Administration Staff Establishing the Performance Characteristics of Nucleic Acid-Based In vitro Diagnostic Devices for the Detection and Differentiation of Methicillin-Resistant Staphylococus aureus (MRSA) and Staphylococcus aureus (SA), Issued January 5, 2011.
FDA Guidance for Industry, FDA Reviewers and Compliance on Off-the-Shelf Software Use in Medical Devices, Issued September 9, 1999.
{4}------------------------------------------------
## Instrumentation/Software:
The system is performed with FDA-cleared devices, bioMérieux NucliSENS® easyMAG® extraction system and the Applied Biosystems 7500 Fast Dx PCR Instrument. ELITechGroup Epoch Biosciences has a relationship with each on the manufacturers of these devices via service contracts such that Epoch will become aware, in the same time, as other users of the system, of changes to the device(s) or of the software used by the device(s). Internal quality assurance procedures are in place to verify the continued acceptable performance of the test device. Please, note, however, that the evaluation algorithm and the use of controls as indicated in the labeling, Internal Control and Positive Control, Negative Specimen Processing Control and Positive Specimen Processing Control, should identify for users any issues created by instrument or software changes.
#### Performance Characteristics
#### Interfering substances
A study was conducted with potentially interfering substances encountered on nasal swabs. Substances tests were chemical substances that can either be naturally present in the nasal cavity or that can be artificially introduced into the nasal cavity.
The following substances were tested and evaluated: blood, mucin, phenylephrine (Neo-synephrine®), oxymetazoline (Dristan®, Zicam®), sodium chloride with preservatives, benzalkonium chloride, sodium phosphate, phenylcarbinol (Saline), propylene glycol (AYR® saline nasal gel), sorbitol, benzyl alcohol, disodium EDTA, hypromellose, phosphoric acid, dexamethasone, triamcinolone (Nasacort"), beclomethasone (Beconase AQ"), flunisolide, budesonide (Rhinocort Aqua"), mometasone (Nasonex"), fluticasone (Flonase®), luffa opperculata, sulfur, Galphimia glauca, Histaminum hydrochloricum, live intranasal influenza virus vaccine (FluMist®), benzocaine, methol (Cepacol® sore throat lozenges), Zanamivir (Relenza"), Oseltamivir phosphate (Tamiflu©), Mupirocin, tobramycin.
The following substances have been shown to interfere with the performance of the assay: AYFO saline nasal gel and excessive amounts of nasal secretions/mucus.
## Non-Clinical Performance Evaluation
#### A. Analytical Sensitivity
The analytical sensitivity of the MRSA/SA ELITe MGB was determined using 5 strains of MRSA and one MSSA strain. Cultures of these strains were quantified, diluted in simulated nasal matrix to values spanning the range of approximately 5 to 1500 colonies forming units (CFU) and absorbed onto swabs. All dilutions were tested, and the limit of detection (LoD) was determined by Probit analysis. LoD for each strain represents the lowest number of CFU/swab at which a positive result will be obtained with at least 95% confidence. LoD for each strain was then verified by testing at least 20 replicates. Results indicate that the MRSA/SA ELITe MGB® average LoD is 165 CFU/mL of a swab eluate.
#### B. Analytical Reactivity
#### Inclusivity
Performance of the MRSA/SA ELITe MGB® was tested on 75 well characterized MRSA and MSSA isolates representative of the qlobal genetic diversity, including clonal complexes and sequence types as well as various Pulse-Field Gel Electrophoresis (PFGE) types and MIC (Minimum Inhibitory Concentration) values, with the emphasis on the USA enidemiologic clones. The strains were obtained through the Network on Antimicrobial Resistance in Staphylococcus aureus (NARSA) Program and from American Tissue Culture Collection (ATCC) or were a gift from Medical College of Wisconsin'. All strains were absorbed onto swabs at near detection limit and tested with MRSA/SA ELITe MGB. In addition to that all
<sup>1</sup> Gift from Dr. Nathan A. Ledeboer, Medical College of Wisconsin, Wi; the strains are described in: Buchan, B.W, Ledeboer. N.A. Identification of Two Borderline Oxacillin-Resistant Strains of Staphylococcus aureus From Routine Nares Swab Specimens by One of Three Chromogenic Agars Evaluated for the Detection of MRSA, Microbiology and Infectious Disease.2010:134;921-927.
