ANTHRACIS REAL-TIME PCR ASSAY

{0}------------------------------------------------ #### 510(k) Summary This summary of 510(k) substantial equivalence information is being submitted in accordance with the requirements of 21 CFR 807.92. | Assigned 510(k)<br>number: | K140426 | |----------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------| | Submitted by: | Centers for Disease Control and Prevention<br>1600 Clifton Road NE<br>Atlanta, GA 30333 | | Contact Person: | CDR Yon Yu, Pharm.D.<br>Associate Director for Regulatory Affairs<br>Office of the Director<br>National Center for Emerging and Zoonotic Infectious Diseases<br>Centers for Disease Control and Prevention<br>(Registration number: 1050190)<br>1600 Clifton Road, NE, MS C-18<br>Atlanta, GA 30333<br>(404) 639-3046 (office)<br>(404) 639-1275 (fax)<br>Fkb8@cdc.gov | | Date prepared: | February 14, 2014 | | Device trade name: | <i>B. anthracis</i> Real-time PCR Assay | | Classification name<br>and regulation: | <i>In vitro</i> diagnostic device for <i>Bacillus spp.</i><br>detection; 21 CFR 866.3045 | | Product Code: | NHT | | Class: | II | | Panel: | Microbiology (83) | | Predicate<br>device(s): | JBAIDS Anthrax Detection System<br>(K051713, K071188) November 18, 2005 | #### Background Anthrax is a zoonotic disease caused by B. anthracis that is transmissible to humans through handling or consumption of contaminated animal products. Infection can also occur through inhalation of B. anthracis spores from contaminated animal products such as wool or hides. Infection caused by human-to-human contact has been reported only rarely, and only via the cutaneous route (Versalovic, 2011). There have been 3 major presentations of anthrax in humans: cutaneous, ingestion, and inhalation. In cases of {1}------------------------------------------------ cutaneous anthrax, patients typically present with a painless blister or skin ulcer with a black area in the center. Inhalation anthrax is typically associated with cold or flu-like symptoms, cough, chest discomfort, shortness of breath, fatigue, and muscle aches. Symptoms of gastrointestinal anthrax typically include nausea, loss of appetite, bloody diarrhea, fever and severe stomach pain. Prior to the development of the LRN B. anthracis Real-time PCR Assay, identification of B. anthracis was determined by using phenotypic differences between B. anthracis and the rest of the B. cereus group. (i.e. lack of motility and hemolysis, susceptibility to penicillin, colony morphology, susceptibility to lysis by gamma phage) (Hoffmaster, 2002). However, these methods require growth of the microorganism and can take at least 24 hours incubation to obtain a result. Due to the prevalence of B. anthracis in the environment, and its past use as a biological weapon, it has long been an organism of concern. The use of B. anthracis in the bioterrorism attacks of 2001 resulting in cases of inhalation and cutaneous anthrax increased public health concern and reinforced the worry that it would be used in the same way again. For these reasons, there was a need for rapid testing to aid in the identification of B. anthracis. The Laboratory Response Network (LRN) is part of a national bioterrorism preparedness initiative and one of the major goals of this initiative is the development and validation of rapid and specific assays for agents likely to be used in a bioterrorism event. Accordingly, scientists at the Centers for Disease Control and Prevention have developed several real-time PCR based assays to detect B. anthracis and other potential agents of bioterrorism in an effort to meet the need for rapid detection. #### Device description The B. anthracis Real-time PCR Assay uses a fluorogenic probe, consisting of an oligonucleotide with a reporter dye (FAM) attached to the 5' end and a quencher dye (BHQ1) attached at or near the 3' end. The probe anneals to a specific target sequence located between the forward and reverse primers. During the extension phase of the PCR cycle, the 5′ nuclease activity of Tag polymerase degrades the probe causing the reporter dye to separate from the quencher dye and a fluorescent signal is generated. With each cycle, additional reporter dye molecules are cleaved from their respective probes and the fluorescence intensity is monitored during the PCR. The Tag