(299 days)
P160023
P160023/R004
No
The document describes a nucleic acid amplification test and automated system for detecting and quantifying HCV RNA. The changes described relate to reagent handling and a process control for detecting a faulty LED, which are not indicative of AI/ML. The data analysis mentioned is for verifying the new process control, not for training or validating an AI/ML model.
No
The device is an in vitro diagnostic (IVD) assay designed to detect and quantify hepatitis C virus (HCV) RNA. It aids in the diagnosis and management of HCV infection by providing information about viral load, but it does not directly treat or prevent a disease, which is the definition of a therapeutic device.
Yes
Explanation: The "Intended Use / Indications for Use" section explicitly states that "The Aptima HCV Quant Dx assay is indicated for use as an aid in the diagnosis of active HCV infection" and "as an aid in the management of HCV infected patients undergoing HCV antiviral drug therapy." These uses directly align with the definition of a diagnostic device.
No
The device description explicitly states that the changes are updates to the Assay Definition Module (ADM) and allow for additional reagent cycling on the Panther System. It also mentions that the Panther system is an integrated hardware and software system. This indicates that the device is not solely software, but rather a system that includes both hardware and software components.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The document explicitly states the intended use is for "detection and quantitation of hepatitis C virus (HCV) RNA in fresh and frozen human serum and plasma". This involves testing human specimens in vitro (outside the body) to provide information for diagnosis and management of a disease.
- Indications for Use: The indications clearly describe how the assay is used as an "aid in the diagnosis of active HCV infection" and as an "aid in the management of HCV infected patients undergoing HCV antiviral drug therapy". These are typical applications for IVD devices.
- Device Description: The description details a laboratory test that analyzes biological samples (serum and plasma) using a specific technology (real-time transcription-mediated amplification) to detect and measure a biological marker (HCV RNA). This is the core function of an IVD.
- Specimen Types: The document specifies the use of "human serum and plasma", which are common biological specimens used in IVD testing.
- Laboratory Setting: The description mentions the use of the "Panther system for automated specimen processing, and quantitation", indicating a laboratory or clinical setting where IVD tests are performed.
The entire context of the document, from the intended use to the technical description and performance studies, aligns with the definition and characteristics of an In Vitro Diagnostic device.
N/A
Intended Use / Indications for Use
The Aptima HCV Quant Dx assay is a real-time transcription-mediated amplification (TMA) test used for both detection and quantitation of hepatitis C virus (HCV) RNA in fresh and frozen human serum and plasma from HCV-infected individuals.
Plasma may be prepared in ethylenediaminetetraacetic acid (EDTA), anticoagulant citrate dextrose (ACD) solution, and plasma preparation tubes (PPT). Serum may be prepared in serum tubes and serum separator tubes (SST). Specimens are tested using the Panther system for automated specimen processing, amplification, detection, and quantitation. Specimens containing HCV genotypes 1 to 6 are validated for detection and quantitation in the assay.
The Aptima HCV Quant Dx assay is indicated for use as an aid in the diagnosis of active HCV infection in the following populations: individuals with antibody evidence of HCV infection with evidence of liver disease, individuals suspected to be actively infected with HCV antibody evidence, and individuals at risk for HCV infection with antibodies to HCV. Detection of HCV RNA indicates that the virus is replicating and, therefore, is evidence of active infection. Detection of HCV RNA does not discriminate between acute and chronic states of infection.
The Aptima HCV Quant Dx assay is also indicated for use as an aid in the management of HCV infected patients undergoing HCV antiviral drug therapy. The assay can be used to measure HCV RNA levels periodically prior to, during, and after treatment to determine sustained virological response (SVR) or nonsustained virological response (NSVR). Assay performance characteristics have been established for individuals infected with HCV and treated with certain direct-acting antiviral agents (DAA) regimens. No information is available on the assay's predictive value when other therapies are used. The results from the Aptima HCV Quant Dx assay must be interpreted within the context of all relevant clinical and laboratory findings.
The Aptima HCV Quant Dx assay is not approved for use as a screening test for the presence of HCV RNA in blood or blood products.
Product codes
MZP
Device Description
Clearance of this pre-market application will 1) enable additional reagent on-board / off-board cycling, allowing operators to load reagents onto the Panther System 8 times instead of 5 times; and 2) update the Assay Definition Module (ADM) to detect, flag and invalidate results impacted by a faulty or flickering LED, using estimated background minimum limits. These changes do not introduce any changes to the original design, method of manufacture, assay procedure, principle of operation, mechanism of action, conditions of use or hardware of the Panther instrument, or to the results interpretation for the cleared assay.
The Aptima HCV Quant Dx assay is a nucleic acid amplification test that uses real-time TMA technology to detect and quantitate HCV RNA for aiding diagnosis or to establish baseline viral load, as well as to measure on-treatment and post-treatment responses. The assay targets a conserved region of the HCV genome, detecting and quantitating genotypes 1, 2, 3, 4, 5, and 6. The assay is standardized against the 2nd WHO International Standard for Hepatitis C Virus (NIBSC Code 96/798).
The Aptima HCV Quant Dx assay involves three main steps, which all take place in a single tube on the Panther system: target capture, target amplification by TMA, and detection of the amplification products (amplicon) by the fluorescent labeled probes (torches).