{5}------------------------------------------------
MSSA strains were tested at 10 CFU/swab. All MSSA strains tested positive for SA and negative for MRSA. All MRSA strains tested positive for MRSA. Two BORSA (Borderline Oxacillin Resistant Staphylococcus aureus) isolates that lack mecA 9 tested SA positive and MRSA negative for an overall analytical reactivity of 97.3%.
#### ். Analytical Specificity
## Exclusivity
The specificity of the MRSA/SA ELITe MGB® was evaluated by testing for cross-reactivity to species phylogenetically related to S. aureus, pathogenic microorganisms and to microorganisms commonly present in normal nasal microflora. The test panel consisted of 17 viral, 1 mycoplasma, and 41 bacterial species. The microorganisms were tested as cultures in concentrations of 1x10" CFU (1x10" PFU)/swab. In addition human cells in a concentration of 10° cells /mL were tested. Human cells and all tested species were found negative for MRSA and SA with the MRSA/SA ELITE MGB®. The analytical specificity was 100%.
| Staphylococci Species | Other Organisms | Viruses |
|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| CoNS*<br>Staphylococcus arlettae,<br>Staphylococcus capitis,<br>Staphylococcus carnosus,<br>Staphylococcus chromogenes,<br>Staphylococcus equorum,<br>Staphylococcus felis,<br>Staphylococcus gallinarum,<br>Staphylococcus hominis<br>subsp. hominis,<br>Staphylococcus kloosii,<br>Staphylococcus lentus,<br>Staphylococcus pulvereri,<br>Staphylococcus simulans,<br>Staphylococcus warneri<br>MSCoPS*<br>Staphylococcus delphini,<br>MSCoNS*<br>Staphylococcus epidermidis,<br>Staphylococcus xylosus<br>MRCONS*<br>Staphylococcus epidermidis<br>CoPS*<br>Staphylococcus hyicus,<br>Staphylococcus intermedius | Acinetobacter haemolyticus, Bacillus<br>cereus, Bordetella pertussis,<br>Citrobacter freundii, Citrobacter koseri,<br>Corynebacterium aquaticum,<br>Corynebacterium bovis,<br>Corynebacterium flavescens,<br>Corynebacterium genitalium,<br>Enterobacter aerogenes,<br>Enterococcus faecalis, Enterococcus<br>faecium, Enterococcus flavescens,<br>Enterococcus gallinarum,<br>Enterococcus hiraem, Escherichia coli,<br>ESBL producer, Klebsiella oxytoca,<br>Klebsiella pneumoniae, ESBL<br>producer, Listeria monocytogenes,<br>Moraxella catarrhalis, Pasteurella<br>aerogenes, Proteus mirabilis, Proteus<br>vulgaris, Pseudomonas aeruginosa,<br>Salmonella typhimurium, Serratia<br>marcescens, Shigella sonnei,<br>Streptococcus mitis, Streptococcus<br>salivarius, Yersinia enterocolitica,<br>Candida albicans, Candida glabrata,<br>Cryptococcus neoformans,<br>Lactobacillus acidophilus, Legionella<br>pneumophila, Mycobacterium<br>tuberculosis avirulent, Mycoplasma<br>pneumoniae, Neisseria meningitides,<br>Streptococcus mutans, Streptococcus<br>pneumoniae, Streptococcus<br>pyogenes, Homo sapiens, Human<br>Cells HT1080 | Adenovirus Type 1,<br>Adenovirus Type 7A,<br>Human coronavirus (229E),<br>Human coronavirus (OC43),<br>Cytomegalovirus,<br>Coxsackievirus Type A21,<br>Epstein Barr Virus,<br>Human influenza virus A,<br>Human influenza virus B,<br>Human parainfluenza Type 2,<br>Human parainfluenza Type 3,<br>Human metapneumovirus 3 Type<br>B1,<br>Measles,<br>Mumps virus,<br>Respiratory syncytial virus Type B,<br>Rhinovirus Type 1A |
## Species Tested for Cross-Reactivity and Microbial Interference
* CoNS: coagulase-negative Staphylococci, MSCoPS: methicillin susceptible coagulase positive Staphylococci, MSCoNS: methicillin susceptible coagulase negative Staphylococci,
{6}------------------------------------------------
MRCoNS: methicillin resistant coaqulase negative Staphylococci. CoPS: coaqulase positive Staphylococci
Two of the potentially interfering organisms tested, Human metapneumovirus (hMPV) and MRCoNS Staphylococcus epidermidis, strain NRS 34, caused interference at initial test.
