(2 days)
The Non-variola Orthopoxvirus Real-time PCR Primer and Probe Set is intended for the in vitro qualitative presumptive detection of non-variola Orthopoxvirus DNA extracted from human pustular rash specimens and viral cell culture lysates submitted to a Laboratory Response Network (LRN) reference laboratory. The assay detects non-variola Orthopoxvirus DNA, including Vaccinia, Cowpox, Monkeypox and Ectromelia viruses at varying concentrations. This assay does not differentiate Vaccinia virus or Monkeypox virus from other Orthopoxviruses detected by this assay and does not detect Variola virus. Refer to the CDC algorithm, Acute, Generalized Vesicular or Pustular Rash Illness Testing Protocol in the United States for recommended testing and evaluation algorithms for patients presenting with acute, generalized pustular or vesicular rash illness.
Results of this assay are for the identification of non-variola Orthopoxvirus DNA. These results must be used in conjunction with other diagnostic assays and clinical observations to diagnose Orthopoxvirus infection. The assay should only be used to test specimens with low/moderate risk of smallpox exists, viral culture should not be attempted. Negative results obtained with this device do not prection and should not be used as the sole basis for treatment or other patient management decisions.
Use is limited to Laboratory Response Network (LRN) designated laboratories.
The Non-variola Orthopoxvirus Real-time PCR Primer and Probe Set uses a fluorogenic probe, consisting of an oliqonucleotide 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 the Taq polymerase degrades the probe causing the reporter dye to separate from the quencher dye, thereby generating a fluorescent signal. With each cycle, additional reporter dye molecules are cleaved from their respective probes and the fluorescence intensity is monitored during the PCR in real-time. The Taq polymerase used in this assay is inactive at room temperature and is activated by incubation at 95°C, thus minimizing the production of nonspecific amplification products.
Each extracted DNA sample is tested using the Non-variola Orthopoxvirus Real-time PCR Primer and Probe set alonq with an internal control primer and probe set(s) to demonstrate adequate DNA extraction, proper function of common reagents and equipment, and the absence of inhibitory substances.
This document is a 510(k) summary for the Non-variola Orthopoxvirus Real-time PCR Primer and Probe Set, submitted by the Centers for Disease Control and Prevention (CDC). It is a re-submission (K221658) of a previously cleared device (K181205). The document largely states that many aspects of the device are unchanged from the previous submission and directs further inquiries about performance characteristics to the CDC.
Based on the provided text, many of the requested details are not explicitly stated in this FDA summary document. The document refers to an "Original Submission (K181205)" for comparison and states that for most performance characteristics, inquiries should be directed to the CDC. However, based on the information available:
1. A table of acceptance criteria and the reported device performance
The document does not explicitly state "acceptance criteria" in a tabulated format or provide detailed "reported device performance" in this summary. It mentions an "Analytical Limit of Detection (LoD)" and "Analytical Sensitivity and Specificity" studies were performed. It also indicates "Clinical Performance" studies were conducted, but the results for these are not presented here. Given that this is a re-submission with an "Unchanged" intended use and principle of operation, the acceptance criteria and performance would likely be similar to or supported by the original submission (K181205).
2. Sample size used for the test set and the data provenance (e.g. country of origin of the data, retrospective or prospective)
This information is not explicitly provided in the text. It states that inquiries regarding performance characteristics should be directed to the CDC.
3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts (e.g. radiologist with 10 years of experience)
This information is not explicitly provided in the text.
4. Adjudication method (e.g. 2+1, 3+1, none) for the test set
This information is not explicitly provided in the text.
5. If a multi reader multi case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance
This device is a PCR primer and probe set for detecting viral DNA, not an AI-assisted diagnostic device. Therefore, an MRMC comparative effectiveness study involving human readers and AI assistance is not applicable and was not performed.
6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done
This device is a molecular diagnostic assay. Its performance is inherent to the chemical reactions and detection system, and it operates "stand-alone" in the sense that the test result is directly outputted by the instrumentation without human real-time modification of the analytical process. However, human interpretation of the results and clinical context is required as stated in the Indications for Use: "Results of this assay are for the identification of non-variola Orthopoxvirus DNA. These results must be used in conjunction with other diagnostic assays and clinical observations to diagnose Orthopoxvirus infection."