During target capture, viral RNA is isolated from specimens. The specimen is treated with a detergent to solubilize the viral envelope, denature proteins, and release viral genomic RNA. Capture oligonucleotides hybridize to highly conserved regions of HCV RNA, if present, in the test specimen. The hybridized target is then captured onto magnetic microparticles that are separated from the specimen in a magnetic field. Wash steps remove extraneous components from the reaction tube.
Target amplification occurs via TMA, which is a transcription-mediated nucleic acid amplification method that utilizes two enzymes, Moloney murine leukemia virus (MMLV) reverse transcriptase and T7 RNA polymerase. The reverse transcriptase is used to generate a DNA copy (containing a promoter sequence for T7 RNA polymerase) of the target sequence. T7 RNA polymerase produces multiple copies of RNA amplicon from the DNA copy template. The Aptima HCV Quant Dx assay utilizes the TMA method to amplify a portion of the 5' UTR of the HCV genome. Amplification of this region is achieved using specific primers which are designed to amplify HCV genotypes 1, 2, 3, 4, 5, and 6.
Detection is achieved using single-stranded nucleic acid torches that are present during the amplification of the target and that hybridize specifically to the amplicon in real time. Each torch has a fluorophore and a quencher. When the torch is not hybridized to the amplicon, the quencher is in close proximity of the fluorophore and suppresses the fluorescence. When the torch binds to the amplicon, the quencher is moved farther away from the fluorophore and it will emit a signal at a specific wavelength when excited by a light source. As more torches hybridize to amplicon a higher fluorescent signal is generated. The time taken for the fluorescent signal to reach a specified threshold is proportional to the starting HCV concentration. Each reaction has an internal calibrator/internal control (IC) that controls for variations in specimen processing, amplification, and detection. The concentration of a sample is determined by the Panther system software using the HCV and IC signals for each reaction and comparing them to calibration information.
The reagents required to perform the Aptima HCV Quant Dx assay are available in 100-test kits. The kits are packaged in 3 boxes containing 11 reagents which are required for sample processing.
The Aptima HCV Quant Dx assay has been designed for and validated on the Panther system. The Panther system is an integrated hardware and software system that together with the Aptima HCV Quant Dx assay fully automates all the steps necessary to perform the assay from sample preparation through amplification of nucleic acid, detection, data reduction and amplicon inactivation.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
Not Found
Indicated Patient Age Range
Not Found
Intended User / Care Setting
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Description of the training set, sample size, data source, and annotation protocol
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Description of the test set, sample size, data source, and annotation protocol
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Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Intended Use Testing
Results from the Aptima HCV Quant Assay on Panther Fusion System System Software 7.2.9 were compared to those from the previous software version, 7.2.7 (reported to the FDA in the 2022 PMA annual report for the Aptima HCV Quant Dx Assay, P160023/R004).
The results met established acceptance criteria, confirming that assay performance is equivalent to the prior comparators, and demonstrating that the system is capable of performing the Aptima HCV Quant Dx Assay version 5.3.5.1 on the Panther Fusion System Software 7.2.9.
Sample Type: QC Control A; Target Conc (log IU/mL): Negative; Valid N: 120; Expected Result: Not Detected; Allowable Number of Outliers: 1 False positive; Standard Deviation: N/A; Result: 1 false positive; Met: Yes.
Sample Type: QC Control B; Target Conc (log IU/mL): 2.16; Valid N: 40; Expected Result: +-0.5 logs from Value Assignment; Allowable Number of Outliers: 1/30; Standard Deviation:
§ 866.3170 Nucleic acid-based hepatitis C virus ribonucleic acid tests.
(a)
Identification. A nucleic acid-based hepatitis C virus (HCV) ribonucleic acid (RNA) test is identified as an in vitro diagnostic device intended for prescription use as an aid in the diagnosis of HCV infection in specified populations, and/or as an aid in the management of HCV-infected patients including guiding the selection of genotype-specific treatment in individuals with chronic HCV infection. The test is intended for use with human serum or plasma. The test is not intended for use as a donor screening test for the presence of HCV antibodies in blood, blood products, or tissue donors.(b)
Classification. Class II (special controls). The special controls for this device are:(1) For all nucleic acid-based HCV RNA tests, the labeling required under § 809.10(b) of this chapter must include:
(i) A prominent statement that the test is not intended for use as a donor screening test for the presence of HCV RNA from human cells, tissues, and cellular and tissue-based products.
(ii) A detailed explanation of the principles of operation and procedures for performing the assay.
(iii) A detailed explanation of the interpretation of results.
(iv) Limitations, which must be updated to reflect current clinical practice and disease presentation and management. These limitations must include, but are not limited to, statements that indicate:
(A) The specimen types for which the device has been cleared and that use of this test kit with specimen types other than those specifically cleared for this device may result in inaccurate test results.
(B) When applicable, that assay performance characteristics have not been established in populations of immunocompromised or immunosuppressed patients or, other populations where test performance may be affected.
(C) Test results are to be interpreted by qualified licensed healthcare professionals in conjunction with the individual's clinical presentation, history, and other laboratory results.
