(105 days)
The Aptima® SARS-CoV-2 Assay is a nucleic acid amplification in vitro diagnostic test intended for the qualitative detection of RNA from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) isolated and purified from nasopharyngeal (NP) swab and anterior nasal (AN) swab specimens obtained from patients with signs and symptoms of COVID-19.
Positive results are indicative of the presence of SARS-CoV-2 RNA. The Aptima SARS-CoV-2 Assay is intended for use as an aid in the diagnosis of COVID-19 if used in conjunction with other clinical, epidemiological, and laboratory findings. Clinical correlation with patient history and other diagnostic information is necessary to determine patient infection status. Positive results do not rule out bacterial infection or co-infection with other viruses.
Negative results do not preclude SARS-CoV-2 infection and should not be used as the sole basis for patient management decisions. Negative results must be combined with clinical observations, patient history, and epidemiological information.
The Aptima SARS-CoV-2 Assay is a nucleic acid amplification in vitro diagnostic test developed for use on the fully automated Panther/Panther Fusion system to detect RNA from SARS-CoV-2 isolated and purified from nasopharyngeal and anterior nasal swab specimens collected into UTM/VTM or with the RespDirect Collection Kit.
The Aptima SARS-CoV-2 Assay combines the technologies of target capture, Transcription Mediated Amplification (TMA), and Dual Kinetic Assay (DKA).
Specimens are collected and transferred into their respective specimen transport tubes. The transport solutions in these tubes release the RNA target and protect them from degradation during storage. When the Aptima SARS-CoV-2 Assay is performed in the laboratory, the target RNA molecules are isolated from specimens by use of capture oligomers via target capture that utilizes magnetic microparticles. The capture oligomers contain sequences complementary to specific regions of the target molecules as well as a string of deoxyadenosine residues. A separate capture oligomer is used for each target. During the hybridization step, the sequence specific regions of the capture oligomers bind to specific regions of the target molecules. The capture oligomer:target complex is then captured out of solution by decreasing the temperature of the reaction to room temperature. This temperature reduction allows hybridization to occur between the deoxyadenosine region on the capture oligomer and the poly-deoxythymidine molecules that are covalently attached to the magnetic particles. The microparticles, including the captured target molecules bound to them, are pulled to the side of the reaction vessel using magnets and the supernatant is aspirated. The particles are washed to remove residual specimen matrix that may contain amplification reaction inhibitors. After the target capture steps are completed, the specimens are ready for amplification.
Target amplification assays are based on the ability of complementary oligonucleotide primers to specifically anneal and allow enzymatic amplification of the target nucleic acid strands. The Aptima SARS-CoV-2 Assay replicates specific regions of the RNA from SARS-CoV-2 virus. Detection of the RNA amplification product sequences (amplicon) is achieved using nucleic acid hybridization. Single-stranded chemiluminescent nucleic acid probes, which are unique and complementary to a region of each target amplicon and Internal Control (IC) amplicon, are labeled with different acridinium ester (AE) molecules. The AE-labeled probes combine with the amplicon to form stable hybrids. The Selection Reagent differentiates hybridized from unhybridized probe, eliminating the generation of signal from the unhybridized probe. During the detection step, light emitted from the labeled hybrids is measured as photon signals in a luminometer and are reported as Relative Light Units (RLU). In DKA, differences in the kinetic profiles of the labeled probes allow for the differentiation of signal; kinetic profiles are derived from measurements of photon output during the detection read time.
The Aptima SARS-CoV-2 Assay amplifies and detects 2 conserved regions of the ORF1ab gene in the same reaction, using the "glower" kinetic type. The 2 regions are not differentiated and amplification of either or both regions lead to RLU signal. The assay results are determined by a cut-off based on the total RLU and the kinetic curve type.
