XTAG GASTROINTESTINAL PATHOGEN PANEL(GPP)/XTAG DATA ANALYSIS SOFTWARE FOR GPP(TDAS GPP)

{0}------------------------------------------------ Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health and Human Services. The logo features the department's name encircling a symbol. The symbol consists of three stylized human profiles facing right, layered on top of each other. The profiles are connected by a flowing, ribbon-like shape. Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002 October 24, 2014 LUMINEX MOLECULAR DIAGNOSTICS, INC. TINA IP REGULATORY AFFAIRS ASSOCIATE 439 UNIVERSITY AVE. TORONTO, ONTARIO, M5G 1Y8 CANADA Re: K140647 Trade/Device Name: Xtag® Gastrointestinal Pathogen Panel (GPP) Regulation Number: 21 CFR 866.3990 Regulation Name: Gastrointestinal pathogen panel multiplex nucleic acid-based assay system Regulatory Class: II Product Code: PCH, NSU, JJH Dated: September 15, 2014 Received: September 16, 2014 Dear Ms. Ip: 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. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading. If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register. Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal 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 Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the {1}------------------------------------------------ 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 regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. 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 Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. Sincerely yours, # Stephen J. Lovell -S 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 {2}------------------------------------------------ # Indications for Use #### 510(k) Number (if known) K140647 #### Device Name xTAG® Gastrointestinal Pathogen Panel (GPP) #### Indications for Use (Describe) The xTAG® Gastrointestinal Pathogen Panel (GPP) is a multiplexed nucles squalitative detection and identification of multiple viral, bacterial and parasitic nucleic acids in human stool in Cary-Blair media from individuals with signs and symptoms of infectious of infectious of infectious of infectio or gastroenteritis. The following pathogen types, subtypes and toxin genes are identified using the xTAG GPP. #### Viruses - · Adenovirus 40/41 - · Norovirus GI/GII - Rotavirus A #### Bacteria - · Campylobacter (C. jejuni, C. coli and C. lari only) - · Clostridium difficile (C. difficile) toxin A/B - · Escherichia coli (E. coli) O157 - · Enterotoxigenic E. coli (ETEC) LT/ST - · Salmonella - · Shiga-like Toxin producing E. coli (STEC) stx 1/stx 2 - · Shigella (S. boydii, S. sonnei, S. flexneri and S. dysenteriae) - Vibrio cholerae (V. cholerae) cholera toxin gene (ctx) #### Parasites - · Cryptosporidium (C. parvum and C. hominis only) - Entamoeba histolytica (E. histolytica) - · Giardia (G. lamblia only, also known as G. intestinalis and G. duodenalis) The detection and identification of specific and microbial nucleic acid from individuals exhibiting signs and symptoms of gastrontestinal infection ads in the digencis of gastrontestinal infection when with clinical evaluation, laboratory findings and epidemological information. A gastroinestinal microorganism multiplex nucleic acid-based assay also in the detection and identification of acute gastroenteritis in the context of outbreaks. #### xTAG GPP positive results are presumptive and must be confirmed by FDA-cleared tests or other acceptable reference methods. The results of this test should not be used as for diagnosis, treatment, or other patient management decisions. Confirmed positive results do not rule out co-infection with other organisms that are not dethe sole or definitive cause of patient illness. Negative xTAG GPP results in the setting of clinical illness compatible with gastroently pathogens that are not detected by this test or non-infectious causes such as ulcerative colitis, irritable bowel syndrome, or Crohn's disease. #### xTAG GPP is not intended to monitor or guide treatment for C. difficile infections. The xTAG GPP test is indicated for use with the Lumines® 100/200™ and MAGPIX® instruments with xPONENT® software. #### Type of Use (Select one or both, as applicable) X | Prescription Use (Part 21 CFR 801 Subpart D) Over-The-Counter Use (21 CFR 801 Subpart C) #### PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON A SEPARATE PAGE IF NEEDED. #### FOR FDA USE ONLY Concurrence of Center for Devices and Radiological Health (CDRH) (Signature) {3}------------------------------------------------ This section applies only to requirements of the Paperwork Reduction Act of 1995. #### *DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.