PROGASTRO SSCS ASSAY

{0}------------------------------------------------ 123274 Page 1 of 11 01/15/2013 Gen-Probe Prodesse, Inc. ProGastro Assay 510(k) Submission ### Attachment D 510(k) SUMMARY ### CONTACT Karen Harrington, PhD Manager, Clinical Affairs Gen-Probe Prodesse. Inc. 20925 Crossroads Circle Waukesha, WI 53186 ### NAME OF DEVICE Trade Name: Regulation Number: Product Code: ProGastro SSCS Assay 21 CFR 866.3990 PCH and PCI ### PREDICATE DEVICE K121454 Luminex GPP Test #### INTENDED USE The Prodesse® ProGastro SSCS Assav is a multiplex real time PCR in vitro diagnostic test for the qualitative detection and differentiation of Salmonella, Shigella, and Campylobacter (C. jejimi and C. coli only, undifferentiated) nucleic acids and Shiga Toxin 1 (stxl) and Shiga Toxin 2 (stx2) genes. Shiga toxin producing E. coli (STEC) typically harbor one or both genes that encode for Shiga Toxins I and 2. Nucleic acids are isolated and purified from preserved stool specimens obtained from symptomatic patients exhibiting signs and symptoms of gastroenteritis. This test is intended for use, in conjunction with clinical presentation and epidemiological risk factors, as an aid in the differential diagnosis of Salmonella, Shigella, Campylobacter jejuni/Campylobacter coli, and STEC infections in humans. The results of this test should not be used as the sole basis for diagnosis, treatment, or other patient management decisions. Positive results do not rule out co-infection with other organisms that are are not detected by this test, and may not be the sole or definitive cause of patient illness. Negative ProGastro SSCS Assay results in the setting of clinical illness compatible with gastroenteritis may be due to infection by pathogens that are not detected by this test or non-infectious causes such as ulcerative colitis, irritable bowel syndrome, or Crohn's disease. #### PRODUCT DESCRIPTION The ProGastro SSCS Assay enables detection and differentiation of Salmonella, Shigella, Campylobacter (C. jejuni and C. coli only, undifferentiated) and an Internal Control in the SSC Mix and Shiga Toxin Producing E. coli (STEC, stx1 and stx2 differentiated) and an Internal Control in the STEC Mix. An overview of the procedure is as follows: - 1. Collect raw stool specimens from symptomatic patients and place into Cary Blair Transport Medium or ParaPak C&S (C&S) Transport Medium . - 2. Add the Gastro RNA/DNA Internal Control (GIC) to every sample to monitor for inhibitors present in the specimens. - 3. Perform isolation and purification of nucleic acids using a NucliSENS easyMAG System and the Gen-Probe Prodesse, Inc. JAN 1 6 2013 {1}------------------------------------------------ Automated Magnetic Extraction Reagents (bioMérieux). - 4. Add purified nucleic acids to the SSC Mix included in the ProGastro SSCS Assay Kit. The SSC Mix contains target-specific oligonucleotide primers and probes for detection of Salmonella, Shigella, and Campylobacter (C. jejuni and C. coli only). The primers and probes are complementary to highly conserved regions of genetic sequences for these organisms. The probes are dual-labeled with a reporter dye and a quencher (see table below). - 5. Add purified nucleic acids to the STEC Mix included in the ProGastro SSCS Assay Kit. The STEC Mix contains target-specific oligonucleotide primers and probes for detection of Shiga Toxin 1 and 2 genes (stxl and stx2). The primers and probes are complementary to highly conserved regions of these genes. The probes are dual-labeled with a reporter dye and a quencher (see table below). - 6. Perform amplification of DNA in a Cepheid SmartCycler II instrument. In this process, the probe anneals specifically to the template followed by primer extension and amplification. The ProGastro SSCS Assay is based on Tagman reagent chemistry, which utilizes the 5' - 3' exonuclease activity of Taq polymerase to cleave the probe thus separating the reporter dye from the quencher. This generates an increase in fluorescent signal upon excitation from a light source. With each cycle, additional reporter dye molecules are cleaved from their respective probes, further increasing fluorescent signal. The amount of fluorescence at any given cycle is dependent on the amount of amplification products present at that time. Fluorescent intensity is monitored during each PCR cycle by the real-time instrument. | Supermix | Analyte | Gene Targeted | Probe Fluorophore | Absorbance Peak | Emission Peak | Instrument Channel | |----------------------------|--------------------------------------------------|--------------------------------------|-------------------------|-----------------|---------------|--------------------| | SSC Mix | Campylobacter<br>(C. jejuni and C.