7. The type of ground truth used (expert consensus, pathology, outcomes data, etc.)
The document refers to "identification of non-variola Orthopoxvirus DNA" as the output. The ground truth for such molecular tests typically involves:
- Validated reference strains/samples: For analytical sensitivity and specificity, the ground truth would be precisely known viral DNA concentrations or the known presence/absence of target and non-target organisms.
- Clinical diagnosis/outcome: For clinical performance, the ground truth would be established through a combination of other diagnostic assays, clinical observations, and potentially follow-up outcomes, as implied by the statement "These results must be used in conjunction with other diagnostic assays and clinical observations to diagnose Orthopoxvirus infection."
Specific details are not provided in this summary.
8. The sample size for the training set
This information is not provided in the text. As a PCR diagnostic kit, there isn't typically a "training set" in the machine learning sense. Instead, development involves assay design, optimization, and then validation studies.
9. How the ground truth for the training set was established
Not applicable as there is no "training set" in the machine learning context for this type of device. Assay optimization and validation (which could be considered analogous to "training" and "testing" in a broad sense for a traditional diagnostic) would rely on well-characterized samples with established viral content.
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Centers for Disease Control and Prevention Julie Villanueva Laboratory Preparedness and Response Branch Chief 1600 Clifton Road NE, MS: H24-11 Atlanta, Georgia 30329
June 10, 2022
Re: K221658
Trade/Device Name: Non-variola Orthopoxvirus Real-time PCR Primer and Probe Set Regulation Number: 21 CFR 866.3315 Regulation Name: Nucleic Acid Based Reagents For Detection Of Non-Variola Orthopoxviruses Regulatory Class: Class II Product Code: PBK Dated: June 7, 2022 Received: June 8, 2022
Dear Julie Villanueva:
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. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. 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 Part
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801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4. Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
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 https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.
For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely,
Noel J. Gerald, Ph.D. Branch Chief Bacterial Respiratory and Medical Countermeasures Branch Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health
Enclosure
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Indications for Use
510(k) Number (if known) K221658
Device Name
Non-variola Orthopoxvirus Real-time PCR Primer and Probe Set
Indications for Use (Describe)
The Non-variola Orthopoxvirus Real-time PCR Primer and Probe Set is intended for the in vitro qualitative presumptive detection of non-variola Orthopoxvirus DNA extracted from human pustular rash specimens and viral cell culture lysates submitted to a Laboratory Response Network (LRN) reference laboratory. The assay detects non-variola Orthopoxvirus DNA, including Vaccinia, Cowpox, Monkeypox and Ectromelia viruses at varying concentrations. This assay does not differentiate Vaccinia virus or Monkeypox virus from other Orthopoxviruses detected by this assay and does not detect Variola virus. Refer to the CDC algorithm, Acute, Generalized Vesicular or Pustular Rash Illness Testing Protocol in the United States for recommended testing and evaluation algorithms for patients presenting with acute, generalized pustular or vesicular rash illness.
Results of this assay are for the identification of non-variola Orthopoxvirus DNA. These results must be used in conjunction with other diagnostic assays and clinical observations to diagnose Orthopoxvirus infection. The assay should only be used to test specimens with low/moderate risk of smallpox exists, viral culture should not be attempted. Negative results obtained with this device do not prection and should not be used as the sole basis for treatment or other patient management decisions.
Use is limited to Laboratory Response Network (LRN) designated laboratories.
Type of Use (Select one or both, as applicable)
| Prescription Use (Part 21 CFR 801 Subpart D) |
|---|
| Over-The-Counter Use (21 CFR 801 Subpart C) |
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Centers for Disease Control and Prevention Non-variola Orthopoxvirus Real-time PCR Primer and Probe Set 510(k) Premarket Notification
510(k) Summary 10.
This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92.
| Assigned 510(k)number: | TBD |
|---|---|
| Submitted by: | Centers for Disease Control and Prevention1600 Clifton Road NEAtlanta, GA 30329 |
| Contact Person: | Julie Villanueva, PhDLaboratory Preparedness and Response Branch ChiefDivision of Preparedness and Emerging InfectionsNational Center for Emerging and Zoonotic InfectiousDiseasesCenters for Disease Control and Prevention(Registration number: 1050190)1600 Clifton Road, NE, MS H24-11Atlanta, GA 30329(404) 639-3851 (office)Jfv3@cdc.gov |
| Date prepared: | June 7, 2022 |
| Device trade name: | Non-variola Orthopoxvirus Real-time PCRPrimer and Probe Set |
| Classification name andregulation (if applicable): | 21 CFR 866.3315 |
| Predicate device(s): | Non-variola Orthopoxvirus Real-time PCRPrimer and Probe Set (K181205) |
Background
Variola virus, a member of the Orthopoxvirus genus, is the causative agent of smallpox and was certified eradicated in 1980 by the World Health Organization. At that time, smallpox vaccinations were ceased worldwide as a result. However, in recent years, concerns over the potential use of Variola virus as a biological weapon led the United States to resume smallpox vaccinations on a limited basis. Since the smallpox vaccine contains live Vaccinia virus, it is possible for vaccine recipients and/or their close contacts to develop adverse reactions to the vaccine including the emergence of pustules on the skin.