* The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to: > Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff(@fda.hhs.gov "An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number." {4}------------------------------------------------ Image /page/4/Picture/0 description: The image shows the word "Luminex" in a bold, sans-serif font. The word is black, except for a red dot above the "i". The word is slightly slanted to the right. A small registered trademark symbol is to the right of the "x". # 510(k) Summary This Summary of 510(k) information is being submitted in accordance with the requirements of 21 CFR 807.92. #### 510(k) Number: K140647 Submission Type: Traditional 510(k), New Device Measurand: A panel of viruses, bacteria and parasites and toxins including: Adenovirus 40/41, Norovirus GI/GII, Rotavirus A, Campylobacter (C. jejuni, C. coli and C. lari only), Clostridium difficile toxin A/B, Escherichia coli (E. coli) O157, Enterotoxigenic E. coli (ETEC) LT/ST, Salmonella, Shiga-like Toxin producing E. coli (STEC) stx 1/stx 2, Shigella (S. boydii, S. sonnei, S. flexneri and S. dysenteriae), Vibrio cholerae (V. cholera toxin gene (ctx), Cryptosporidium (C. parvum and C. hominis only), Entamoeba histolytica (E. histolytica), Giardia (G. lamblia only, also known as G. intestinalis and G. duodenalis) and the internal control (bacteriophage MS2). Type of Test: Qualitative nucleic acid multiplex test Applicant: Luminex Molecular Diagnostics, Inc., Toronto, Ontario, Canada Proprietary and Established Names: xTAG Gastrointestinal Pathogen Panel (GPP) #### Regulatory Information: | Product Code | Classification | Regulation Section | Review Panel | |--------------|----------------|--------------------------------------------------------------------------------------------|-------------------| | PCH | II | 21CFR866.3990 Gastrointestinal Pathogen Panel<br>Multiplex Nucleic Acid-Based Assay System | Microbiology (83) | | NSU | II | 21CFR862.2570 Multiplex Instrument System | Microbiology (83) | #### Device Components | Product | Description | |------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------| | xTAG® GPP Kit | Unchanged from k121894 | | xTAG® GPP TDAS (Software CD) | Revised CD, containing data acquisition protocol and data analysis<br>software (updated to include Adenovirus 40/41, V. cholerae and E.<br>histolytica) | | Luminex® MAGPIX® instrument | Unchanged from k121894 | | xPONENT® Software | xPONENT® Software unchanged from k121894 | {5}------------------------------------------------ Image /page/5/Picture/0 description: The image shows the logo for Luminex. The logo is black, with a red dot above the "i" in Luminex. The font is bold and sans-serif. #### Intended Use: The xTAG® Gastrointestinal Pathogen Panel (GPP) is a multiplexed nucleic acid test intended for the simultaneous qualitative detection and identification of multiple viral, bacterial, and parasitic nucleic acids in human stool specimens or human stool in Cary-Blair media from individuals with signs and symptoms of infectious colitis or gastroenteritis. The following pathogen types, subtypes and toxin genes are identified using the xTAG GPP: #### Viruses - . Adenovirus 40/41 - Norovirus GI/GII ● - Rotavirus A #### Bacteria - Campylobacter (C. jejuni, C. coli and C. lari only) ● - Clostridium difficile (C. difficile) toxin A/B - Escherichia coli (E. coli) 0157 - Enterotoxigenic Escherichia coli (ETEC) LT/ST - . Salmonella - Shiga-like Toxin producing E. coli (STEC) stx 1/stx 2 - Shigella (S. boydii, S. sonnei, S. flexneri and S. dysenteriae) - Vibrio cholerae (V. cholerae) cholera toxin gene (ctx) ● #### Parasites - Cryptosporidium (C. parvum and C. hominis only) ● - Entamoeba histolytica (E. histolytica) - . Giardia (G. lamblia only, also known as G. intestinalis and G. duodenalis) The detection and identification of specific gastrointestinal microbial nucleic acid from individuals exhibiting signs and symptoms of gastrointestinal infection aids in the diagnosis of gastrointestinal infection when used in conjunction with clinical evaluation, laboratory findings and epidemiological information. A gastrointestinal microorganism multiplex nucleic acid-based assay also aids in the detection and identification of acute gastroenteritis in the context of outbreaks. ## xTAG GPP positive results are presumptive and must be confirmed by FDA-cleared tests or other acceptable reference methods. The results of this test should not be used as the sole basis for diagnosis, treatment, or other patient management decisions. Confirmed 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 xTAG GPP 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 noninfectious causes such as ulcerative colitis, irritable bowel syndrome, or Crohn's disease. #### xTAG GPP is not intended to monitor or guide treatment for C. difficile infections. {6}------------------------------------------------ Image /page/6/Picture/0 description: The image shows the word "Luminex" in a bold, sans-serif font. The word is black, except for a red dot above the "i". The dot is slightly offset to the right. The xTAG GPP test is indicated for use with the Luminex® 100/200™ and MAGPIX® instruments with xPONENT® software. Indication(s) for use: Same as intended use. Special instrument requirements: Luminex MAGPIX instrument with xPONENT software. #### Device Description: xTAG GPP incorporates a multiplex reverse-transcription polymerase chain reaction (RT-PCR) with Luminex's proprietary universal sorting system (the xTAG Universal Array) on the Luminex platform. The xTAG Universal Array sorts nucleic acids onto discreet Luminex bead populations by virtue of highly specific "tag/anti-tag" hybridization reactions. The tags and anti-tags comprising the xTAG Universal Array are 24-mer oligonucleotide sequences not found in nature. The assay has been designed to simultaneously detect microbial targets and an internal control (bacteriophage MS2 added to each sample prior to extraction). For each sample, 10 µL of extracted nucleic acid is amplified in a single multiplex RT-PCR reaction. Amplimers ranging from 58 to 202 bp (not including the 24-mer tag) are generated in this reaction. A five μL aliquot of the RT-PCR product is then subjected to a hybridization/detection reaction that also includes bead populations coupled to 24-mer antitags. Each bead population is coupled to a unique anti-tag which is the exact complement of a 24-mer tag incorporated into a given amplimer. Thus, each Luminex bead population uniquely identifies a microbial target or assay control through a specific tag/anti-tag hybridization reaction. Signal is generated via a Streptavidin, R-Phycoerythrin conjugate. The Luminex instrument sorts the products of these hybridization reactions and generates a signal in the form of a median fluorescence intensity (MFI) value for each bead population. The MFI values are generated by the xPONENT software provided with the instrument using the GPP protocol parameters, and are analyzed by the xTAG Data Analysis Software (TDAS GPP (US)). TDAS GPP (US) applies algorithms to MFI values in order to generate a qualitative result for each microbial target selected for reporting to establish the presence or absence of bacterial, viral or parasitic targets and/or controls in each sample. The data analysis software also generates a qualitative result and compiles a report for patient samples and external controls assayed in a given run. Before data are analyzed, a user has the option to select a subset of the targets from the intended use of the xTAG GPP (for each sample). {7}------------------------------------------------ Image /page/7/Picture/0 description: The image shows the word "Luminex" in a bold, sans-serif font. The word is black, except for a red dot above the "i". A small registration mark is to the right of the "x". #### Substantial Equivalence Information: | Item | New Device (k140647) xTAG GPP | Predicate (k121894) xTAG GPP | |------------------------------------------------------|------------------------------------------|-----------------------------------------| | Manufacturer (Same) | Luminex Molecular Diagnostics | Luminex Molecular Diagnostics | | Extraction Method (Same) | bioMérieux NucliSENS® easyMAG® | bioMérieux NucliSENS easyMAG | | Test Principle and<br>Amplification Method<br>(Same) | Multiplex end point RT-PCR | Multiplex end point RT-PCR | | | | | | | | | | Kit Reagents (Same) | xTAG GPP Primer Mix, xTAG OneStep | xTAG GPP Primer Mix, xTAG OneStep | | | Enzyme Mix, xTAG® OneStep Buffer, | Enzyme Mix, xTAG OneStep Buffer, | | | xTAG RNase-Free Water, xTAG BSA, | xTAG RNase-Free Water, xTAG BSA, | | | xTAG MS2, xTAG® GPP Bead Mix, xTAG | xTAG MS2, xTAG GPP Bead Mix, xTAG | | | Reporter Buffer, xTAG 0.22 SAPE | Reporter Buffer, xTAG 0.22 SAPE | | Test Format (Same) | Multiplex MAGPLEX bead-based | Multiplex MAGPLEX bead-based | | | universal array | universal array | | Detection Method (Same) | Fluorescence based | Fluorescence based | | Quality Control (Same) | Internal Control (MS2), rotating analyte | Internal Control (MS2), rotating | | | controls and negative control (RNAse- | analyte controls and negative control | | | free water) | (RNAse-free water) | | Results (Same) | Qualitative | Qualitative | | Instrument Software<br>System | Luminex MAGPIX with xPONENT<br>Software | Luminex MAGPIX with xPONENT<br>Software | Table 1: Similarities between New Device and Predicate ### Table 2: Differences between New Device and Predicate | Item | New Device (k140647)<br>xTAG GPP | Predicate (k121894)<br>xTAG GPP | |---------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------| | Specimen<br>Types | Human stool specimens and human stool in<br>Cary-Blair media | Human stool specimens | | Software | Updated assay protocol to acquire and show<br>data for additional 3 analytes: Adenovirus<br>40/41, Entamoeba histolytica (E. histolytica),<br>and Vibrio cholerae (V. cholerae). | Assay protocol file excludes analytes<br>Adenovirus 40/41, Entamoeba histolytica (E.<br>histolytica), and Vibrio cholerae (V. cholerae) | | Intended<br>Use | See above. Addition of sample type human<br>stool in Cary-Blair media and addition of<br>analytes Adenovirus 40/41, Entamoeba<br>histolytica (E. histolytica), and Vibrio cholerae<br>(V. cholerae) cholera toxin gene (ctx).<br>Specified software used with Luminex 100/200<br>(k140377) and MAGPIX instruments.