(2) For all nucleic acid-based HCV RNA tests, the design verification and validation must include:
(i) Detailed device description, including the device components, ancillary reagents required but not provided, and an explanation of the device methodology. Additional information appropriate to the technology must be included such as design of primers and probes, rationale for the selected gene targets, specifications for amplicon size, and degree of nucleic acid sequence conservation.
(ii) For devices with assay calibrators, the design and nature of all primary, secondary, and subsequent quantitation standards used for calibration as well as their traceability to a standardized reference material that FDA has determined is appropriate (
e.g., a recognized consensus standard). In addition, analytical testing must be performed following the release of a new lot of the standard material that was used for device clearance or approval, or when there is a transition to a new calibration standard.(iii) Documentation and characterization (
e.g., determination of the identity, supplier, purity, and stability) of all critical reagents (including nucleic acid sequences for primers and probes) and protocols for maintaining product integrity.(iv) Detailed documentation of analytical performance studies conducted as appropriate to the technology, specimen types tested, and intended use of the device, including, but not limited to, limit of detection (LoD), upper and lower limits of quantitation (ULoQ and LLoQ, respectively), linearity, precision, endogenous and exogenous interferences, cross reactivity, carryover, matrix equivalency, and sample and reagent stability. Samples selected for use in analytical studies or used to prepare samples for use in analytical studies must be from subjects with clinically relevant circulating genotypes in the United States. Cross-reactivity studies must include samples from HCV RNA negative subjects with other causes of liver disease, including autoimmune hepatitis, alcoholic liver disease, chronic hepatitis B virus, primary biliary cirrhosis, and nonalcoholic steatohepatitis, when applicable. The effect of each claimed nucleic-acid isolation and purification procedure on detection must be evaluated.
(v) Risk analysis and management strategies, such as Failure Modes Effects Analysis and/or Hazard Analysis and Critical Control Points summaries and their impact on test performance.
(vi) Final release criteria to be used for manufactured test lots with appropriate evidence that lots released at the extremes of the specifications will meet the claimed analytical and clinical performance characteristics as well as the stability claims.
(vii) Multisite reproducibility study that includes the testing of three independent production lots.
(viii) All stability protocols, including acceptance criteria.
(ix) Final release test results for each lot used in clinical studies.
(x) Analytical sensitivity and specificity of the test must be the same or better than that of other cleared or approved tests.
(xi) Lot-to-lot precision studies, as appropriate.
(3) For devices intended for the qualitative detection of HCV RNA, in addition to the special controls listed in paragraphs (b)(1) and (2) of this section, the design verification and validation must include detailed documentation of performance from a multisite clinical study. Performance must be analyzed relative to an FDA cleared or approved qualitative HCV RNA test, or a comparator that FDA has determined is appropriate. This study must be conducted using appropriate patient samples, with appropriate numbers of HCV positive and negative samples in applicable risk categories. Additional genotypes must be validated using appropriate numbers and types of samples. The samples may be a combination of fresh and repository samples, sourced from within and outside the United States, as appropriate. The study designs, including number of samples tested, must be sufficient to meet the following criteria:
(i) Clinical sensitivity of the test must have a lower bound of the 95 percent confidence interval of greater than or equal to 95 percent.
(ii) Clinical specificity of the test must have a lower bound of the 95 percent confidence interval of greater than or equal to 96 percent.
(4) For devices intended for the quantitative detection of HCV RNA, the following special controls, in addition to those listed in paragraphs (b)(1) and (2) of this section, apply:
(i) Labeling required under § 809.10(b) of this chapter must include a prominent statement that the test is not intended as a diagnostic test to confirm the presence of active HCV infection, when applicable.
(ii) Design verification and validation must include the following:
(A) Detailed documentation of the following analytical performance studies conducted as appropriate to the technology, specimen types tested, and intended use of the device, including but not limited to: LoD, ULoQ and LLoQ. LoD, LLoQ, and linearity studies must demonstrate acceptable device performance with all HCV genotypes detected by the device.
(B) Detailed documentation of clinical performance testing from either:
(
1 ) A multisite clinical study with an appropriate number of clinical samples from chronically HCV infected patients in which the results are compared to an FDA-cleared or approved quantitative HCV RNA test, or a comparator that FDA has determined is appropriate. This study must include a sufficient number of HCV positive samples containing an analyte concentration near the LLoQ to describe performance at this level. Clinical samples must cover the full range of the device output and must be consistent with the distribution of these genotypes in the U.S. population. Clinical samples may be supplemented with diluted clinical samples for those viral load concentrations that are not sufficiently covered by natural clinical specimens, or(
2 ) A clinical study with prospectively collected samples demonstrating clinical validity of the device.(C) Detailed documentation of a qualitative analysis near the lower end of the measuring range demonstrating acceptable performance when used as an aid in diagnosis.
(5) For devices intended for HCV RNA genotyping, in addition to the special controls listed in paragraphs (b)(1) and (2) of this section, design verification and validation must include the following:
(i) Detailed documentation of an analytical performance study demonstrating the LoD for all HCV genotypes detected by the device.