The Laboratory Response Network (LRN) is part of a national bioterrorism preparedness initiative created to ensure an effective laboratory response to biological threats by helping to improve the nation's public health laboratory infrastructure. Member laboratories must meet specific membership requirements and pass rigorous proficiency tests demonstrating their ability to accurately identify agents of concern. One of the major goals is the development and validation of rapid and specific assays for detection of
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biothreat agents and emerging infectious diseases. Accordingly, scientists at the Centers for Disease Control and Prevention have developed several realtime PCR based assays to detect non-variola Orthopoxvirus and other potential biothreat agents in an effort to meet the need for rapid detection.
The Non-variola Orthopoxvirus Real-time PCR Primer and Probe Set was developed for use in conjunction with clinical observations and other tests as described in the CDC algorithm, Acute, Generalized Vesicular or Pustular Rash Illness Testing Protocol in the United States. The assay is designed to aid in the identification of the causative aqent of a pustular or vesicular rash illness and to help rule out the presence of Variola virus in patients presenting with pustular rash illness.
This assay detects most commonly known human pathogenic Orthopoxyiruses (e.q., Vaccinia, Cowpox, and Monkeypox viruses) but does not detect Variola virus, the causative agent of smallpox. Vaccinia virus infection in the United States usually occurs in conjunction with smallpox vaccination or contact with a smallpox vaccine recipient. Monkeypox and Cowpox viruses are endemic to locations outside the United States, with the exception of the 2003 monkeypox outbreak associated with prairie dogs, which became infected due to imported African rodents.
On May 17, 2022, skin lesions that had several characteristics similar to monkevpox on a U.S. resident prompted specialized LRN testing of swab specimens collected from the resident; preliminary testing confirmed the presence of DNA consistent with an orthopoxvirus using the Non-variola Orthopoxvirus Real-time PCR Primer and Probe Set. This group of viruses includes monkeypox virus (the causative agent of monkeypox). Testing at CDC on May 18 confirmed the patient was infected with a strain from the West African clade of monkeypox virus. Additional cases of orthopoxvirus infection were subsequently detected using the Non-variola Orthopoxvirus Real-time PCR Primer and Probe Set.
Device Description
With the exception of some reagents and instrumentation, the Non-variola Orthopoxvirus Real-time PCR Primer and Probe Set device description remains unchanged from the 2018 submission (K181205).
The Non-variola Orthopoxvirus Real-time PCR Primer and Probe Set uses a fluorogenic probe, consisting of an oliqonucleotide 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 the Taq polymerase degrades the probe causing the reporter dye to separate from the quencher dye, thereby generating a fluorescent signal. With each cycle, additional reporter dye molecules are cleaved from their respective probes and the fluorescence intensity is monitored during the PCR in real-time. The Taq polymerase used in this assay is inactive at room temperature and is activated by incubation at 95°C, thus minimizing the production of nonspecific amplification products.
Each extracted DNA sample is tested using the Non-variola Orthopoxvirus Real-time PCR Primer and Probe set alonq with an internal control primer and
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probe set(s) to demonstrate adequate DNA extraction, proper function of common reagents and equipment, and the absence of inhibitory substances.
Intended Use
The Non-variola Orthopoxvirus Real-time PCR Primer and Probe Set is intended for the in vitro qualitative presumptive detection of non-variola Orthopoxvirus DNA extracted from human pustular or vesicular rash specimens and viral cell culture lysates submitted to a Laboratory Response Network (LRN) reference laboratory. The assay detects non-variola Orthopoxvirus DNA, including Vaccinia, Cowpox, Monkeypox and Ectromelia viruses at varying concentrations. This assay does not differentiate Vaccinia virus or Monkeypox virus from other Orthopoxviruses detected by this assay and does not detect Variola virus. Refer to the CDC algorithm, Acute, Generalized Vesicular or Pustular Rash Illness Testing Protocol in the United States for recommended testing and evaluation algorithms for patients presenting with acute, generalized pustular or vesicular rash illness.