<br>Organized analytes listed under sub-heading of<br>viruses, bacteria and parasites. | The xTAG Gastrointestinal Pathogen Panel<br>(GPP) is a multiplexed nucleic acid test intended<br>for the simultaneous qualitative detection and<br>identification of multiple viral, parasitic, and<br>bacterial nucleic acids in human stool<br>specimens from individuals with signs and<br>symptoms of infectious colitis or gastroenteritis.<br>The following pathogen types, subtypes and<br>toxin genes are identified using the xTAG® GPP:<br>• Campylobacter (C. jejuni, C. coli and C. lari<br>only)<br>• Clostridium difficile (C. difficile) toxin A/B<br>• Cryptosporidium (C. parvum and C. hominis<br>only)<br>• Escherichia coli (E. coli) O157<br>• Enterotoxigenic Escherichia coli (ETEC) LT/ST<br>• Giardia (G. lamblia only - also known as G.<br>lamblia and G. duodenalis) | | Item | New Device (k140647) | Predicate (k121894) | | | xTAG GPP | xTAG GPP | | | | • Norovirus GI/GII | | | | • Rotavirus A | | | | • Salmonella | | | | • Shiga-like Toxin producing E. coli (STEC) stx 1/stx 2 | | | | • Shigella (S. boydii, S. sonnei, S. flexneri and S. dysenteriae) | | | | The detection and identification of specific gastrointestinal microbial nucleic acid from individuals exhibiting signs and symptoms of gastrointestinal infection aids in the diagnosis of gastrointestinal infection when used in conjunction with clinical evaluation, laboratory findings and epidemiological information. A gastrointestinal microorganism multiplex nucleic acid-based assay also aids in the detection and identification of acute gastroenteritis in the context of outbreaks. | | | | xTAG GPP positive results are presumptive and must be confirmed by FDA-cleared tests or other acceptable reference methods. | | | | The results of this test should not be used as the sole basis for diagnosis, treatment, or other patient management decisions. Confirmed 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 xTAG Gastrointestinal Pathogen Panel 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. xTAG GPP is not intended to monitor or guide treatment for C. difficile infections. | | | | The xTAG GPP is indicated for use with the Luminex MAGPIX instrument. | | Targets<br>Reported | Adenovirus 40/41, Campylobacter (C. jejuni, C. coli and C. lari only), Clostridium difficile (C. difficile) toxin A/B, Cryptosporidium (C. parvum and C. hominis only), Escherichia coli (E. coli) 0157, Enterotoxigenic Escherichia coli (ETEC) LT/ST, Entamoeba histolytica (E. histolytica), Giardia (G. lamblia only - also known as G. intestinalis and G. duodenalis), Norovirus GI/GII, Rotavirus A, Salmonella, Shiga-like Toxin producing E. coli (STEC) stx 1/stx 2, Shigella (S. boydii, S. sonnei, S. flexneri and S. dysenteriae), Vibrio cholerae (V. cholerae) | Campylobacter (C. jejuni, C. coli and C. lari only), Clostridium difficile (C. difficile) toxin A/B, Cryptosporidium (C. parvum and C. hominis only), Escherichia coli (E. coli) O157, Enterotoxigenic Escherichia coli (ETEC) LT/ST, Giardia (G. lamblia only - also known as G. intestinalis and G. duodenalis), Norovirus GI/GII, Rotavirus A, Salmonella, Shiga-like Toxin producing E. coli (STEC) stx 1/stx 2, Shigella (S. boydii, S. sonnei, S. flexneri and S. dysenteriae) | {8}------------------------------------------------ Image /page/8/Picture/0 description: The image shows the word "Luminex" in a bold, sans-serif font. The word is black, except for a red dot above the "i". The dot is slightly offset to the right. There is a registered trademark symbol to the right of the "x". ### xTAG® GPP with Luminex® MAGPIX® Traditional 510(k) Submission {9}------------------------------------------------ Image /page/9/Picture/1 description: The image shows the word "Luminex" in a bold, sans-serif font. The word is black, except for a red dot above the "i". There is a registered trademark symbol to the right of the "x". ## Standards/Guidance Documents referenced (if applicable): Table 3: Guidance Documents | | Title | Date | |----|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------| | 1 | Establishing the Performance Characteristics of In Vitro Diagnostic Devices for the Detection of Clostridium difficile | Nov. 29, 2010 | | 2 | Class II Special Controls Guidance Document: Norovirus Serological Reagents | Mar. 9, 2012 | | 3 | Class II Special Controls Guidance Document: Instrumentation for Clinical Multiplex Test Systems - Guidance for Industry and FDA Staff | Mar. 10, 2005 | | 4 | Guidance for the Content of Premarket Submissions for Software Contained in Medical Devices | May 11, 2005 | | 5 | Guidance document for Format for Traditional and Abbreviated 510(k)s | Aug. 12, 2005 | | 6 | Guidance on the CDRH Premarket Notification Review Program, 510(k) Memorandum #K86-3 | June 30, 1986 | | 7 | The New 510(k) Paradigm - Alternate Approaches to Demonstrating Substantial Equivalence in Premarket Notifications - Final Guidance | Mar. 20, 1998 | | 8 | The 510(k) Program: Evaluating Substantial Equivalence in Premarket Notifications [510(k)] | Dec. 27, 2011 | | 9 | Guidance for Industry and Food and Drug Administration Staff - eCopy Program for Medical Device Submissions | Oct. 10, 2013 | | 10 | Guidance for Industry and Food and Drug Administration Staff - FDA and Industry Actions on Premarket Notification (510(k)) Submissions: Effect on FDA Review Clock and Goals | Oct. 15, 2012 | #### Table 4: Standards | | Standard<br>No. | Recognition<br>Number<br>(FDA) | Standards Title | Date | |---|-----------------|--------------------------------|------------------------------------------------------------------------------------------------------------------------------------|------------| | 1 | EP05-A2 | 7-110 | Evaluation of Precision Performance of Quantitative<br>measurement Methods (2nd ed.) | 10/31/2005 | | 2 | EP07-A2 | 7-127 | Interference Testing in Clinical Chemistry (2nd<br>edition) | 05/21/2007 | | 3 | EP12-A2 | 7-152 | User Protocol for Evaluation f Qualitative Test<br>Performance (2nd edition) | 09/09/2008 | | 4 | EP14-A2 | 7-143 | Evaluation of Matrix Effects (2nd edition) | 03/16/2012 | | 5 | EP15-A2 | 7-153 | User Verification of Performance for Precision and<br>Trueness (2nd edition) | 09/09/2008 | | 6 | EP17-A | 7-194 | Protocol for Determination of Limits of Detection<br>and Limits of Quantitation<br>(NOTE: Original studies included this standard) | 03/28/2009 | | 7 | EP17-A2 | 7-233 | Evaluation of Detection Capability for Clinical<br>Laboratory Measurement Procedures | 01/15/2013 | | 8 | ISO 14971 | 5-40 | Application of Risk Management to Medical Devices | 08/20/2012 | | 9 | MM03-A2 | 7-132 | Molecular Diagnostic Methods for Infectious<br>Diseases (2nd edition) | 09/09/2008 | {10}------------------------------------------------ Image /page/10/Picture/0 description: The image shows the Luminex logo. The logo is black, except for a red dot above the "i" in Luminex. The logo is a sans-serif font and has a registered trademark symbol after the "x". xTAG® GPP with Luminex® MAGPIX® Traditional 510(k) Submission | Standard<br>No. | Recognition<br>Number<br>(FDA) | Standards Title | Date | | |-----------------|--------------------------------|-----------------|----------------------------------------------------------------|------------| | 10 | MM13-A | 7-191 | Collection, Transport, Preparation and Storage of<br>Specimens | 03/18/2009 | # Analytical Performance: The reagents tested in submission k121894 remain the same as the reagents used in testing performed towards this submission. Therefore, the study reports and results presented in the submission summary in k121894 for Analytical Reactivity, Carry-Over Contamination, Limit of Detection, Repeatability, Analytical Specificity (including Interference), Evaluation of Fresh vs. Frozen Stool, and Reproducibility / Precision are all still applicable to the new device. Results presented below for each of these studies are additive to results previously presented in k121894 and include results for Adenovirus 40/41, Entamoeba histolytica (E. histolytica), and Vibrio cholerae (V. cholerae) cholera toxin gene (ctx). This section of the summary includes updated results for these three analytes for the following studies: - 1. Analytical Reactivity - 2. Carry-over Contamination - 3. Limit of Detection - 4. Repeatability - 5. Analytical Specificity and Interference - 6. Evaluation of Fresh vs. Frozen Stool - 7. Reproducibility / Precision Additionally, a study demonstrating results of testing analytes in stool as compared to stool in Cary-Blair media is presented, at the limit of detection for each analyte to demonstrate that either sample type can be used with xTAG GPP. Finally, a summary of negative control failures and sample re-run rates for analytical performance studies is provided. {11}------------------------------------------------ Image /page/11/Picture/0 description: The image shows the word "Luminex" in a bold, sans-serif font. The word is black, except for a red dot above the "i". The registered trademark symbol is to the right of the "x". #### Analytical Reactivity Analytical reactivity was assessed through empirical testing of a wide range of clinically relevant GI pathogen strains, genotypes and isolates representing temporal and geographical diversity for each analyte. Through testing of unique samples covering the additional intended use pathogens, reactivity was established at concentrations 2 to 3 times the limit of detection. Adenovirus - The Limit of Detection (LoD) using Adenovirus 40, Zeptometrix 0810084CF (Dugan) and Adenovirus 41, Zeptometrix 0810085CF (Tak) were found to be 1.45E+01 TCID55/mL (or 4.89E+06 Copies/mL) and 7.69E+00 TCID5g/mL (or 1.48E+07 Copies/mL), respectively (see LoD section below). The following two samples were tested at the Centers for Disease Control and Prevention (CDC) (Atlanta, Georgia, USA). Note: these samples were different isolates of the strains used in the LoD study (See LoD section below). The amount of the viral target DNA for GP-093 and GP-094 was measured by real-time PCR and the Ct values generated were used to calculate the DNA copy number. The lowest reactivity titers for GP-093 and GP-094, were found to be at 3x and 1x multiple of LoD level, respectively. | Run Batch ID | Target | Source ID | Strain or Serotype | Reactivity<br>Titer<br>(Copies/mL) | Results Summary | |--------------------------|------------------|--------------|-----------------------------|------------------------------------|-----------------| | Analytical reactivity_II | Adenovirus<br>40 | CDC – GP-093 | Dugan<br>pCMK₂Gr₁₀, 9/23/91 | 1.49E+07 | POS | | Analytical reactivity_II | Adenovirus<br>41 | CDC – GP-094 | Tak<br>HeLa₂Gr₁₀, 9/23/91 | 1.43E+07 | POS | Table 5: Adenovirus Reactivity List Furthermore, in sequencing analysis of clinical specimens tested as part of the multi-site clinical study of xTAG GPP, 9 Adenovirus 40 and 28 Adenovirus 41 positive samples were detected by the assay and sequencing. | Target | Clinical Sample ID | |---------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------| | Adenovirus 40 | GPP03-092B, GPP03-099B, GPP03-101B, GPP03-102B, GPP03-103B, GPP03-106B, GPP03-109B,<br>GPP03-300B, GPP03-240B | | Adenovirus 41 | GPP03-001B, GPP03-003B, GPP03-007B, GPP03-013B, GPP03-014B, GPP03-019B, GPP03-020B,<br>GPP03-022B, GPP03-025B, GPP03-026B, GPP03-028B, GPP03-029B, GPP03-033B, GPP03-035B,<br>GPP03-036B, GPP03-037B, GPP03-038B, GPP03-039B, GPP03-048B, GPP03-055B, GPP03-060B,<br>GPP03-095B, GPP03-229B, GPP03-313B, GPP04-159, GPP04-174, GPP02-129, GPP02-192 | Table 6: Adenovirus Clinical Specimen Positive by the xTAG GPP Entamoeba histolytica - The LoD for Entamoeba histolytica, ATCC 30890 was found to be 2.88E+01 Cells/mL, equivalent to 4.30E+02 Copies/mL (see LoD section below). For E.histolytica, ATCC 50007, 50481, 50738 and 50454, the titer information expressed in Cells/mL could not be obtained. To standardize the quantification units for all E.histolytica strains, in this Analytical Reactivity study the amount of target DNA was measured by real-time PCR and the Ct values generated were used to calculate the DNA copy numbers. The reactivity titers for most of the strains were in the range of 0.4x to 6.7x multiple of LoD level for E.histolytica. The reactivity titer {12}------------------------------------------------ Image /page/12/Picture/0 description: The image shows the word "Luminex" in a bold, sans-serif font. The word is black, except for a red dot above the "i". There is a registered trademark symbol to the right of the "x". for ATCC 50738 (Rahman) was found to be 0.2x multiple of LoD level. Table 7: Entamoeba histolytica Reactivity List | Run Batch ID | Target | Source | Strain or Serotype | Reactivity Titer<br>(Cells or<br>Copies/mL) | Results<br>Summary | |---------------------------------|-----------------------------------------------------|-------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------|--------------------| | 20120216_JF_GPP_Reactivity_MP | Entamoeba<br>histolytica | ATCC 30015 | (HK-9, colonic biopsy<br>from adult human<br>male with amebic<br>dysentery, Korea);<br>frozen | 2.86E+00 Cells/mL,<br>or 1.82E+02<br>Copies/mL | POS | | 20120216_JF_GPP_Reactivity_MP | Entamoeba<br>histolytica | ATCC 30190 | (HB-301:NIH, feces<br>from adult human<br>male with amebic<br>dysentery, Burma,<br>1960); test tube | 1.07E+03<br>Copies/mL | POS | | 20120216_JF_GPP_Reactivity_MP | Entamoeba<br>histolytica | ATCC 30457 | (HU-21:AMC, colonic<br>biopsy from male<br>child with amebic<br>dysentery, Little<br>Rock, AR, 1970); test<br>tube | 1.68E+03<br>Copies/mL | POS | | 20120216_JF_GPP_Reactivity_MP | Entamoeba<br>histolytica | ATCC 30458 | (200:NIH); frozen | 1.83E+02 Cells/mL,<br>or 2.42E+03<br>Copies/mL | POS | | 20120216_JF_GPP_Reactivity_MP | Entamoeba<br>histolytica | ATCC 30459 | (HM-1:IMSS [ABRM];<br>feces from adult<br>human male,<br>asymptomatic cyst<br>passer, England,<br>1972); test tube | 1.83E+02 Cells/mL,<br>or 1.10E+03<br>Copies/mL | POS | | 20120314_JF_GPP_React_MP | Entamoeba<br>histolytica | ATCC 30889 | (H-458:CDC<br>[ATCC30217], feces<br>from human adult<br>female with amebic<br>dysentery, Asia (?),<br>(patient in U.S. for<br>treatment), 1971);<br>test tube | 8.78E+02<br>Copies/mL | POS | | 20120411_JF_GPP_React_MP | Entamoeba<br>histolytica | ATCC 30923 | (HU-2:MUSC) | 1.61E+03<br>Copies/mL | POS | | 20120207_JF_GPP_Reactivity_MP | Entamoeba<br>histolytica | ATCC 30925 | (HU-1:CDC, feces of<br>female child,<br>asymptomatic, sero-<br>negative cyst passer,<br>Cherokee, NC, 1978) | 1.89E+02<br>Copies/mL | POS | | 20120411_JF_GPP_React_MP | Entamoeba<br>histolytica | ATCC 50007 | DKB | 2.88E+03<br>Copies/mL | POS | | 20120411 JF GPP React MP | Entamoeba<br>histolytica | ATCC 50481 | SD157 | 1.36E+03<br>Copies/mL | POS | | 20120411 JF GPP React MP | Entamoeba<br>histolytica | ATCC 50738 | Rahman | 8.90E+01<br>Copies/mL | POS | | 20120411 JF GPP React MP | Entamoeba<br>histolytica | ATCC 50454 | HB-301:NIH | 1.08E+03<br>Copies/mL | POS | | Run Batch ID | Target | Source | Strain or Serotype | Reactivity Titer<br>(CFU/mL) | Results<br>Summary | | 20120827-JX-V cholera-AR-MP | Vibrio cholerae<br>Pacini | NCTC 30 | Non-O:1, ATCC<br>4735;MARTIN 1 | 6.00E+08 | NEG | | 20120827-JX-V cholera-AR-MP | Vibrio cholerae | NCTC 4714 | Non-O:1, Isolated<br>from pilgrims in El<br>Tor quarantine camp,<br>El Tor 34-D 19 | 6.00E+08 | NEG | | 20120827-JX-V cholera-AR-MP | Vibrio cholerae | NCTC 7260 | O:1, EGYPT 117 | 7.02E+06 | POS | | 20120827-JX-V cholera-AR-MP | Vibrio cholerae | NCTC 11500 | Non-O:1, VL 7050 | 6.00E+08 | NEG | | 20120827-JX-V cholera-AR-MP | Vibrio cholerae | NCTC 11507 | Non-O:1, VL 1941 | 6.00E+08 | NEG | | 20120827-JX-V cholera-AR-MP | Vibrio cholerae | NCTC 11510 | O:1, VL 01211 | 7.02E+06 | POS | | 20120827-JX-V cholera-AR-MP | Vibrio cholerae | NCTC 12945 | 0:139 (Non-O:1<br>(NAG) — reference<br>strain for 0:139<br>serovar | 7.02E+06 | POS | | 20120827-JX-V cholera-AR-MP | Vibrio cholerae | NCTC 12946 | 0:139 (Non-O:1<br>(NAG)) | 7.02E+06 | POS | | 20120406-JX-AnaReact-Vibrio2-MP | Vibrio cholerae<br>Pacini | ATCC 14033 | O:1, El Tor DO<br>1930;CN 5774;R.