(ii) Detailed documentation, including results, of a multisite clinical study that assesses genotyping accuracy (
i.e., the proportion of interpretable results that match with the reference method result) and the genotyping rate (i.e., the proportion of results that were interpretable).(6) For any nucleic acid-based HCV RNA test intended for Point of Care (PoC) use, the following special controls, in addition to those listed in paragraphs (b)(1) and (2) of this section, apply:
(i) Clinical studies must be conducted at PoC sites.
(ii) Additional labeling must include a brief summary of the instructions for use that are appropriate for use in a PoC environment.
0
July 24, 2024
Image /page/0/Picture/1 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left, there is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.
Hologic, Inc. Howard Liu Regulatory Affairs Specialist 10210 Genetic Center Drive San Diego, California 92121
Re: K233352
Trade/Device Name: Aptima HCV Quant Dx Assay Regulation Number: 21 CFR 866.3170 Regulation Name: Nucleic Acid-Based Hepatitis C Virus Ribonucleic Acid Tests Regulatory Class: Class II Product Code: MZP Dated: June 28, 2024 Received: July 1, 2024
Dear Howard Liu:
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 (the 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 available 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.
Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device" (https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).
1
2
Your device is also subject to, among other requirements, the Quality System (QS) regulation (21 CFR Part 820), which includes, but is not limited to, 21 CFR 820.30, Design controls; 21 CFR 820.90, Nonconforming product; and 21 CFR 820.100, Corrective and preventive action. Please note that regardless of whether a change requires premarket review, the QS regulation requires device manufacturers to review and approve changes to device design and production (21 CFR 820.30 and 21 CFR 820.70) and document changes and approvals in the device master record (21 CFR 820.181).
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR Part 803) for devices or postmarketing safety reporting (21 CFR Part 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safetyreporting-combination-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 Part 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR Parts 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 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,
Maria I. Garcia -S
Maria Garcia, Ph.D. Assistant Director Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health
2
Indications for Use
510(k) Number (if known) K233352
Device Name Aptima HCV Quant Dx Assay
Indications for Use (Describe)
The Aptima HCV Quant Dx assay is a real-time transcription-mediated amplification (TMA) test used for both detection and quantitation of hepatitis C virus (HCV) RNA in fresh and frozen human serum and plasma from HCV-infected individuals.
Plasma may be prepared in ethylenediaminetetraacetic acid (EDTA), anticoagulant citrate dextrose (ACD) solution, and plasma preparation tubes (PPT). Serum may be prepared in serum separator tubes (SST). Specimens are tested using the Panther system for automated specimen processing, and quantitation. Specimens containing HCV genotypes 1 to 6 are validated for detection and quantitation in the assay.
The Aptima HCV Quant Dx assay is indicated for use as an aid in the diagnosis of active HCV infection in the following populations: individuals with antibody evidence of HCV infection with evidence of liver disease of to be actively infected with HCV antibody evidence, and individuals at risk for HCV infection with antibodies to HCV. Detection of HCV RNA indicates that the virus is replicating and, therefore, is evidence of active infection of HCV RNA does not discriminate between acute and chronic states of infection.
The Aptima HCV Quant Dx assay is also indicated for use as an aid in the management of HCV infected patients undergoing HCV antiviral drug therapy. The assay can be used to measure HCV RNA levels periodically prior to, during, and after treatment to determine sustained virological response (SVR) or nonsustained virological response (NSVR), Assay performance characteristics have been established for individuals infected with HCV and treated with certain direct-acting antiviral agents (DAA) regimens. No information is available on the assay's predictive value when other therapies are used. The results from the Aptima HCV Quant Dx assay must be interpreted within the context of all relevant clinical and laboratory findings.
The Aptima HCV Quant Dx assay is not approved for use as a screening test for the presence of HCV RNA in blood or blood products.
| Type of Use (Select one or both, as applicable) |
|---|
| ------------------------------------------------- |
| ☒ Exportation (Dual-Use, 991 Schedule B) | ☐ Service/Consulting (NLR, 991 Schedule B) |
|---|---|
| ------------------------------------------------------------------------------------------ | -------------------------------------------- |
|X | Prescription Use (Part 21 CFR 801 Subpart D)
| | Over-The-Counter Use (21 CFR 801 Subpart C)
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3
HOLO
510(k) SUMMARY Aptima HCV Quant Dx Assay
I. SUBMITTER
Hologic, Inc. 10210 Genetic Center Drive San Diego, CA 92121
- Contact Person: Howard Liu Regulatory Affairs Specialist howard.liu@hologic.com Phone: (858) 410-8853
Date Prepared: July 18, 2024
II. DEVICE
Aptima HCV Quant Dx assay Proprietary Name of Device: Regulation Description: Nucleic acid-based hepatitis C virus ribonucleic acid tests Classification Panel/Medical Specialty: Microbiology Regulation Number: 21 CFR 866.3170 Regulatory Class: Class II Product Code: MZP
III. PREDICATE DEVICE
The predicate device is the Aptima HCV Quant Dx assay; P160023, originally approved as a
Class III PMA product, reclassified to a Class II 510(k) product on November 26, 2021.
4
IV. DEVICE DESCRIPTION
Clearance of this pre-market application will 1) enable additional reagent on-board / off-board cycling, allowing operators to load reagents onto the Panther System 8 times instead of 5 times; and 2) update the Assay Definition Module (ADM) to detect, flag and invalidate results impacted by a faulty or flickering LED, using estimated background minimum limits. These changes do not introduce any changes to the original design, method of manufacture, assay procedure, principle of operation, mechanism of action, conditions of use or hardware of the Panther instrument, or to the results interpretation for the cleared assay.