Re-testing of MRCoNS Staphylococus epidermidis, strain NRS 34 confirmed the interference results. MRCoNS Staphylococcus epidermidis, strain NRS 34, is considered to interfere with MRSA/SA ELITE MGB®.
- MRCoNS Staphylococcus epidermidis when tested in a mix with near-detection-limit -MRSA resulted in "SA positive, MRSA negative" call.
Re-testing of Human metapneumovirus (hMPV) did not reveal interference of hMPV with MRSA/SA ELITe MGB":
Methicillin Susceptible S. aureus (MSSA) was also tested for microbial interference. The microorganism was spiked at 1×10° CFU/mL (1×10° PFU/mL), or higher, into a sample with MRSA strains at neardetection-limit and tested.
- MRCoNS Staphylococcus epidermidis and MSSA, when tested in a mix with near-detection-. limit of MRSA, resulted in "SA positive, MRSA neqative" call.
None of the other tested species interfered with MRSA/SA detection.
#### D. Reproducibility
.
A 10-member panel of specimens with varying concentrations of MRSA in a simulated nasal matrix was tested. Two MRSA strains (ATCC BAA-1720) and one MSSA strain (BAA-12600) were used. Simulated matrix contained human genomic DNA and mucin to imitate a normal human nasal matrix. Data from the specimens were pooled to produce four sample types: For each MRSA/MSSA strain the panel included a negative member, specimen below the LoD (expected to yield a positivity rate of between 20 to 80%), low positive (at LoD, expected to yield a 95% positivity rate), and moderate positive (three times LoD, expected to have 100% positivity rate).
Each of the two operators performed one run per day for 12 days on three reagent lots at one site. In two other sites two runs per day on one reagent lot were performed for 5 days (10 specimens x 3 replicates x 5 days x 2 runs).
The negative panel member yielded negative results 100%, the below LoD specimens positivity rate was 77%, the low positive specimen positivity rate was 98%, and the moderate positive panel members positivity rate was 100%.
| Specimen Type | Lot 1 | Lot 2 | Lot 3 | Total Agreement (%) |
|-------------------------------|-------|-------|-------|---------------------|
| Negative (R1) | 14/14 | 14/14 | 30/30 | 58/58 (100%) |
| Below LoD (R2,R5,R8) | 33/42 | 31/42 | 70/90 | 134/174 (77%) |
| Low Positive (R3,R6,R9) | 40/42 | 42/42 | 88/90 | 170/174 (98%) |
| Moderate Positive (R4,R7,R10) | 42/42 | 42/42 | 90/90 | 174/174 (100%) |
#### Cumulative data of reproducibility study
The numerical results based on Ct values follow:
{7}------------------------------------------------
| 4 | |
|---|--|
| 1 | |
| | |
| | |
| | |
| | |
| | |
| | |
| | |
| | |
| | |
| | |
| | Panel # | N | Mean Ct | Within-Run | | Between-Run | | Between-Day | | Between-Operator | | Between-Lot | | Between-System | | Total | |
|------|---------|-------|---------|------------|------|-------------|------|-------------|------|------------------|------|-------------|------|----------------|------|-------|-----|
| | | | | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% |