<br>coli only) | C. jejuni<br>glyA<br>C. coli<br>cadF | FAM | 495 nm | 520 nm | FAM | | SSC Mix | Salmonella spp. | orgC | CAL Fluor<br>Orange 560 | 538 nm | 559 nm | TET | | SSC Mix | Shigella spp. | ipaH | CAL Fluor<br>Red 610 | 590 nm | 610 nm | Texas Red | | STEC Mix | Shiga Toxin 1 | stx1 | CAL Fluor<br>Orange 560 | 538 nm | 559 nm | TET | | STEC Mix | Shiga Toxin 2 | stx2 | FAM | 495 nm | 520 nm | FAM | | SSC Mix<br>and STEC<br>Mix | Internal Control | NA | Quasar 670 | 647 nm | 670 nm | Cy5 | {2}------------------------------------------------ ## SUBSTANTIAL EQUIVALENCE | Similarities | | | |---------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------| | Element | Prodesse ProGastro SSCS (k123274) | Luminex xTAG GPP (k121454) | | Organisms<br>Detected | Salmonella spp., Shigella spp.,<br>Campylobacter ( <i>C. jejuni</i> and <i>C. coli</i> ),<br>and STEC ( <i>stx1</i> and <i>stx2</i> genes) | Same<br>(See below for differences) | | Analyte | DNA | Same<br>(See below for differences) | | Technological<br>Principles | Multiplex nucleic acid | Same<br>(See below for differences) | | Specimen<br>Types | Stool specimens | Same | | User Complexity | High | Same | | Sample<br>Preparation<br>Method | Up front sample processing is required<br>to extract nucleic acids | Same | | Controls | Internal control in each sample.<br>External control processed with each<br>batch of samples. | Same | | Differences | | | | Element | Prodesse ProGastro SSCS (k123274) | Luminex xTAG GPP (k121454) | | Organisms<br>Detected | (See above for similarities) | Can also detect and distinguish <i>C. lari</i> .<br>In addition, can detect and distinguish<br><i>Clostridium difficile</i> toxin A/B,<br><i>Cryptosporidium</i> ( <i>C. parvum</i> and <i>C.</i><br><i>hominis</i> only), <i>Escherichia coli</i> ( <i>E.</i><br><i>coli</i> ) O157, Enterotoxigenic <i>E. coli</i><br>(ETEC) LT/ST, <i>Giardia</i> ( <i>G. lamblia</i><br>only), Norovirus GI/GII, and<br>Rotavirus A. | | Analyte | DNA | RNA/DNA | | Technological<br>Principles | Real time multiplex PCR based on the<br>Taqman reagent chemistry | Multiplex RT-PCR and bead<br>hybridization followed by<br>Fluorescence-activated sorting of<br>labeled beads coupled to streptavidin-<br>conjugated biotinylated products | | Instrumentation | Cepheid SmartCycler II | PCR Thermocycler and Luminex<br>100/200 system | {3}------------------------------------------------ ### Clinical Performance ### Prospective Study The clinical performance of the ProGastro SSCS Assay was established during prospective studies at four U.S. clinical laboratories. Leftover stool samples were collected during July 2011 - November 2011 and May 2012 - July 2012 and tested during November 2011 thru August 2012. All specimens used in the study meeting the inclusion criteria represented excess remnants of stool specimens that were prospectively collected from symptomatic individuals suspected of gastrointestimal infection, and were submitted for routine care or analysis by each site, and that otherwise would have been discarded. Demographic details for the patient population included in the prospective study are summarized in the following table. | Sex | Number of Samples SSC Mix | Number of Samples STEC Mix | |---------------|---------------------------|----------------------------| | Male | 615/1214 (50.6%) | 615/1214 (50.6%) | | Female | 581/1214 (47.9%) | 581/1214 (47.9%) | | Unknown | 18/1214 (1.5%) | 18/1214 (1.5%) | | Age (yrs) | | | | ≤ 5 years | 378/1214 (31.1%) | 378/1214 (31.1%) | | 6 - 18 years | 296/1214 (24.4%) | 296/1214 (24.4%) | | 