polymerase used in this assay is inactive at room temperature. It must be activated by incubation at 95°C, which also minimizes the production of nonspecific amplification products. Each extracted DNA sample is tested with three B. anthracis primer and probe sets run as individual reactions. The primer and probe sets target genes encoding virulence factors as well as conserved regions of DNA from the B. anthracis chromosome. All three primer and probe sets must be positive for the overall result of the B. anthracis Real-time PCR Assay to be interpreted as positive. Any result that is positive for some, but not all three target reqions, {2}------------------------------------------------ is still considered equivocal and follow-up laboratory investigation should be performed per the LRN Bacillus anthracis Testing Algorithm. #### Intended Use The B. anthracis Real-time PCR Assay is an in vitro diagnostic test for the qualitative detection of plasmid and chromosomal DNA sequences from B. anthracis. The assay can be used to test human respiratory samples, whole blood, serum, plasma, swabs from lesions, CSF, pleural fluid, and bacterial culture isolates from individuals suspected of having anthrax. Results generated from direct specimen testing are presumptive for the identification of B. anthracis. Results generated from culture isolate testing should be used in conjunction with other conventional methods for identification of Bacillus anthracis isolates as part of the LRN Bacillus anthracis Testing Algorithm. The diagnosis of anthrax infection must be made based on history, signs, symptoms, exposure likelihood, and other laboratory evidences in addition to the identification of B. anthracis from cultures or detection directly in clinical specimens. #### Use is limited to Laboratory Response Network (LRN) designated laboratories. The B. anthracis Real-time PCR Assay is also intended for environmental specimen testing for biothreat detection and response. FDA has not evaluated claims related to the use of this assay on environmental specimens. #### Device comparison As previously mentioned, there are several culture based methods used for identification of B. anthracis. While they are reliable methods, they do not offer a fast result since a pure culture of the microorqanism must first be isolated, then set up with each test (gamma phage, morphology, motility, penicillin resistance, etc.). Each of these tests requires approximately 24 hours incubation before they can be interpreted. There is a currently marketed device that uses similar nucleic acid amplification and fluorescent probe detection technology called the JBAIDS Anthrax Detection System (Idaho Technology, Inc., 510(k) #K051713). The following table summarizes the similarities and differences between the two devices. {3}------------------------------------------------ | Device<br>(Owner) | <i>B. anthracis</i> Real-time PCR Assay<br>(Centers for Disease Control and<br>Prevention) | JBAIDS Anthrax Detection<br>System (Idaho Technology, Inc.) | |-------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------| | Similarities | | | | Intended<br>Use | The <i>B. anthracis</i> Real-time PCR Assay<br>is an <i>in vitro</i> diagnostic test for the<br>qualitative detection of plasmid and<br>chromosomal DNA sequences from <i>B.</i><br><i>anthracis</i> . The assay can be used to<br>test human respiratory samples, whole<br>blood, serum, plasma, swabs from<br>lesions, CSF, pleural fluid, and<br>bacterial culture isolates from<br>individuals suspected of having<br>anthrax.<br><br>Results generated from direct<br>specimen testing are presumptive for<br>the identification of <i>B. anthracis</i> .<br><br>Results generated from culture isolate<br>testing should be used in conjunction<br>with other conventional methods for<br>identification of <i>Bacillus anthracis</i><br>isolates as part of the LRN <i>Bacillus</i><br><i>anthracis</i> Testing Algorithm. The<br>diagnosis of anthrax infection must be<br>made based on history, signs,<br>symptoms, exposure likelihood, and<br>other laboratory evidences in addition<br>to the identification of <i>B. anthracis</i><br>from cultures or detection directly in<br>clinical specimens. | The JBAIDS Anthrax Detection<br>System is a real-time<br>polymerase chain reaction (PCR)<br>test system intended for the<br>qualitative <i>in vitro</i> diagnostic<br>(IVD) detection of target DNA<br>sequences on the pXO1 plasmid<br>(Target 1) and the pXO2 plasmid<br>(Target 2) from <i>Bacillus</i><br><i>anthracis</i> . The system can be<br>used to test human whole blood<br>collected in sodium citrate from<br>individuals suspected of having<br>anthrax, positive blood cultures,<br>and cultured organisms grown on<br>blood agar plates. The JBAIDS<br>Anthrax Target 2 assay is used<br>as a supplementary test only<br>after a positive result with the<br>Target 1 Assay.