| R1 | 118 | 38.12 | NA1 | NA1 | 0.67 | 1.75 | 0.45 | 1.18 | 0.29 | 0.76 | 0.39 | 1.03 | 0.15 | 0.40 | 0.39 | 1.02 | |
| R2 | 118 | 36.22 | 0.65 | 1.79 | 0.99 | 2.74 | 0.79 | 2.18 | 0.82 | 2.26 | 0.32 | 0.89 | 0.86 | 2.38 | 0.74 | 2.04 | |
| R3 | 118 | 35.39 | 0.62 | 1.73 | 0.96 | 2.72 | 0.86 | 2.42 | 0.94 | 2.65 | 0.43 | 1.23 | 1.08 | 3.06 | 0.82 | 2.30 | |
| R4 | 118 | 34.35 | 0.41 | 1.20 | 0.67 | 1.94 | 0.53 | 1.54 | 0.42 | 1.22 | 0.48 | 1.40 | 0.35 | 1.03 | 0.48 | 1.39 | |
| R5 | 118 | 36.73 | 0.71 | 1.94 | 0.87 | 2.37 | 0.79 | 2.16 | 0.56 | 1.52 | 0.50 | 1.37 | 0.45 | 1.23 | 0.65 | 1.77 | |
| R6 | 118 | 33.66 | 0.42 | 1.24 | 0.69 | 2.05 | 0.64 | 1.92 | 0.56 | 1.66 | 0.48 | 1.44 | 0.58 | 1.72 | 0.56 | 1.67 | |
| R7 | 118 | 31.81 | 0.17 | 0.54 | 0.76 | 2.38 | 0.67 | 2.09 | 0.73 | 2.28 | 0.43 | 1.36 | 0.83 | 2.60 | 0.60 | 1.88 | |
| R8 | 118 | 37.68 | 0.77 | 2.05 | 0.74 | 1.97 | 0.43 | 1.15 | 0.39 | 1.03 | 0.12 | 0.33 | 0.40 | 1.06 | 0.48 | 1.26 | |
| R9 | 118 | 34.65 | 0.56 | 1.60 | 0.64 | 1.84 | 0.51 | 1.49 | 0.54 | 1.56 | 0.08 | 0.23 | 0.60 | 1.74 | 0.49 | 1.41 | |
| R10 | 118 | 32.82 | 0.58 | 1.76 | 0.48 | 1.45 | 0.42 | 1.28 | 0.37 | 1.13 | 0.17 | 0.53 | 0.40 | 1.21 | 0.40 | 1.23 | |
| PSPC | 44 | 34.25 | NA2 | NA2 | 0.71 | 2.08 | 0.41 | 1.21 | 0.41 | 1.18 | 0.18 | 0.52 | 0.59 | 1.72 | 0.46 | 1.34 | |
| PC | 44 | 28.12 | NA2 | NA2 | 1.08 | 3.86 | 1.18 | 4.20 | 0.58 | 2.07 | 1.21 | 4.24 | 0.50 | 1.79 | 0.91 | 3.23 | |
| NSPC | 44 | 38.24 | NA2 | NA2 | 0.63 | 1.64 | 0.65 | 1.69 | 0.85 | 2.24 | 0.11 | 0.30 | 0.82 | 2.15 | 0.61 | 1.60 | |
| NTC | 44 | 38.14 | NA3 | NA3 | NA3 | NA3 | NA3 | NA3 | NA3 | NA3 | NA3 | NA3 | NA3 | NA3 | NA3 | NA3 | |
mecA
| N | Mean<br>Ct | Within-Run | | Between-Run | | Between-Day | | Between-Operator | | Between-Lot | | Between-System | | Total | |
|-----|------------|------------|------|-------------|------|-------------|------|------------------|------|-------------|------|----------------|------|-------|------|
| | | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% |
| 118 | 38.53 | NA1 | NA1 | 1.08 | 2.81 | 1.31 | 3.42 | 0.76 | 1.97 | 0.97 | 2.54 | 0.32 | 0.83 | 0.89 | 2.31 |
| 118 | 36.87 | 0.53 | 1.43 | 0.93 | 2.52 | 0.68 | 1.84 | 0.68 | 1.85 | 0.40 | 1.09 | 0.74 | 2.02 | 0.66 | 1.79 |
| 118 | 36.40 | 0.59 | 1.62 | 1.13 | 3.12 | 0.99 | 2.73 | 1.15 | 3.15 | 0.66 | 1.83 | 1.27 | 3.51 | 0.97 | 2.66 |
| 118 | 35.17 | 0.37 | 1.05 | 0.68 | 1.94 | 0.54 | 1.54 | 0.46 | 1.32 | 0.43 | 1.24 | 0.41 | 1.17 | 0.48 | 1.38 |
| 118 | 37.31 | 0.60 | 1.62 | 0.64 | 1.71 | 0.50 | 1.35 | 0.44 | 1.17 | 0.47 | 1.26 | 0.46 | 1.24 | 0.52 | 1.39 |
| 118 | 34.45 | 0.43 | 1.24 | 0.74 | 2.15 | 0.70 | 2.04 | 0.58 | 1.68 | 0.61 | 1.78 | 0.60 | 1.74 | 0.61 | 1.77 |
| 118 | 32.50 | 0.19 | 0.58 | 0.87 | 2.68 | 0.76 | 2.34 | 0.85 | 2.61 | 0.52 | 1.61 | 0.95 | 2.91 | 0.69 | 2.12 |
| 118 | 39.12 | NA4 | NA4 | 0.55 | 1.42 | 0.52 | 1.35 | 0.45 | 1.16 | 0.25 | 0.65 | 0.33 | 0.86 | 0.42 | 1.09 |