19 - 64 years | 357/1214 (29.4%) | 357/1214 (29.4%) | | ≥ 65 years | 164/1214 (13.5%) | 164/1214 (13.5%) | | Unknown | 19/1214 (1.6%) | 19/1214 (1.6%) | Performance of the ProGastro SSCS Assay was assessed and compared to the reference method of culture (Campylobacter, Salmonella, and Shigella) or broth enrichment followed by FDA cleared EIA test (Shiga Toxin producing E. coli). Samples positive for STEC by broth/EIA and/or the ProGastro SSCS Assay underwent PCR followed by bi-directional sequencing to confirm the presence of the stx 1 and/or stx2 genes. Two PCR/sequencing assays were used that each targeted different regions of the stxl or stx2 gene than the ProGastro SSCS Assay. "True" STEC positives were considered as any sample that tested positive for STEC by the broth/ElA method, and "True" STEC negatives were considered as any sample that tested negative for STEC by the broth/EIA method. "True" stx2 positives were considered as any sample that tested positive for STEC by the broth/E/A method and by PCR/sequencing. Bi-directional sequencing data was required to meet pre-defined quality acceptance criteria for both the forward and the reverse sequences that matched stx1 or stx2 sequences deposited in the National Center for Biotechnology Information (NCBI) GenBank database (www.ncbi.nlm.nih.gov), respectively, with acceptable E-values. The E-Value from NCBI BLAST Alignment indicates the statistical significance of a given pair-wise alignment and reflects the size of the database and the scoring system used. The lower the E-Value, the more significant the hit is. A sequence alignment that has an E-Value of le-3 means that this similarity has a 1 in 1000 chance of occurring by chance alone. (http://www.ncbi.nlm.nih.gov/books/bv.fcgi?rid=handbook.section.614). Discrepant results between the ProGastro SSCS Assav and the reference methods were also evaluated using analytically validated PCR/sequencing assays and results are footnoted in the performance tables below. {4}------------------------------------------------ A total of 1214 patients were initially enrolled in the prospective clinical trial. Prospective stool specimens were initially included in the prospective clinical trial. Sixty-one (61) patient/specimens were excluded from the performance calculations due to deviations from the clinical study protocol. Fourteen (14) specimens were excluded for the SSC Mix and 14 were excluded for the STEC Mix from the prospective clinical study data analysis because they remained "Unresolved" after repeat testing with the respective ProGastro SSCS Assay Mix. Unresolved results occur when the sample is negative for all target detections and the Internal Control, indicating potentially PCR-inhibited samples, This resulted in a total of 1139 eligible prospective specimens to be included in the prospective clinical study data analysis. | | <i>Culture</i> | | | | | |-------------------------------------|----------------|----------|-------|------|-----------------------------------------------| | | Positive | Negative | Total | | | | <i>ProGastro<br/>SSCS<br/>Assay</i> | Positive | 20 | 13a | 33 | Sensitivity 100.0%<br>(83.9% - 100.0%) 95% CI | | | Negative | 0 | 1106 | 1106 | Specificity 98.8%<br>(98.0% - 99.3%) 95% CI | | | Total | 20 | 1119 | 1139 | | Campylobacter (C. jejuni / C. coli) Comparison Results a Six (6) samples were positive for Campylobacter (C. coli or C. jejuni) by bi-directional sequence analysis. ### Salmonella Comparison Results | | Culture | | | | |----------------------------|----------|----------|-------|---------------------------------------------| | | Positive | Negative | Total | | | ProGastro<br>SSCS<br>Assay | 20 | 10a | 30 | Sensitivity 95.2%<br>(77.3% - 99.2%) 95% CI | | | 1b | 1108 | 1109 | Specificity 99.1%<br>(98.4% - 99.5%) 95% CI | | Total | 21 | 1118 | 1139 | | a Ten (10) samples were positive for Salmonella by bi-directional sequence analysis. Sample was positive for Salmonella by bi-directional sequence analysis. ### Shigella Comparison Results | | Culture | | | | | |----------------------------|----------|----------|-------|------|-----------------------------------------------| | | Positive | Negative | Total | | | | ProGastro<br>SSCS<br>Assay | Positive | 15 | 6a | 21 | Sensitivity 100.0%<br>(79.6% - 100.0%) 95% CI | | | Negative | 0 | 1118 | 1118 | Specificity 99.5%<br>(98.8% - 99.8%) 95% CI | | | Total | 15 | 1124 | 1139 | | a Six (6) samples were positive for Shigella by bi-directional sequence analysis. {5}------------------------------------------------ ### STEC Comparison Results | | Broth Enrichment/EIA | | | | | |----------------------------|----------------------|----------|-------|-------------|-----------------------------------------------| | | Positive | Negative | Total | | | | ProGastro<br>SSCS<br>Assay | Positive | 9a | 9b | <b>18</b> | Sensitivity 100.0%<br>(70.1% - 100.0%) 95% CI | | | Negative | 0 | 1121 | <b>1121</b> | Specificity 99.2%<br>(98.5% - 99.6%) 95% CI | | | Total | 9 | 1130 | 1139 | | a Six (6) samples positive for stx1, one (1) sample positive for stx2, and two (2) samples positive for stx1 and stx2 by bi-directional sequence analysis. b Six (6) samples positive for stx1 and three (3) samples positive for stx2 by bi-directional sequence analysis. ### stx1 Comparison Results | | Broth Enrichment/EIA and<br>sequencing for stx1 | | | | | |-------------------------|-------------------------------------------------|----------|-------|----|-----------------------------------------------------------------| | | Positive | Negative | Total | | | | ProGastro<br>SSCS Assay | Positive | 8 | 6a | 14 | Positive Percent Agreement<br>100.0%<br>(67.6% - 100.0%) 95% CI | | | Negative | 0 | 4 | 4 | Negative Percent Agreement 40.0%<br>(16.8% - 68.7%) 95% CI | | | Total | 8 | 10 | 18 | | 4 Six (6) samples negative by broth/EIA, five (5) were positive for stx/by bi-directional sequencing, but were negative by Broth Enrichment/EIA. ### stx2 Comparison Results | | Broth Enrichment/EIA<br>and sequencing for stx2 | | | | | |-------------------------|-------------------------------------------------|----------|-------|----|-----------------------------------------------------------------| | | Positive | Negative | Total | | | | ProGastro<br>SSCS Assay | Positive | 3 | 3a | 6 | Positive Percent Agreement<br>100.0%<br>(43.9% - 100.0%) 95% CI | | | Negative | 0 | 12 | 12 | Negative Percent Agreement 80.0%<br>(54.8% - 93.0%) 95% CI | | | Total | 3 | 15 | 18 | | 4 Three (3) samples were positive for stx2 by bi-directional sequence analysis, but were negative by Broth Enrichment/EIA. {6}------------------------------------------------ The ProGastro SSCS Assay detected one mixed infections in the prospective clinical evaluation. This represents 0.98% of the total positive specimens (1/102). The one mixed infections sample was double infections and was confirmed by the reference methods. Distinct Co-infection Combinations Detected by the ProGastro SSCS Assay in the Prospective Clinical Trial | Distinct Co-infection Combinations<br>Detected by ProGastro SSCS Assay | | | | | | |------------------------------------------------------------------------|---------------|-----------|------------------------|-------------------------------------------|------------------------| | Analyte 1 | Analyte 2 | Analyte 3 | Total<br>Co-infections | Number of<br>Discrepant<br>Co-infectionsa | Discrepant Analyte(s)a | | Salmonella | Campylobacter | N/A | 0 | 0 | | | Salmonella | Shigella | N/A | 0 | 0 | | | Salmonella | STEC | N/A | 0 | 0 | | | Campylobacter | Shigella | N/A | 0 | 0 | | | Campylobacter | STEC | N/A | 1 | 0 | | | STEC | Shigella | N/A | 0 | 0 | | | Salmonella | Campylobacter | STEC | 0 | 0 | | | Total Co-infections | | | 1 | 0 | | | Total Double Infections | | | 1 | 0 | | | Total Triple Infections | | | 0 | 0 | | ª A discrepant co-infection or discrepant analyte was defined as one that was detected by the ProGastro SSCS Assay but not detected by the reference methods. There were no co-infections that were detected by the reference method and not detected by the ProGastro SSCS Assay. ### Retrospective Study In addition to the prospective clinical study, two clinical sites also performed testing using retrospective samples that were collected from 2007 - 2011. A total of 105 stool