Here's a breakdown of the acceptance criteria and the study that proves the device meets them, based on the provided text:
Acceptance Criteria and Reported Device Performance
The acceptance criteria are implicitly defined by the performance metrics presented as evidence of substantial equivalence to the predicate device. The key metrics are Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA). While explicit numerical acceptance thresholds are not provided in the document, the reported performance demonstrates "comparable" results to the predicate device.
| Performance Metric | Acceptance Criteria (Implicit) | Reported Device Performance (NP Swab, Overall) | Reported Device Performance (AN Swab, UTM/VTM) | Reported Device Performance (AN Swab, RespDirect eSTM) |
|---|---|---|---|---|
| Clinical Performance: | ||||
| Positive Percent Agreement (PPA) | High Agreement | 95.4% (90.3-97.9% CI) | 96.5% (92.1-98.5% CI) | 100% (96.6-100% CI) |
| Negative Percent Agreement (NPA) | High Agreement | 99.5% (98.9-99.8% CI) | 97.6% (96.5-98.4% CI) | 98.0% (96.9-98.7% CI) |
| Analytical Performance (Key): | ||||
| Limit of Detection (LoD) - Cultured Virus | Extremely Low Concentration | 0.01 TCID50/mL | Not applicable (analytical, not specimen-type specific) | Not applicable (analytical, not specimen-type specific) |
| Limit of Detection (LoD) - WHO Standard | Extremely Low Concentration | 87.5 IU/mL | Not applicable (analytical, not specimen-type specific) | Not applicable (analytical, not specimen-type specific) |
| Reactivity (detection of strains) | 100% Positivity at low concentrations | Demonstrated at 0.03-0.151 TCID50/mL for tested strains; 99.98% by in silico analysis | Not applicable (analytical, not specimen-type specific) | Not applicable (analytical, not specimen-type specific) |
| Specificity/Microbial Interference | No cross-reactivity/interference | No observed cross-reactivity/interference for 48 tested organisms | Not applicable (analytical, not specimen-type specific) | Not applicable (analytical, not specimen-type specific) |
| Interfering Substances | No impact on performance | No impact on performance for tested substances | Not applicable (analytical, not specimen-type specific) | Not applicable (analytical, not specimen-type specific) |
| Carryover Contamination Rate | Low/None | 0% (0/294) | Not applicable (analytical, not specimen-type specific) | Not applicable (analytical, not specimen-type specific) |
| Assay Precision (Total %CV of signal) | Low variability | 2.75% to 3.84% (Negative, Low, Moderate Positive panels) | Not applicable (analytical, not specimen-type specific) | Not applicable (analytical, not specimen-type specific) |
Study Details:
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Sample Sizes Used for the Test Set and Data Provenance:
- Clinical Study 1 (NP Swab Specimens):
- Total Enrolled: 1646 specimens
- Evaluable (Final Data Set): 1495 NP swab specimens (1195 fresh, 300 frozen)
- Provenance: Prospective multicenter study from four participating US pediatric/adolescent, private and/or university hospitals. Specimens collected between June-July 2020 and January-April 2023. Remnant specimens.
- Clinical Study 2 (Anterior Nasal Swab Specimens):
- Total Enrolled Subjects: 2301
- Evaluable Subjects: 2177 individuals (1159 with evaluable anterior nasal swab specimens in UTM/VTM, and 1018 with evaluable nasal swab specimens in eSTM).
- Provenance: Prospective, multicenter clinical study at nine geographically and ethnically diverse US sites during the 2022-2023 respiratory season.
- Clinical Study 1 (NP Swab Specimens):
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Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications:
- The document does not specify the number or qualifications of experts directly establishing the clinical ground truth. Instead, it relies on a Composite Comparator Algorithm (CCA). The CCA consists of "two highly sensitive US FDA EUA SARS-CoV-2 molecular tests" and a "validated PCR followed by bi-directional sequencing (PCR/BDS) assay." This implies that the 'expertise' comes from the validation and regulatory clearance of these comparator assays, rather than individual human experts adjudicating each case.
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Adjudication Method for the Test Set:
- Composite Comparator Algorithm (CCA): "A final CCA result was assigned when two of the three comparator assay results were in concordance." This serves as the adjudication method for determining the true positive/negative status of the clinical samples.
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If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done:
- No. This type of study (MRMC) is typically performed for diagnostic imaging devices where human readers interpret images with and without AI assistance. The Aptima SARS-CoV-2 Assay is an in vitro diagnostic (IVD), a laboratory test that detects nucleic acids, not an imaging device requiring human interpretation of visual data. Therefore, an MRMC study is not applicable here.
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If a Standalone (i.e., algorithm only without human-in-the-loop performance) Was Done:
- Yes, this is effectively a standalone performance study. The Aptima SARS-CoV-2 Assay is an automated nucleic acid amplification test run on the Panther/Panther Fusion system. Its performance is directly compared to the CCA, without human intervention in the interpretation of the device's results.