Results of this assay are for the presumptive identification of non-variola Orthopoxvirus DNA. These results must be used in conjunction with other diagnostic assays and clinical observations to diagnose Orthopoxvirus infection. The assay should only be used to test specimens with low/moderate risk of smallpox. If a high risk of smallpox exists, viral culture should not be attempted. Negative results obtained with this device do not preclude Variola virus infection and should not be used as the sole basis for treatment or other patient management decisions.
Use is limited to Laboratory Response Network (LRN) designated laboratories.
Device Comparison
The following table summarizes the similarities and differences between the cleared assay and the new submission for this device.
| New Submission | Original Submission | |
|---|---|---|
| Device | Non-variola OrthopoxvirusReal-time PCR Primer andProbe Set | Non-variola OrthopoxvirusReal-time PCR Primer andProbe Set (K181205) |
| Intended Use | Unchanged | The Non-variolaOrthopoxvirus Real-timePCR Primer and Probe Setis intended for the in vitroqualitative presumptivedetection of non-variolaOrthopoxvirus DNAextracted from humanpustular or vesicular rashspecimens and viral cellculture lysates submittedto a Laboratory ResponseNetwork (LRN) reference |
| laboratory. The assay | ||
| detects non-variola | ||
| Orthopoxvirus DNA, | ||
| including vaccinia, | ||
| cowpox, monkeypox and | ||
| ectromelia viruses at | ||
| varying concentrations. | ||
| This assay does not | ||
| differentiate vaccinia virus | ||
| or monkeypox virus from | ||
| other orthopoxviruses | ||
| detected by this assay | ||
| and does not detect | ||
| variola virus. Refer to the | ||
| CDC algorithm, Acute,Generalized Vesicular or | ||
| Pustular Rash Illness | ||
| Testing Protocol in the | ||
| United States for | ||
| recommended testing and | ||
| evaluation algorithms for | ||
| patients presenting with | ||
| acute, generalized | ||
| pustular or vesicular rash | ||
| illness. | ||
| Results of this assay are | ||
| for the presumptiveidentification of non- | ||
| variola Orthopoxvirus | ||
| DNA. These results must | ||
| be used in conjunction | ||
| with other diagnostic | ||
| assays and clinical | ||
| observations to diagnose | ||
| Orthopoxvirus | ||
| infection. The assay | ||
| should only be used to | ||
| test specimens withlow/moderate risk of | ||
| smallpox. If a high risk of | ||
| smallpox exists, viral | ||
| culture should not be | ||
| attempted. Negative | ||
| results obtained with this | ||
| device do not preclude | ||
| Variola virus infection and | ||
| should not be used as the | ||
| sole basis for treatment | ||
| or other patient | ||
| management decisions. | ||
| Use is limited to | ||
| Laboratory ResponseNetwork (LRN) | ||
| designatedlaboratories. | ||
| Principle ofOperation | Unchanged | Nucleic acid amplificationand fluorescent probedetection |
| Sample Types | Unchanged | • Vesicle fluid, skin,crust, "roof"• Dry or wet swab oflesion (dry swab ispreferred)• Touch prep (slide) oflesion• Fresh biopsy of pustuleor vesicle (no formalin)• Viral cell culture lysates |
| Instrumentationand Software | Unchanged | Real-time PCRinstrumentation andsoftware |
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Establishment of Performance Characteristics
Inquiries regarding performance characteristics for the Non-variola Orthopoxvirus Real-time PCR Primer and Probe Set should be directed to the Centers for Disease Control and Prevention.
Analytical Limit of Detection (LoD)
The limit of detection for the Non-variola Orthopoxvirus Real-time PCR Primer and Probe Set was determined through an analytical sensitivity study.
Analytical Sensitivity and Specificity
Inquiries regarding performance characteristics for the Non-variola Orthopoxvirus Real-time PCR Primer and Probe Set should be directed to the Centers for Disease Control and Prevention.
Clinical Performance
Inquiries regarding clinical performance characteristics for the Non-variola Orthopoxvirus Real-time PCR Primer and Probe Set should be directed to the Centers for Disease Control and Prevention.
N/A