samples were included in the retrospective study. These samples had been previously determined to be positive or negative by culture and/or Broth Enrichment/ElA. The ProGastro SSCS Assay was compared to the same reference method that was employed for the prospective study to determine positive and negative percent agreement. Demographic details for this patient population are summarized in the table below. | Sex* | Number of Subjects | |---------------|--------------------| | Female | 24/55 (43.6%) | | Male | 31/55 (56.4%) | | Age | Number of Subjects | | ≤ 5 years | 12/105 (11.4%) | | 6 - 18 years | 24/105 (22.9%) | | 19 – 64 years | 51/105 (48.6%) | | ≥ 65 years | 18/105 (17.1%) | *For all of the 50 specimens tested from one site the gender was unknown {7}------------------------------------------------ Campylobacter Comparison Results | | Culture | | | | | |-------------------------|----------|----------|----------|-------|---------------------------------------------------------------| | | | Positive | Negative | Total | | | ProGastro<br>SSCS Assay | Positive | 27 | 5 | 32 | Positive Percent Agreement 96.4%<br>(82.3% - 99.4%) 95% CI | | | Negative | 1 | 72 | 73 | Negative Percent Agreement<br>93.5%<br>(85.7% - 97.2%) 95% CI | | | Total | 28 | 77 | 105 | | ### Salmonella Comparison Results | | | | Culture | | | | |-------------------------|----------|---|----------|----------|-------|-----------------------------------------------------------------| | | | | Positive | Negative | Total | | | ProGastro<br>SSCS Assay | Positive | 3 | 0 | 3 | | Positive Percent Agreement<br>100.0%<br>(43.4% - 100.0%) 95% CI | | | Negative | 0 | 102 | 102 | | Negative Percent Agreement<br>100.0%<br>(96.4% - 100.0%) 95% CI | | Total | | 3 | 102 | 105 | | | ### Shigella Comparison Results | | | Culture | | | | |-------------------------|----------|----------|----------|-------|-----------------------------------------------------------------| | | | Positive | Negative | Total | | | ProGastro<br>SSCS Assay | Positive | 4 | 0 | 4 | Positive Percent Agreement<br>100.0%<br>(51.0% - 100.0%) 95% CI | | | Negative | 0 | 101 | 101 | Negative Percent Agreement<br>100.0%<br>(96.3% - 100.0%) 95% CI | | Total | | 4 | 101 | 105 | | ### STEC Comparison Results | | | | Culture or Broth Enrichment/EIA | | | | | |----------------------------|----------|-------|---------------------------------|----------|-------|--------------------------------------------------------------|--| | | | | Positive | Negative | Total | | | | ProGastro<br>SSCS<br>Assay | Positive | | 19a | 0 | 19 | Positive Percent Agreement 100.0%<br>(83.2% - 100.0%) 95% CI | | | | Negative | | 0 | 86 | 86 | Negative Percent Agreement 100.0%<br>(95.7% - 100.0%) 95% CI | | | | | Total | 19 | 86 | 105 | | | 4 Five (5) samples positive for stx1, 5 samples positive for stx2, and 9 samples positive for stx1 and stx2. {8}------------------------------------------------ ### stx1 Comparison Results | | Culture or Broth Enrichment/EIA<br>and sequencing for stx1 | | | | | |----------------------------|------------------------------------------------------------|----------|----------|-------|--------------------------------------------------------------| | ProGastro<br>SSCS<br>Assay | | Positive | Negative | Total | | | | Positive | 14 | 0 | 14 | Positive Percent Agreement 100.0%<br>(78.5% - 100.0%) 95% CI | | | Negative | 0 | 5 | 5 | Negative Percent Agreement 100.0%<br>(56.6% - 100.0%) 95% CI | | | Total | 14 | 5 | 19 | | ### stx2 Comparison Results | | | Culture or Broth Enrichment/EIA<br>and sequencing for stx2 | | | | |----------------------------|----------|------------------------------------------------------------|----------|-------|--------------------------------------------------------------| | | | Positive | Negative | Total | | | ProGastro<br>SSCS<br>Assay | Positive | 14 | 0 | 14 | Positive Percent Agreement 100.0%<br>(78.5% - 100.0%) 95% CI | | | Negative | 0 | 5 | 5 | Negative Percent Agreement 100.0%<br>(56.6% - 100.0%) 95% CI | | | Total | 14 | 5 | 19 | | Of the prospective and retrospective specimens run using the SSC Mix Assay, 98.0% (1233/1258) of these specimens