Principles of the Procedure
The principles of the procedure are unchanged.
The Aptima HCV Quant Dx assay is a nucleic acid amplification test that uses real-time TMA technology to detect and quantitate HCV RNA for aiding diagnosis or to establish baseline viral load, as well as to measure on-treatment and post-treatment responses. The assay targets a conserved region of the HCV genome, detecting and quantitating genotypes 1, 2, 3, 4, 5, and 6. The assay is standardized against the 2nd WHO International Standard for Hepatitis C Virus (NIBSC Code 96/798).12
The Aptima HCV Quant Dx assay involves three main steps, which all take place in a single tube on the Panther system: target capture, target amplification by TMA, and detection of the amplification products (amplicon) by the fluorescent labeled probes (torches).
During target capture, viral RNA is isolated from specimens. The specimen is treated with a detergent to solubilize the viral envelope, denature proteins, and release viral genomic RNA. Capture oligonucleotides hybridize to highly conserved regions of HCV RNA, if present, in the test specimen. The hybridized target is then captured onto magnetic microparticles that are separated from the specimen in a magnetic field. Wash steps remove extraneous components from the reaction tube.
5
Target amplification occurs via TMA, which is a transcription-mediated nucleic acid amplification method that utilizes two enzymes, Moloney murine leukemia virus (MMLV) reverse transcriptase and T7 RNA polymerase. The reverse transcriptase is used to generate a DNA copy (containing a promoter sequence for T7 RNA polymerase) of the target sequence. T7 RNA polymerase produces multiple copies of RNA amplicon from the DNA copy template. The Aptima HCV Quant Dx assay utilizes the TMA method to amplify a portion of the 5' UTR of the HCV genome. Amplification of this region is achieved using specific primers which are designed to amplify HCV genotypes 1, 2, 3, 4, 5, and 6.
Detection is achieved using single-stranded nucleic acid torches that are present during the amplification of the target and that hybridize specifically to the amplicon in real time. Each torch has a fluorophore and a quencher. When the torch is not hybridized to the amplicon, the quencher is in close proximity of the fluorophore and suppresses the fluorescence. When the torch binds to the amplicon, the quencher is moved farther away from the fluorophore and it will emit a signal at a specific wavelength when excited by a light source. As more torches hybridize to amplicon a higher fluorescent signal is generated. The time taken for the fluorescent signal to reach a specified threshold is proportional to the starting HCV concentration. Each reaction has an internal calibrator/internal control (IC) that controls for variations in specimen processing, amplification, and detection. The concentration of a sample is determined by the Panther system software using the HCV and IC signals for each reaction and comparing them to calibration information.
Assay Components
The reagents required to perform the Aptima HCV Quant Dx assay are available in 100-test kits. The kits are packaged in 3 boxes containing 11 reagents which are required for sample processing. A description of the components that are required to perform the Aptima HCV Quant Dx assay are detailed in Table 1. In addition, two ancillary kits are required to run the assay (Table 2).
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| Box | Components Description |
|---|---|
| 1 | Amplification Reagent |
| Enzyme Reagent | |
| Promoter Reagent | |
| Amplification Reconstitution Solution | |
| Enzyme Reconstitution Solution | |
| Promoter Reconstitution Solution | |
| Target Capture Reagent | |
| 2 | Positive Calibrator |
| Negative Control | |
| 3 | Low Positive Control |
| High Positive Control |
Table 1: Reagents Required to Perform the Aptima HCV Quant Dx Assay
| Table 2: Ancillary Kits Required to Perform the Aptima HCV Quant Dx Assav | |||||
|---|---|---|---|---|---|
| Panther Run Kit for Real Time Assays (for real time assays only) |
|---|
| Panther System Run Kit |
| (when running non-real time-TMA assays in parallel with real |
| time-TMA assays) |
Instrumentation
The Aptima HCV Quant Dx assay has been designed for and validated on the Panther system. The Panther system is an integrated hardware and software system that together with the Aptima HCV Quant Dx assay fully automates all the steps necessary to perform the assay from sample preparation through amplification of nucleic acid, detection, data reduction and amplicon inactivation.
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V. INDICATIONS FOR USE
Intended Use
The intended use for the Aptima HCV Quant Dx Assay will remain unchanged:
The Aptima HCV Quant Dx assay is a real-time transcription-mediated amplification (TMA) test used for both detection and quantitation of hepatitis C virus (HCV) RNA in fresh and frozen human serum and plasma from HCV-infected individuals.
Plasma may be prepared in ethylenediaminetetraacetic acid (EDTA), anticoagulant citrate dextrose (ACD) solution, and plasma preparation tubes (PPT). Serum may be prepared in serum tubes and serum separator tubes (SST). Specimens are tested using the Panther® system for automated specimen processing, amplification, detection, and quantitation. Specimens containing HCV genotypes 1 to 6 are validated for detection and quantitation in the assay.