| 118 | 38.83 | NA4 | NA4 | 0.62 | 1.59 | 0.46 | 1.17 | 0.38 | 0.97 | 0.14 | 0.35 | 0.32 | 0.83 | 0.38 | 0.98 |
| 118 | 38.74 | NA4 | NA4 | 0.77 | 1.98 | 0.57 | 1.46 | 0.40 | 1.05 | 0.36 | 0.94 | 0.31 | 0.79 | 0.48 | 1.25 |
| 44 | 35.03 | NA2 | NA2 | 0.73 | 2.08 | 0.56 | 1.60 | 0.42 | 1.20 | 0.28 | 0.80 | 0.56 | 1.60 | 0.51 | 1.45 |
| 44 | 29.18 | NA2 | NA2 | 1.11 | 3.82 | 1.23 | 4.21 | 0.51 | 1.74 | 1.24 | 4.22 | 0.40 | 1.37 | 0.90 | 3.07 |
| 44 | 39.03 | NA2 | NA2 | 0.48 | 1.22 | 0.48 | 1.22 | 0.48 | 1.22 | 0.58 | 1.48 | NA5 | NA5 | 0.50 | 1.28 |
| 44 | 39.53 | NA2 | NA2 | 0.24 | 0.61 | 0.24 | 0.61 | 0.24 | 0.61 | NA6 | NA6 | 0.24 | 0.61 | 0.24 | 0.61 |
ﻓﮩﺮﺳ ﺑﮭﺎﺭﺗﯽ
ﺮ ﭘﻮﺭ ﺍﻭ ﭘﻪ ﭘﻪ
Page 5.8 of 5.11 pages
ম
ার্টি
ﻦ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺪﻳﻨﺔ
{8}------------------------------------------------
| C2 | Panel<br># | N | Mean<br>Ct | Within-Run | | Between-Run | | Between-Day | | Between-Operator | | Between-Lot | | Between-System | | Total | |
|----|------------|-----|------------|------------|------|-------------|------|-------------|------|------------------|------|-------------|------|----------------|------|-------|------|
| | | | | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% |
| | R1 | 118 | 30.26 | 0.15 | 0.50 | 0.27 | 0.89 | 0.17 | 0.55 | 0.18 | 0.59 | 0.05 | 0.18 | 0.23 | 0.75 | 0.17 | 0.58 |
| | R2 | 118 | 30.24 | 0.18 | 0.58 | 0.30 | 1.00 | 0.24 | 0.79 | 0.18 | 0.60 | 0.03 | 0.10 | 0.24 | 0.78 | 0.19 | 0.64 |
| | R3 | 118 | 30.30 | 0.17 | 0.57 | 0.25 | 0.81 | 0.18 | 0.60 | 0.17 | 0.57 | 0.04 | 0.14 | 0.20 | 0.66 | 0.17 | 0.56 |
| | R4 | 118 | 30.26 | 0.12 | 0.39 | 0.27 | 0.88 | 0.16 | 0.53 | 0.17 | 0.57 | 0.11 | 0.36 | 0.18 | 0.58 | 0.17 | 0.55 |
| | R5 | 118 | 30.23 | 0.22 | 0.72 | 0.33 | 1.09 | 0.24 | 0.78 | 0.23 | 0.75 | 0.10 | 0.34 | 0.27 | 0.88 | 0.23 | 0.76 |
| | R6 | 118 | 30.38 | 0.22 | 0.74 | 0.43 | 1.40 | 0.31 | 1.01 | 0.14 | 0.47 | 0.06 | 0.20 | 0.09 | 0.30 | 0.21 | 0.69 |
| | R7 | 118 | 30.31 | 0.15 | 0.50 | 0.30 | 0.98 | 0.21 | 0.68 | 0.13 | 0.42 | 0.04 | 0.12 | 0.14 | 0.45 | 0.16 | 0.53 |
| | R8 | 118 | 30.39 | 0.23 | 0.76 | 0.38 | 1.24 | 0.24 | 0.79 | 0.25 | 0.81 | 0.21 | 0.68 | 0.20 | 0.66 | 0.25 | 0.82 |
| | R9 | 118 | 30.34 | 0.14 | 0.46 | 0.32 | 1.06 | 0.23 | 0.74 | 0.28 | 0.93 | 0.12 | 0.38 | 0.26 | 0.87 | 0.23 | 0.74 |
| | R10 | 118 | 30.16 | 0.24 | 0.80 | 0.51 | 1.70 | 0.27 | 0.89 | 0.31 | 1.04 | 0.13 | 0.42 | 0.33 | 1.09 | 0.30 | 0.99 |
| | PSPC | 44 | 30.26 | NA2 | NA2 | 0.47 | 1.55 | 0.36 | 1.18 | 0.32 | 1.05 | 0.33 | 1.09 | 0.28 | 0.94 | 0.35 | 1.16 |
| | PC | 44 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 |
| | NSPC | 44 | 30.27 | NA2 | NA2 | 0.63 | 2.09 | 0.51 | 1.68 | 0.48 | 1.59 | 0.35 | 1.15 | 0.32 | 1.05 | 0.46 | 1.51 |
| | NTC | 44 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 | NA7 |
- r I (Neglive Samples vols in triplicats show a waks ignal in the therer. Therefors, calculous of withi
S Dand CV (for the CNS) is not medialer as sed for estable o calcular
-
-
- in R-R10 samples.