<br><br>The JBAIDS Anthrax Target 1<br>and Target 2 Assays are run on<br>the JBAIDS instrument using the<br>Diagnostic Wizard.<br><br>Results are for the presumptive<br>identification of <i>B. anthracis</i> , in<br>conjunction with culture and<br>other laboratory tests. The<br>following considerations also<br>apply: | | | Use is limited to Laboratory<br>Response Network (LRN)<br>designated laboratories.<br><br>The <i>B. anthracis</i> Real-time PCR<br>Assay is also intended for<br>environmental specimen testing for<br>biothreat detection and response.<br>FDA has not evaluated claims related<br>to the use of this assay on<br>environmental specimens. | The diagnosis of anthrax<br>infection must be made<br>based on history, signs,<br>symptoms, exposure<br>likelihood, other<br>laboratory evidence, in<br>addition to the<br>identification of pXO1 and<br>pXO2 targets either from<br>cultures or from direct<br>blood specimens. The assays have not been<br>evaluated with blood from<br>individuals without clinical<br>signs or symptoms who | ﺍﯾﻦ ﻣﯿﻼﺩﯼ ﺍﺯ ﻣﺮﮐﺰ ﺩﺭ ﺍﺳﺖ ﻣﯽ ﺍﺳﺖ. ﺍﯾﻦ ﻣﺮﮐﺰ ﺍﯾﻦ ﻣﯽ ﺍﺳﺖ. . . : · . ... : . ' . : : ・・ . : {4}------------------------------------------------ | Principle of Operation | Nucleic acid amplification and fluorescent probe detection | |------------------------|------------------------------------------------------------| |------------------------|------------------------------------------------------------| - were presumed exposed and who subsequently developed anthrax (inhalation or other forms of the disease), or from individuals with any form of anthrax (inhalational, cutaneous, or gastrointestinal). - The level of plasmid targets that would be present in blood from individuals with early systemic infection is unknown. - The definitive identification of B. anthracis from colony growth, liquid blood culture growth, or from blood specimens requires additional testing and confirmation procedures in consultation with public health or other authorities for whom reports are required. - The safety and effectiveness of other types of tests or sample types (not identified as "For in vitro diagnostic use") have not been established. | Nucleic acid amplification and fluorescent probe detection | |------------------------------------------------------------| |------------------------------------------------------------| . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 제 사용 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 14 - 1 ﺮ ﻣﺤﻤﺪ ﺍﻟﻤﻮﺿﻮﻉ ﺍﻟﻤﺴﺎﺣﺔ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪ , ર-ર {5}------------------------------------------------ | Differences | | | |-----------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------| | Sample Types | Swabs from lesions and vesicular material Whole blood (EDTA or sodium citrate) Serum/Plasma Respiratory specimens (transtracheal aspirates, bronchial lavage, and sputum) Cerebrospinal fluid Pleural fluid Bacterial culture isolates Environmental samples collected for investigational or surveillance use | Whole blood (sodium citrate) | | Instrumentation | Applied Biosystems 7500 Fast Dx Real-Time PCR System and Cepheid SmartCycler I or II Instruments with native software | JBAIDS integrated thermocycler and fluorimeter with Diagnostic Wizard software | | Targets | <i>B. anthracis</i> virulence plasmids and chromosomal region DNA | <i>B. anthracis</i> virulence plasmids | #### Establishment of Performance Characteristics - - - - - Inquiries regarding performance characteristics for the B. anthracis Real-time PCR Assay should be directed to the Centers for Disease Control and Prevention. #### Limit of Detection (LoD) The limit of detection for the B. anthracis Real-time PCR Assay was determined through in-house and multicenter