The Aptima HCV Quant Dx assay is indicated for use as an aid in the diagnosis of active HCV infection in the following populations: individuals with antibody evidence of HCV infection with evidence of liver disease, individuals suspected to be actively infected with HCV antibody evidence, and individuals at risk for HCV infection with antibodies to HCV. Detection of HCV RNA indicates that the virus is replicating and, therefore, is evidence of active infection. Detection of HCV RNA does not discriminate between acute and chronic states of infection.
The Aptima HCV Quant Dx assay is also indicated for use as an aid in the management of HCV infected patients undergoing HCV antiviral drug therapy. The assay can be used to measure HCV RNA levels periodically prior to, during, and after treatment to determine sustained virological response (SVR) or nonsustained virological response (NSVR). Assay performance characteristics have been established for individuals infected with HCV and treated with certain direct-acting antiviral agents (DAA) regimens. No information is available on the assay's predictive value when other therapies are used. The results from the Aptima HCV Quant Dx assay must be interpreted within the context of all relevant clinical and laboratory findings.
The Aptima HCV Quant Dx assay is not approved for use as a screening test for the presence of HCV RNA in blood or blood products.
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VI. COMPARISON OF TECHNOLOGICAL CHARACTERISTICS WITH THE PREDICATE DEVICE
A comparison of the Aptima HCV Quant Dx assay with the updated 7.2.9 system software to the predicate device, the Aptima HCV Quant Dx assay (P160023, originally approved as a Class III PMA product, reclassified to a Class II 510(k) product on November 26, 2021), is summarized in Table 3 (similarities) and Table 4 (differences).
| Item | Predicate Device:
Aptima HCV Quant Dx assay
(P160023) | Subject Device:
Aptima HCV Quant Dx assay |
|--------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------|
| Technology Principle
of Operation | Target Capture (TC), Transcription-
Mediated Amplification (TMA) | Same |
| Platform | Automated Panther System | Same |
| Assay Targets | Hepatitis C RNA | Same |
| Assay Results | Qualitative & quantitative | Same |
| Function | Detection of RNA from Hepatitis C | Same |
| Intended Use | The Aptima HCV Quant Dx assay is a real-time
transcription-mediated amplification (TMA)
test used for both detection and quantitation of
hepatitis C virus (HCV) RNA in fresh and
frozen human serum and plasma from HCV-
infected individuals.
Plasma may be prepared in
ethylenediaminetetraacetic acid (EDTA),
anticoagulant citrate dextrose (ACD) solution,
and plasma preparation tubes (PPT). Serum
may be prepared in serum tubes and serum
separator tubes (SST). Specimens are tested
using the Panther® system for automated
specimen processing, amplification, detection,
and quantitation. Specimens containing HCV
genotypes 1 to 6 are validated for detection and
quantitation in the assay.
The Aptima HCV Quant Dx assay is indicated
for use as an aid in the diagnosis of active HCV
infection in the following populations:
individuals with antibody evidence of HCV
infection with evidence of liver disease,
individuals suspected to be actively infected
with HCV antibody evidence, and individuals | Same |
| Item | Predicate Device:
Aptima HCV Quant Dx assay
(P160023) | Subject Device:
Aptima HCV Quant Dx assay |
| | HCV. Detection of HCV RNA indicates that
the virus is replicating and, therefore, is
evidence of active infection. Detection of HCV
RNA does not discriminate between acute and
chronic states of infection. | |
| | The Aptima HCV Quant Dx assay is also
indicated for use as an aid in the management
of HCV infected patients undergoing HCV
antiviral drug therapy. The assay can be used to
measure HCV RNA levels periodically prior to,
during, and after treatment to determine
sustained virological response (SVR) or
nonsustained virological response (NSVR).
Assay performance characteristics have been
established for individuals infected with HCV
and treated with certain direct-acting antiviral
agents (DAA) regimens. No information is
available on the assay's predictive value when
other therapies are used. The results from the
Aptima HCV Quant Dx assay must be
interpreted within the context of all relevant
clinical and laboratory findings. | |
| | The Aptima HCV Quant Dx assay is not
approved for use as a screening test for the
presence of HCV RNA in blood or blood
products. | |
Table 3: Similarities Between Predicate Device and Subject Device
K233352-S002.510kSummary.FINAL.docx Original 510(k) Application
9
Table 4: Differences Between Predicate Device and Subject Device
| Item | Predicate Device:
Aptima HCV Quant Dx assay
(P160023) | Subject Device:
Aptima HCV Quant Dx assay |
|-----------------------------------------------------------------------------|-------------------------------------------------------------|----------------------------------------------|
| Reagents can be
loaded onto the
Panther system | 5 times | 8 times |
| Results impacted by
faulty or flickering
LED in Panther
instrument | Read as normal,
potentially producing incorrect results | Marked as invalid results |
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VII. PERFORMANCE DATA
Data provided in support of the substantial equivalence determination consist of the following validation activities:
- Intended Use Testing
- Assay Verification ●
- . Software Verification and Validation
Intended Use Testing
Results from the Aptima HCV Quant Assay on Panther Fusion System System Software 7.2.9 were compared to those from the previous software version, 7.2.7 (reported to the FDA in the 2022 PMA annual report for the Aptima HCV Quant Dx Assay, P160023/R004).