were successful on the first attempt. The remaining 25 (25/1258 = 2.0%) gave "Unresolved" results on the first attempt. An "Unresolved" result is generated when the Gastro Internal Control (GIC) fails to be detected in a clinical specimen. A failure of the GIC to be detected can occur if inhibitors are present in a sample or due to technical error (e.g., GIC not added prior to nucleic acid extraction). Of the 25 "Unresolved" specimens on the first attempt with sufficient nucleic acid for retest, 44.0% (11/25) gave a valid result on the second attempt. The remaining 14 were "Unresolved" on the second attempt. Of the prospective and retrospective specimens run using the STEC Mix Assay, 97.9% (1232/1258) of these specimens were successful on the first attempt. The remaining 26 (26/1258 = 2.1%) gave "Unresolved" results on the first attempt. Of the 26 "Unresolved" specimens on the first attempt with sufficient nucleic acid for retest, 48.0% (12/25) gave a valid result on the second attempt. The remaining 14 were "Unresolved" on the second attempt. #### Reproducibility The reproducibility of the ProGastro SSCS Assay was evaluated at three laboratory sites. Reproducibility was assessed using a panel of 15 simulated samples that included medium positive, low positive (near the assav limit of detection, ≥ 95% positive), and high negative (below the assay limit of detection, ≤ 95% positive) samples for each of the assay targets. The reproducibility panel and controls were run with the ProGastro SSCS Assay (SSC and STEC Mixes) at three sites by each of two operators per site for five days. {9}------------------------------------------------ ### Page 10 of 11 01/15/2013 #### Reproducibility Panel Member Results | | SSC Mix | | | | | | | | STEC Mix | | | | | | SSC STEC<br>Mix Mix | | |--------|-----------------------------------------------|-------------------------------|----------------------------------|-----------------------------|--------------------------------|--------------------------------|--------------------------------------|------------------------------|---------------------------------|---------------------------|---------------------------------|---------------------------|---------------------------------|--------------------------------|--------------------------------|--| | | Panel<br>Member ID | <i>C. jejuni</i> Low Positive | <i>C. jejuni</i> Medium Positive | <i>C. coli</i> Low Positive | <i>C. coli</i> Medium Positive | <i>Salmonella</i> Low Positive | <i>Salmonella</i> Medium<br>Positive | <i>Shigella</i> Low Positive | <i>Shigella</i> Medium Positive | STEC (stx 1) Low Positive | STEC (stx 1) Medium<br>Positive | STEC (stx 2) Low Positive | STEC (stx 2) Medium<br>Positive | High Negative (IC Ct<br>Value) | High Negative (IC Ct<br>Value) | | | | Concentration | 20X*<br>LoD | 100X*<br>LoD | 6X*<br>LoD | 30X*<br>LoD | 2X<br>LoD | 10X<br>LoD | 2X<br>LoD | 10X<br>LoD | 2X<br>LoD | 10X<br>LoD | 2X<br>LoD | 10X<br>LoD | 0.0001X<br>LoD | | | | | Agreement<br>with Expected<br>Result | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 90/90<br>100% | 90/90<br>100% | | | Site 1 | Mean Ct Value | 37.6 | 35.2 | 35.9 | 33.5 | 35.9 | 33.2 | 35.8 | 33.2 | 36.1 | 33.4 | 36.7 | 34.5 | 33.6 | 33.2 | | | | % CV | 3.5 | 3.0 | 3.9 | 3.7 | 1.4 | 1.2 | 1.7 | 1.4 | 2.0 | 1.6 | 1.8 | 1.3 | 3.0 | 1.7 | | | | Agreement<br>with Expected<br>Result | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 29/30<br>96.7% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 90/90<br>100% | 90/90<br>100% | | | Site 2 | Mean Ct Value | 37.3 | 34.8 | 35.8 | 33.4 | 36.0 | 33.1 | 35.6 | 33.2 | 36.2 | 33.6 | 36.8 | 34.7 | 33.3 | 33.0 | | | | % CV | 3.0 | 2.8 | 3.1 | 3.2 | 1.7 | 1.6 | 1.9 | 1.8 | 2.5 | 1.6 | 1.9 | 1.9 | 0.9 | 0.9 | | | | Agreement<br>with Expected<br>Result | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 30/30<br>100% | 90/90<br>100% | 90/90<br>100% | | | Site 3 | Mean Ct Value | 37.0 | 34.4 | 35.2 | 32.9 | 35.7 | 32.7 | 35.0 | 32.8 | 35.6 | 33.1 | 36.5 | 34.2 | 32.9 | 32.6 | | | | % CV | 2.6 | 2.2 | 2.1 | 1.9 | 1.4 | 1.2 | 1.8 | 1.3 | 1.7 | 1.3 | 1.6 | 1.0 | 0.8 | 1.1 | | | | Total<br>Agreement<br>with