in R-R10 samples (MSSA), due to none or rare (one per triplicate) occurrence of mecA signal detection it was not possible to calculate S
or this target.
- or no targer, por mobile princes per minene per rate signi a mains passo or anoto or anoto or ancolor de arcame or acades or ar
Por relizanes, and resident necessario necessa
-
-
{9}------------------------------------------------
#### Carry-Over / Cross-Contamination ய்
An analytical study was performed to evaluate the potential for cross-contamination between high MRSA (1x10' CFU per mL) specimens and negative specimens throughout the MRSA/SA ELITe MGB" workflow. Two operators performed five 24-sample extraction runs (11 high MRSA samples, 1 PSPC sample, and 1 NSPC sample per run) in a checkerboard pattern (high MRSA samples interrupted by completely negative samples). The processed samples were then PCR amplified in five separate runs using two different checkerboard patterns. The cross-contamination testing resulted in zero false neqatives from fifty-five high MRSA positive samples and one false positive sample from fifty-five negative samples.
#### Clinical Performance
Performance characteristics of the MRSA/SA ELITe MGB® were determined in a prospective investigational study at three (3) sites by comparing the MRSA/SA ELITe MGB® with agglutination/susceptibility tests. Specimens were collected from three (3) unique geographies at institutions having MRSA culture-based screening programs in place. To be enrolled in the study, patients had to be eligible for MRSA screening to the policies of the respective facilities. A true MRSA culture-positive specimen was defined as a specimen where MRSA was identified by the latex agglutination and cefoxitin susceptibility test after broth enrichment (both positive). A true MSSA culturepositive specimen was defined as a specimen negative for cefoxitin susceptibility testing and positive for the latex agglutination test.
One nasal swab was collected from each patient and used to inoculate a selective chromogenic MRSA screening agar plate. Then the swab was inserted into a tube with trypticase soy broth and thoroughly mixed. The entire volume of the cell suspension was tested using MRSA/SA ELITe MGB. All swabs were subjected to enrichment in trypticase soy broth with 6.5% NaCl. The enriched culture samples were inoculated onto Trypticase Soy Blood Agar plates. Grown overnight cultures were used for latex agglutination. Specimens positive for latex agglutination were used for the cefoxitin susceptibility test.
Performance of the MRSA/SA ELITe MGB® was calculated relative to the broth culture followed by latex agglutination and cefoxitin susceptibility test results.
A total of 3271 nasal swab specimens were collected and tested. Of the 3271 specimen tested, 3174 specimens were eligible to be included in statistical analyses: 72 specimens were considered to be ineligible due to a duplicating error during samples collection and preparation; 21 specimens failed the initial extraction due to an operator error. Those samples were retested from the original swabs. 25 specimens failed the extraction and have been removed from the study.
Compared to the reference culture method, MRSA/SA ELITe MGB® identified 92% of the specimens testing positive for MRSA and 95% of the negative specimens.
Compared to the reference culture method. MRSA/SA ELITe MGB® identified 96% of the specimens testing positive for SA and 95% of the negative specimens.
{10}------------------------------------------------
| Combined Data | | Reference Culture | | | |
|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------|-------------------|-----------|---------------|-------|
| | | MRSA+ | SA+/MRSA- | Neg/No Growth | Total |
| MRSA/SA ELITe<br>MGB® | MRSA+ | 205 | 111 | 32 | 348 |
| | SA+/MRSA- | 17 | 405 | 86 | 508 |
| | SA- | 0 | 30 | 2288 | 2318 |
|…