The results met established acceptance criteria, confirming that assay performance is equivalent to the prior comparators, and demonstrating that the system is capable of performing the Aptima HCV Quant Dx Assay version 5.3.5.1 on the Panther Fusion System Software 7.2.9.
| Sample
Type | Target
Conc
(log
IU/mL)*** | Valid
N | Expected
Result | Allowable
Number
of
Outliers | Standard
Deviation* | Result | Met? |
|--------------------|-------------------------------------|------------|-----------------------------------------|---------------------------------------|------------------------|-------------------------------------------------------------------------------------------------------------------------|------|
| QC
Control
A | Negative | 120 | Not Detected | 1 False
positive**** | N/A | 1 false positive | Met |
| QC
Control
B | 2.16 | 40 | ±0.5 logs
from Value
Assignment** | 1/30 | ≤ 0.25 log
copy* | 100% Positivity
Ave diff between expected
and recovered is -0.05 log
IU/mL and largest SD is 0.07
log IU/mL | Met |
| QC
Control
D | 6.52 | 40 | ±0.5 logs
from Value
Assignment** | 0/25 | ≤ 0.25 log
copy | 100% Positivity
Ave diff between expected
and recovered is -0.12 log
IU/mL and largest SD is 0.03
log IU/mL | Met |
| QC
Panel S | 1.13 | 80 | 95%
Positivity | N/A | N/A | 100% Positivity | Met |
may be removed from calculation of Standard Deviatio logs from Target Concentration if Value Assignm
** ±0.50 logs from Target Concentration if Value
*** Using conversion factor of 4,4 HCV copies/IU
ecificity with 95% confidence int
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Assay Verification (5 to 8 reagent reload change)
Aptima HIV-1 Quant, Aptima HCV Quant Dx and Aptima HBV Quant Assay reagents were subjected to at least 72 cumulative hours on the Panther system with the reagents uncapped. Testing included 5-10 cycles of reagent storage between the Panther system and the assigned reagent storage temperature of 2-8°C. Assay testing was performed on Day 0 and after cycling. Five cycles are the current maximum of cycles allowed for each assay. Testing included at least one reagent lot of each assay stored on-board 1 Panther system at the environmental extremes for reagent evaporation loss (30°C and 20% relative humidity). Performance was evaluated using prepared virus and armored RNA (aRNA) spiked into negative human serum matrix (BI0052).
Aptima HBV and HCV reagents successfully completed 10 on and off on-board cycles with a cumulative 72 hours on board stability.
| HCV 3.3 log IU/mL | |||||
|---|---|---|---|---|---|
| Kit Cycles | Total Hours | Cycle Avg LogIU/mL | Cycle Std Dev | Baseline Avg LogIU/mL | Diff (Cycle-Baseline) |
| 5 | 35 | 3.22 | 0.05 | 3.20 | 0.02 |
| 10 | 72 | 3.28 | 0.06 | 3.10 | 0.18 |
| HCV 7.0 log IU/mL | |||||
|---|---|---|---|---|---|
| Kit Cycles | Total Hours | Cycle Avg LogIU/mL | Cycle Std Dev | Baseline Avg LogIU/mL | Diff (Cycle-Baseline) |
| 5 | 35 | 7.45 | 0.15 | 7.56 | -0.11 |
| 10 | 72 | 7.30 | 0.07 | 7.56 | -0.26 |
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Assay Verification (Flickering LED process control change)
Fluorescent curve files and logs from 381 field instruments and 445,910 unique test orders from March 2018 to March 2022 were analyzed. Test orders with outlier estimated background values were excluded by specific root-cause analysis where applicable. Incomplete fluorescent curve files due to a date split were excluded from analysis.
HCV Estimated Background distributions were analyzed by each channel, FAM and ROX as seen below. Data were approximately normal for each channel. Proposed estimated background limits would be 50 or five standard deviations from the mean, respectively. One test order was flagged by FAM estimated background limit and was also flagged by QNS; this test order was considered a true fault.
Image /page/12/Figure/5 description: The image shows two histograms and summary statistics for HCV Estimated Background (FAM) and HCV Estimated Background (ROX). For FAM, the mean is 988, the standard deviation is 151, and N is 120,878. For ROX, the mean is 1,270, the standard deviation is 245, and N is 120,862. Each histogram shows a distribution of values, with the FAM distribution centered around 1000 and the ROX distribution centered around 1200.
This analysis supports the implementation of the proposed Estimated Background minimum limits as seen in Table 1 for HIV, HCV, HBV, and HSV Panther viral assays. New estimated background limits will increase fault detection for these assays.
| FAM | HEX | ROX | |
|---|---|---|---|
| HIV | 200 | 1,000 | 150 |
| HCV | 300 | N/A | 100 |
| HBV | 300 | 200 | 100 |
| HSV | 300 | 150 | 100 |
K233352-S002.510kSummary.FINAL.docx Original 510(k) Application
Confidential
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Software Verification and Validation
SW V&V testing was performed to ensure that the Panther Fusion System Software, Version 7.2.9 conforms to user needs and intended uses, and that the requirements implemented through software are consistently fulfilled. Software V&V testing of applicable software-specific Product Requirements Document (PRD), Software Requirement Specifications (SRS), and Product Specification Document (PSD) requirements, including regression testing, as applicable, were performed, with the results summarized below. All anomalies generated as part of this testing were reviewed in Defect Review Board (DRB) meetings in accordance with the Software Anomaly Management Work Instruction, 04-03-21-001-WI.