Expected<br>Result | 90/90<br>100% | 90/90<br>100% | 90/90<br>100% | 90/90<br>100% | 90/90<br>100% | 90/90<br>100% | 89/90<br>89.9% | 90/90<br>100% | 90/90<br>100% | 90/90<br>100% | 90/90<br>100% | 90/90<br>100% | 270/270<br>100% | 270/270<br>100% | | | | 95% CI | 95.9%-<br>100.0% | 95.9%-<br>100.0% | 95.9%-<br>100.0% | 95.9%-<br>100.0% | 95.9%-<br>100.0% | 95.9%-<br>100.0% | 94.0%-<br>99.8% | 95.9%-<br>100.0% | 95.9%-<br>100.0% | 95.9%-<br>100.0% | 95.9%-<br>100.0% | 95.9%-<br>100.0% | 98.6%-<br>100.0% | 98.6%-<br>100.0% | | | | Overall Mean<br>Ct Value | 37.3 | 34.8 | 35.6 | 33.3 | 35.9 | 33.0 | 35.5 | 33.1 | 35.9 | 33.4 | 36.7 | 34.5 | 33.2 | 32.9 | | | | Overall<br>% CV | 3.1 | 2.8 | 3.2 | 3.1 | 1.6 | 1.5 | 2.0 | 1.6 | 2.2 | 1.6 | 1.8 | 1.5 | 2.1 | 1.5 | | * Note: The Campylobacter strains were tested at higher concentrations because Campylobacter is sensitive to environmental stressors including freezing where it loses viability. However, the average Ct value for the C. jejuni (ATCC 33291) Low Positives was 37.3, which is very close to the average Ct value of 38.8 for the same C. jejuni strain tested at the estimated LoD level in the Analytical Reactivity Study. The average Ct value for the C. coli (ATCC BAA-371) Low Positives was 35.6, which is very close to the average Ct value of 34.4 for the same C. coli strain tested at the estimated LoD level in the Analytical Reactivity Study. Therefore, the effective DNA concentrations of the C. jejuni (ATCC 33291) and C. codi (ATCC BAA-371) low positive samples tested in the Reproducibility Study are very close to the estimated LoD DNA concentrations for these two strains tested in the Analytical Reactivity Study. {10}------------------------------------------------ #### Precision The precision of the ProGastro SSCS Assay was evaluated internally using a panel of 15 simulated samples that included medium positive (near the assay limit of detection, ≥ 95% positive), and high negative (below the assay limit of detection, ≤ 95% positive) samples for each of the assay targets. A panel of 15 contrived samples including the necessary controls was run with the ProGastro SSCS Assay (SSC and STEC Mixes) by each of two operators for twelve days. | | SSC Mix | | | | | | | | STEC Mix | | | | SSC<br>Mix | STEC<br>Mix | |-----------------------------------------------|-------------------------------|----------------------------------|-----------------------------|--------------------------------|--------------------------------|-----------------------------------|------------------------------|---------------------------------|------------------------------------|---------------------------------------|------------------------------------|---------------------------------------|-----------------------------|-----------------------------| | Panel<br>Member ID | <i>C. jejuni</i> Low Positive | <i>C. jejuni</i> Medium Positive | <i>C. coli</i> Low Positive | <i>C. coli</i> Medium Positive | <i>Salmonella</i> Low Positive | <i>Salmonella</i> Medium Positive | <i>Shigella</i> Low Positive | <i>Shigella</i> Medium Positive | STEC ( <i>stx 1</i> ) Low Positive | STEC ( <i>stx 1</i> ) Medium Positive | STEC ( <i>stx 2</i> ) Low Positive | STEC ( <i>stx 2</i> ) Medium Positive | High Negative (IC Ct Value) | High Negative (IC Ct Value) | | Concentration | 20X*<br>LoD | 100X*<br>LoD | 6X*<br>LoD | 30X*<br>LoD | 2X<br>LoD | 10X<br>LoD | 2X<br>LoD | 10X<br>LoD | 2X<br>LoD | 10X<br>LoD | 2X<br>LoD | 10X<br>LoD | 0.0001X LoD | 0.0001X LoD | | Total<br>Agreement<br>with Expected<br>Result | 72/72<br>100% | 72/72<br>100% | 72/72<br>100% | 72/72<br>100% | 72/72<br>100% | 72/72<br>100% | 72/72<br>100% | 72/72<br>100% | 72/72<br>100% | 72/72<br>100% | 72/72<br>100% | 72/72<br>100% | 71/72<br>98.6% | 72/72<br>100% | | Overall Mean<br>Ct Value | 36.7 | 34.2 | 35.0 | 32.7 | 35.6 | 32.7 | 35.1 | 32.7 | 35.6 | 33.0 | 36.5 | 34.2 | 32.9 | 32.5 | | Overall % CV | 2.6 | 2.4 | 2.1 | 2.3 | 1.5 | 1.1 | 1.6 | 1.3 | 1.8 | 1.3 | 1.8 | 1.2 | 0.9 | 1.0 | ### Precision Panel Member Results Note: The Campylobacter strains were tested at higher concentrations because Campylobacter is sensitive to environmental