sensitivity studies. #### Analytical Sensitivity and Specificity Inquiries regarding performance characteristics for the B. anthracis Real-time PCR Assay should be directed to the Centers for Disease Control and Prevention. {6}------------------------------------------------ # Clinical Performance Clinical performance of the B. anthracis Real-time PCR Assay was established through three evaluations: a) Evaluation of the BA3 marker to identify B. anthracis in a historical collection of Bacillus sp. isolates; b) Testing of known B. cereus isolates using the B. anthracis Real-time PCR Assay; c) Testing of known bacterial isolates using the B. anthracis Real-time PCR Assay. All three data sets compared the performance of the B. anthracis Real-time PCR Assay to a battery of tests performed by the CDC, including the gamma phage lysis, conventional culture, microbiological, and biochemical testing. Repeatability in clinical matrices was determined through a multicenter study. {7}------------------------------------------------ Image /page/7/Picture/0 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized depiction of an eagle or bird-like figure with outstretched wings. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular fashion around the bird-like figure. #### DEPARTMENT OF HEALTH & HUMAN SERVICES Public Health Service May 22, 2014 Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G60 Silver Spring, MD 20993-0002 CENTERS FOR DISEASE CONTROL AND PREVENTION YON YU ASSOCIATE DIRECTOR FOR REGULATORY AFFAIRS LABORATORY PREPAREDNESS AND RESPONSE BRANCH NCEZID/DPEI 1600 CLIFTON RD. NE, MS C-18 ATLANTA, GA 30333 USA Re: K140426 Trade/Device Name: B. anthracis Real-Time PCR Assay Regulation Number: 21 CFR 866.3045 Regulation Name: In vitro Diagnostic Devices for Bacillus spp. Detection Regulatory Class: II Product Code: NHT Dated: February 14, 2014 Received: February 25, 2014 Dear Ms. Yu: 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 {8}------------------------------------------------ Page 2-Ms. Yu 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 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/MedicalDevices/ResourcesforYou/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR 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/MedicalDevices/ResourcesforYou/Industry/default.htm. Sincerely yours, Sally A. Hojvat, M. Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological I-lealth Center for Devices and Radiological Health Enclosure {9}------------------------------------------------ #### Indications for Use #### K140426 ### Device Name B. anthracis Real-Time PCR Assay Indications for Use (Describe) The B. anthracis Real-Time PCR Assay is an in vitro diagnostic test for the qualitative detection of plasmid and chromosomal DNA sequences from B. anthracis. The assay can be used to test human respiratory samples, whole blood, serum, plasma, swabs from lesions, CSF, pleural fluid, and bacterial culture isolates from individuals suspected of having anthrax. Results generated from direct specimentive for the identification of B. anthracis. Results generated from culture isolate testing should be used in conventional methods for identification of Bacillus anthracis isolates as part of the LRN Bacillus anthracis Testing Algorithm. The diagnosis of anthrax infection must be made based on history, signs re likelihood, and other laboratory evidences, in addition to the identification of B. anthracis from cultures or detectly in clinical specimens. #### Use is limited to Laboratory Response Network (LRN) designated laboratories. The B. anthracis Real-time PCR Assay is also intended for environmental speciment testing for biothreat detection and response. FDA has not evaluated claims related to the use of this assay on environmental specimens. ## Type of Use (Select one or both, as applicable) 2 Prescription Use (Part 21 CFR 801 Subpart D) Over-The-Counter Use (21 CFR 801 Subpart C) Form Approved: OMB No. 0910-0120 Expiration Date: January 31, 2017 See PRA Statement on last page. #### 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) # John Hobson -S 2014.05.22 09:46:42 -04'00' {10}------------------------------------------------ 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 comblete 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." .