| Requirements | Pass | Fail |
|---|---|---|
| PRM Requirements | 23 | 0 |
| PRD Requirements | 29 | 0 |
| SRS Requirements | 210 | 0 |
| PSD Requirements | 4 | 0 |
| Regression testing of Anomalies Addressed and | ||
| Implemented Features and Change Requests | 12 | 0 |
The reporting and assessment of anomalies was conducted in accordance with the Software Anomaly Management Work Instruction, 04-03-21-001-WI and Product Safety Risk Management Procedure, 04-03-12-SOP. Each software anomaly was assessed for impact to patient and user safety. The assessment determined there are no known software anomalies that exceed the severity level of "Negligible". The following table summarizes the total number of identified anomalies associated with Panther and Panther Fusion System Software, Version 7.2.9.
| Severity Level | Definition | Total |
|---|---|---|
| Catastrophic | Directly results in patient or user death | 0 |
| Critical | Directly results in irreversible injury (permanent impairment or | |
| life-threatening injury) | 0 | |
| Serious | Results in reversible injury or impairment requiring professional | |
| medical intervention | 0 | |
| Minor | Results in temporary injury or impairment not requiring professional | |
| medical intervention or blood supply delay or treatment delay | 0 | |
| Negligible | Results in discomfort/no impact | 12 |
It has been concluded from this testing that the Panther Fusion System Software provides safe and effective performance with the essential features for the end user to meet the intended use.
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SW V&V testing was also performed for the Panther Aptima HCV ADM on Panther System Software Version 7.2.9. As above, system Validation activities included testing of applicable requirements or specifications from the Product Requirements Document (PRD), Product Risk Mitigator (PRM), Product Specification Document (PSD) and Software Requirement Specification Document (SRS) as well as testing of modifications associated with hardware, anomaly fixes and/or Software Change Requests (SCR).
There were no anomalies identified during the V&V of the Panther Aptima HCV ADM on Panther System Software Version 7.2.9.
It has been concluded from the overall testing that the Panther Aptima HCV assay on the Panther System Software Version 7.2.9 meets the intended use and safety requirements.
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Additional Studies
The FDA issued a hold letter on January 3, 2024 with a request for additional information in two parts as described below, with Hologic responses following:
1. Intended Use Testing
- An Intended Use Testing study using a minimum of four (4) clinical specimens including: . three (3) specimens/samples with HCV RNA concentration around 3xLLoQ (30 IU/ml) and one (1) HCV RNA negative clinical specimen. Samples may be prepared by diluting a high positive sample in negative clinical matrix. FDA recommends testing 10 replicates of each sample per run, using previous Aptima HCV software version 5.2.5.1 and the updated version 5.3.5.1.
- A revised study report including a clear description of the study and the nature of samples . used (e.g., natural or contrived), data analysis, summary of the results and conclusions, and line data in Excel format.
The requested study was carried out, Aptima HCV Assay Software versions 5.2.5.1 and 5.3.5.1 successfully completed and met all acceptance criteria with low level clinical panel. The Aptima HCV Assay Software version 5.2.5.1 and Aptima HCV Assay Software version 5.3.5.1 both resulted in 100% not detected for the negative clinical matrix panel. The Aptima HCV Assay Software version 5.2.5.1 and Aptima HCV Assay Software version 5.3.5.1 both resulted in 100% positivity for the 30 IU/mL (low positive) clinical positive panel. Acceptance criteria for this panel is positivity only and has no accuracy or precision specifications. The requested files are included in this supplement.
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2. On-Board Stability (OBS) and Reagent Cycling
To support the additional reagent on-board/off-board cycling, please provide the following:
- . An OBS and reagent cycling study including a minimum of one negative sample, a sample around 3xLLoQ: 30 IU/ml, and a moderate positive or high positive sample. Please include a minimum of 5 replicates per sample for each timepoint with more replicates tested at baseline to establish a robust baseline. Please perform the testing using the updated Panther system software version 7.2.9 and Aptima HCV software version 5.3.5.1 or provide a justification for why software changes would not impact the reagent stability results.
- A revised study report, including a detailed study description and the nature of samples . used (e.g., natural or contrived), data analysis, summary of the results, line data in Excel format, and conclusions clearly stating the supported OBS and reagent cycling claim. Note, in order to support a claim of 8 times reagent cycling, please evaluate at minimum, the number of cycles previously claimed (5 cycles), the number of cycles currently claimed (8 cycles) and an additional number of cycles that goes beyond the one currently claimed (9 to 10 cycles) using the same kit when possible. This ensures that the reagent stability does not change one time point after the desired claim.
The requested study was carried out, and Aptima reagents successfully completed five, eight, and ten on and off on-board cycles with a cumulative 72 hours on board stability and met all acceptance criteria.
VIII. CONCLUSIONS
The validation study results demonstrate that the Aptima HCV Quant Dx assay on the 7.2.9 Panther system software performs comparably to the predicate device and supports a substantial equivalence decision.