<br>Hugh 1092, Serotype<br>Inaba, Non-<br>toxinogenic | 1.50E+08 | NEG | | 20120404-JX-AnaReact-Vibrio-MP | Vibrio cholerae<br>asiaticae (Trevisan)<br>Pfeiffer | ATCC 14035 | O:1, Serotype Ogawa<br>[7787] | 7.02E+06 | POS | | 20120404-JX-AnaReact-Vibrio-MP | Vibrio cholerae<br>Pacini | ATCC 14101 | O:1, Serotype<br>Ogawa, clinical<br>specimen – human<br>([185754] cholera<br>epidemic circa 1960,<br>Calcutta) Calcutta<br>India | 7.02E+06 | POS | | 20120406-JX-AnaReact-Vibrio2-MP | Vibrio cholerae<br>Pacini | ATCC 14374 | Non-O:1 (NAG),<br>5035; R. Hugh 1513 | 1.50E+08 | NEG | | 20120921-MB-VibrioAnalytical-MP | Vibrio cholerae<br>Pacini | ATCC 14730 | Non-O:1 (Serovar<br>O:2), biovar El Tor,<br>Subgroup III of<br>Gardner and<br>Venkatraman, NCTC<br>4711, NANKING | 6.00E+08 | NEG | | Run Batch ID | Target | Source | Strain or Serotype | Reactivity Titer<br>(CFU/mL) | Results<br>Summary | | | | | 32/123 | | | | 20120921-MB-VibrioAnalytical-MP | Vibrio cholerae<br>Pacini | ATCC 14731 | Non-O:1, (Serovar<br>O:3), biovar El Tor,<br>Subgroup V of<br>Gardner and<br>Venkatraman, NCTC<br>4715, El Tor 34-D<br>23;CN 3426 | 7.02E+06 | NEG | | 20120921-MB-VibrioAnalytical-MP | Vibrio cholerae<br>Pacini | ATCC 14732 | Non-O:1 (Serovar<br>O:4), biovar El Tor,<br>Subgroup VI of<br>Gardner and<br>Venkatraman, NCTC<br>4716, KASAULI 73 | 6.00E+08 | NEG | | 20120921-MB-VibrioAnalytical-MP | Vibrio cholerae<br>Pacini | ATCC 14733 | Non-O:1 (Serovar<br>O:7), biovar El Tor,<br>Subgroup II of<br>Gardner and<br>Venkatraman, NCTC<br>8042, NANKING<br>32/124 | 6.00E+08 | NEG | | 20120404-JX-AnaReact-Vibrio-MP | Vibrio cholerae<br>Pacini | ATCC 25870 | O:1, Serotype Inaba | 7.02E+06 | POS | | 20120404-JX-AnaReact-Vibrio-MP | Vibrio cholerae<br>Pacini | ATCC 25872 | Non-O:1 (NAG),<br>Isolated from a<br>patient with clinical<br>cholera | 7.02E+06 | POS | | 20120404-JX-AnaReact-Vibrio-MP | Vibrio cholerae<br>Pacini | ATCC 25873 | Non-O:1 (NAG),<br>Isolated from a<br>patient with clinical<br>cholera | 7.02E+06 | POS | | 20120404-JX-AnaReact-Vibrio-MP | Vibrio cholerae<br>Pacini | ATCC 51394 | 0:139 (Non-O:1<br>[NAG]), Cholera<br>patient, Madras,<br>India | 7.02E+06 | POS | | 20120404-JX-AnaReact-Vibrio-MP | Vibrio cholerae<br>Pacini | ATCC 51395 | 0:139 (non 0:1<br>[NAG]), clinical<br>specimen - human<br>(cholera patient,<br>Madras, India) | 7.02E+06 | POS | | 20120404-JX-AnaReact-Vibrio-MP | Vibrio cholerae | ATCC BAA-<br>2163 | O:1, Isolated from a<br>patient in Artibonite<br>Department, Haiti,<br>October 2010,<br>Serotype Ogawa,<br>Biogroup El Tor<br>cholera toxin positive<br>CDC Isolate 2010 EL-<br>1786 | 7.02E+06 | POS | {13}------------------------------------------------ Image /page/13/Picture/0 description: The image shows the Luminex logo. The word "Luminex" is written in a bold, sans-serif font, with the "i" dotted with a red circle. The logo is simple and modern, and the red dot adds a pop of color. Vibrio cholerae - The LoD using Vibrio cholerae Pacini ATCC 14101 (serovar 0:1) was found to be 2.34E+06 CFU/mL. For this Analytical Reactivity study 3xLoD=7.02E+06 CFU/mL, and this was used for initial reactivity testing. In addition to toxinogenic strains, (i.e. 01 and 0139), the xTAG GPP assay also detects any non:01 Vibrio cholerae strains that do express cholera toxin gene, ctx (xTAG GPP Vibrio cholerae primers target gene), but not the non:01 strains that may cause clinical symptoms such as diarrhea by expressing a different virulence factor, which is likely the case for sample ATCC 14374 and other non:01 strains in this table. Both non-01 ATCC 25872 and non-O1 ATCC 25873 strains, were tested in sequencing assays and confirmed to contain the ctx gene with well conserved xTAG GPP Vibrio cholerae primer binding regions. #### Table 8: Vibrio cholerae Reactivity List {14}------------------------------------------------ Image /page/14/Picture/0 description: The image shows the logo for Luminex. The logo is black, with the word "Luminex" in a bold, sans-serif font. There is a red dot above the "i" in Luminex. The logo is simple and modern. #### xTAG® GPP with Luminex® MAGPIX® Traditional 510(k) Submission Table 9 summarizes the samples reactive with xTAG GPP. Note that in addition to toxinogenic strains, i.e. O1 and O139, xTAG GPP assay detects any non:O1 Vibrio cholerae strains that do express cholera toxin gene (xTAG GPP Vibrio cholerae primers target gene), but not the non:01 strains that may cause clinical symptoms such as diarrhea by