stressors including freezing where it loses viability. However, the average Ct value for the C. jejuni (ATCC 33291) Low Positives was 37.3, which is very close to the average Ct value of 38.8 for the same C. jejuni strain tested at the estimated LoD level in the Analytical Reactivity Study. The average Ct value for the C. coli (ATCC BAA-371) Low Positives was 35.6, which is very close to the average Ct value of 34.4 for the same C. coli strain tested at the estimated LoD level in the Analytical Reactivity Study. Therefore, the effective DNA concentrations of the C. jejuni (ATCC 33291) and C. codi (ATCC BAA-371) low positive samples tested in the Precision Study are very close to the estimated LoD DNA concentrations for these two strains tested in the Analytical Reactivity Study. {11}------------------------------------------------ Image /page/11/Picture/0 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter. Inside the circle is a stylized symbol resembling an abstract human figure with three flowing lines extending from its head. #### DEPARTMENT OF HEALTH & HUMAN SERVICES #### Public Health Service Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-002 Gen-Probe Prodesse, Inc. c/o Karen Harrington, Ph.D. Manager, Clinical Affairs 20925 Crossroads Circle Waukesha, WI 53186 JAN 1 6 2013 Re: k123274 Trade/Device Name: ProGastro SSCS Assay Regulation Number: 21 CFR 866.3990 Regulation Name: Gastrointestinal Microorganism Multiplex Nucleic Acid-Based Assay Regulatory Class: Class II Product Code: PCH, PCI, OOI Dated: October 17. 2012 Received: October 19, 2012 Dear Dr. Harrington: We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register. Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set {12}------------------------------------------------ Page 2 -- Karen Harrington forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050. If you desire specific advice for your device on our labeling regulation (21 CFR Parts 801 and 809), please contact the Office of In Vitro Diagnostics and Radiological Health at (301) 796-5450. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance. You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html. Sincerely yours, Uwe Scherf for Sally A. Hojvat, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health Enclosure {13}------------------------------------------------ # Indication for Use 510(k) Number: k123274 Device Name: ProGastro SSCS Assay Indication For Use: The Prodesse® ProGastro SSCS Assay is a multiplex real time PCR in vitro diagnostic test for the qualitative detection and differentiation of Salmonella, Shigella, and Campylobacter (C. jejuni and C. coli only, undifferentiated) nucleic acids and Shiga Toxin 1 (stxl) and Shiga Toxin 2 (stx2) genes. Shiga toxin producing E. coli (STEC) typically harbor one or both genes that encode for Shiga Toxins 1 and 2. Nucleic acids are isolated and purified from preserved stool specimens obtained from symptomatic patients exhibiting signs and symptoms of gastroenteritis. This test is intended for use, in conjunction with clinical presentation and epidemiological risk factors, as an aid in the differential diagnosis of Salmonella, Shigella, Campylobacter jejuni/Campylobacter coli, and STEC infections in humans. The results of this test should not be used as the sole basis for diagnosis, treatment, or other patient management decisions. Positive results do not rule out co-infection with other organisms that are not detected by this test, and may not be the sole or definitive cause of patient illness. Negative ProGastro SSCS Assay results in the setting of clinical illness compatible with gastroenteritis may be due to infection by pathogens that are not detected by this test or non-infectious causes such as ulcerative colitis, irritable bowel syndrome, or Crohn's disease. Prescription Use X (21 CFR Part 801 Subpart D) And/Or Over the Counter Use _ (21 CFR Part 801 Subpart C) (PLEASE DO NOT WRITE BELOW THIS LINE; CONTINUE ON ANOTHER PAGE IF NEEDED) Concurrence of CDRH, Office of In Vitro Diagnostics and Radiological Health (OIR) R. H. Hitz ion Sign-Off Office of In Vitro Diagnostics and Radiological Health k 23274