expressing a different virulence {15}------------------------------------------------ # Luminex. factor, which is likely the case for ATCC 14374 and other non:01 strains tested in this study. Both non-O1 ATCC 25872 and non-O1 ATCC 25873 strains, were tested in sequencing assays and confirmed to contain the ctx gene with well conserved xTAG GPP Vibrio cholerae primer binding regions. Ten Vibrio cholerae strains that did not react with xTAG GPP assay are listed in Tables 8 and 10. | Pathogen | ATCC / Other<br>Reference | Pathogen | ATCC / Other<br>Reference | |---------------|---------------------------|----------------------------------------------------------------------------------------|---------------------------| | Adenovirus 40 | CDC - GP-093 | Adenovirus 41 | GPP03-095B | | Adenovirus 40 | GPP03-092B | Adenovirus 41 | GPP03-229B | | Adenovirus 40 | GPP03-099B | Adenovirus 41 | GPP03-313B | | Adenovirus 40 | GPP03-101B | Adenovirus 41 | GPP04-159 | | Adenovirus 40 | GPP03-102B | Adenovirus 41 | GPP04-174 | | Adenovirus 40 | GPP03-103B | Adenovirus 41 | GPP02-129 | | Adenovirus 40 | GPP03-106B | Adenovirus 41 | GPP02-192 | | Adenovirus 40 | GPP03-109B | Entamoeba histolytica | ATCC 30015 | | Adenovirus 40 | GPP03-240B | Entamoeba histolytica | ATCC 30190 | | Adenovirus 40 | GPP03-300B | Entamoeba histolytica | ATCC 30457 | | Adenovirus 41 | CDC - GP-094 | Entamoeba histolytica | ATCC 30458 | | Adenovirus 41 | GPP03-001B | Entamoeba histolytica | ATCC 30459 | | Adenovirus 41 | GPP03-003B | Entamoeba histolytica | ATCC 30889 | | Adenovirus 41 | GPP03-007B | Entamoeba histolytica | ATCC 30923 | | Adenovirus 41 | GPP03-013B | Entamoeba histolytica | ATCC 30925 | | Adenovirus 41 | GPP03-014B | Entamoeba histolytica | ATCC 50007 | | Adenovirus 41 | GPP03-019B | Entamoeba histolytica | ATCC 50481 | | Adenovirus 41 | GPP03-020B | Entamoeba histolytica | ATCC 50738 | | Adenovirus 41 | GPP03-022B | Entamoeba histolytica | ATCC 50454 | | Adenovirus 41 | GPP03-025B | Vibrio cholerae, serovar 0:1 | NCTC 7260 | | Adenovirus 41 | GPP03-026B | Vibrio cholerae, serovar 0:1 | NCTC 11510 | | Adenovirus 41 | GPP03-028B | Vibrio cholerae, serovar O:139 (Non-O:1<br>(NAG)) - reference strain for O:139 serovar | NCTC 12945 | | Adenovirus 41 | GPP03-029B | Vibrio cholerae, serovar O:139 (Non-O:1<br>(NAG)) | NCTC 12946 | | Adenovirus 41 | GPP03-033B | Vibrio cholerae asiaticae (Trevisan) Pfeiffer,<br>serovar 0:1, serotype Ogawa | ATCC 14035 | | Adenovirus 41 | GPP03-035B | Vibrio cholerae Pacini, serovar 0:1, Serotype<br>Ogawa | ATCC 14101 | | Adenovirus 41 | GPP03-036B | Vibrio cholerae Pacini, serovar 0:1, Serotype<br>Inaba | ATCC 25870 | | Adenovirus 41 | GPP03-037B | Vibrio cholerae Pacini, serovar Non-O:1<br>(NAG) | ATCC 25872 | | Adenovirus 41 | GPP03-038B | Vibrio cholerae Pacini, serovar Non-O:1<br>(NAG) | ATCC 25873 | | Adenovirus 41 | GPP03-039B | Vibrio cholerae Pacini, serovar O:139 (Non-<br>0:1 [NAG]) | ATCC 51394 | | Adenovirus 41 | GPP03-048B | Vibrio cholerae Pacini, serovar O:139 (Non-<br>0:1 [NAG]) | ATCC 51395 | | Adenovirus 41 | GPP03-055B | Vibrio cholera, serovar 0:1, serotype Ogawa,<br>biovar El Tor, cholera toxin positive | ATCC BAA-2163 | | Adenovirus 41 | GPP03-060B | | | Table 9: Reactivity of Adenovirus 40/41, Entamoeba histolytica and Vibrio cholerae Table 10: Vibrio cholerae Strains that did not React with xTAG GPP | Pathogen | ATCC / Other | Pathogen | ATCC / Other | |----------|--------------|----------|--------------| |----------|--------------|----------|--------------| {16}------------------------------------------------ Image /page/16/Picture/0 description: The image shows the logo for Luminex. The word "Luminex" is written in a bold, sans-serif font, with the letters in black. Above the "i" in Luminex is a red dot. To the right of the "x" is a registered trademark symbol. xTAG® GPP with Luminex® MAGPIX® Traditional 510(k) Submission | | Reference | | Reference | |-------------------------------------------------------------------------------------------|------------|---------------------------------------------------------------------------------------------------|------------| | Vibrio cholerae Pacini, Serovar<br>Non-O:1 (NAG) | NCTC 30 | Vibrio cholerae Pacini, Serovar Non-O:1 (NAG) | ATCC 14374 | | Vibrio cholerae, Serovar Non-O:1 | NCTC 4714 | Vibrio cholerae Pacini, Serovar O:2, biovar El<br>Tor, Subgroup III of Gardner and<br>Venkatraman | ATCC 14730 | | Vibrio cholerae, Serovar Non-O:1 | NCTC 11500 | Vibrio cholerae Pacini, Serovar O:3, biovar<br>ElTor, Subgroup V of Gardner and<br>Venkatraman | ATCC 14731 | | Vibrio cholerae, Serovar Non-O:1 | NCTC 11507 | Vibrio cholerae Pacini, Serovar O:4, biovar El<br>Tor; Subgroup VI of Gardner and<br>Venkatraman | ATCC 14732 | | Vibrio cholerae Pacini, Serovar 01,<br>biotype El Tor, serotype Inaba,<br>non-toxinogenic | ATCC 14033 | Vibrio cholerae Pacini, Serovar O:7, biovar El<br>Tor; Subgroup II of Gardner and Venkatraman | ATCC 14733 | ### Carry-over Contamination The likelihood of carry-over contamination events was initially assessed and presented in k121894 by testing 2 represen…