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The Type of Ground Truth Used:
- Clinical Studies: Composite Comparator Algorithm (CCA) based on the concordance of results from two highly sensitive FDA EUA SARS-CoV-2 molecular tests and a validated PCR/BDS assay. This is a form of reference standard derived from established and highly sensitive laboratory methods.
- Analytical Studies (e.g., LoD, Reactivity, Specificity): The ground truth was established by known concentrations of SARS-CoV-2 virus strains or other microorganisms/interfering substances, prepared in controlled laboratory settings (e.g., "spiked with inactivated cultured SARS-CoV-2 virus").
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The Sample Size for the Training Set:
- The document describes performance evaluation studies (analytical and clinical) for market clearance. It does not provide information on the specific training set size used for the development or training of the assay's internal algorithms (e.g., for the kinetic curve analysis or cut-off determination). This information would typically be part of the assay development and validation, not necessarily detailed in a 510(k) summary unless it significantly changed or impacted performance during the clearance process for the specific assay rather than the underlying platform. The focus here is on the performance of the final, already "trained" device.
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How the Ground Truth for the Training Set Was Established:
- As noted above, details regarding the training set's ground truth establishment are not provided in this 510(k) summary. For IVD devices, ground truth for training internal algorithms typically involves using characterized positive and negative clinical samples, spiked samples with known viral loads, and potentially synthetic data, all carefully confirmed by highly sensitive reference methods or gold standard assays during the R&D phase of the product.
§ 866.3981 Device to detect and identify nucleic acid targets in respiratory specimens from microbial agents that cause the SARS-CoV-2 respiratory infection and other microbial agents when in a multi-target test.
(a)
Identification. A device to detect and identify nucleic acid targets in respiratory specimens from microbial agents that cause the SARS-CoV-2 respiratory infection and other microbial agents when in a multi-target test is an in vitro diagnostic device intended for the detection and identification of SARS-CoV-2 and other microbial agents when in a multi-target test in human clinical respiratory specimens from patients suspected of respiratory infection who are at risk for exposure or who may have been exposed to these agents. The device is intended to aid in the diagnosis of respiratory infection in conjunction with other clinical, epidemiologic, and laboratory data or other risk factors.(b)
Classification. Class II (special controls). The special controls for this device are:(1) The intended use in the labeling required under § 809.10 of this chapter must include a description of the following: Analytes and targets the device detects and identifies, the specimen types tested, the results provided to the user, the clinical indications for which the test is to be used, the specific intended population(s), the intended use locations including testing location(s) where the device is to be used (if applicable), and other conditions of use as appropriate.
(2) Any sample collection device used must be FDA-cleared, -approved, or -classified as 510(k) exempt (standalone or as part of a test system) for the collection of specimen types claimed by this device; alternatively, the sample collection device must be cleared in a premarket submission as a part of this device.
(3) The labeling required under § 809.10(b) of this chapter must include:
(i) A detailed device description, including reagents, instruments, ancillary materials, all control elements, and a detailed explanation of the methodology, including all pre-analytical methods for processing of specimens;
(ii) Detailed descriptions of the performance characteristics of the device for each specimen type claimed in the intended use based on analytical studies including the following, as applicable: Limit of Detection, inclusivity, cross-reactivity, interfering substances, competitive inhibition, carryover/cross contamination, specimen stability, precision, reproducibility, and clinical studies;
(iii) Detailed descriptions of the test procedure(s), the interpretation of test results for clinical specimens, and acceptance criteria for any quality control testing;
(iv) A warning statement that viral culture should not be attempted in cases of positive results for SARS-CoV-2 and/or any similar microbial agents unless a facility with an appropriate level of laboratory biosafety (
e.g., BSL 3 and BSL 3+, etc.) is available to receive and culture specimens; and(v) A prominent statement that device performance has not been established for specimens collected from individuals not identified in the intended use population (
e.g., when applicable, that device performance has not been established in individuals without signs or symptoms of respiratory infection).(vi) Limiting statements that indicate that:
(A) A negative test result does not preclude the possibility of infection;
(B) The test results should be interpreted in conjunction with other clinical and laboratory data available to the clinician;
(C) There is a risk of incorrect results due to the presence of nucleic acid sequence variants in the targeted pathogens;
(D) That positive and negative predictive values are highly dependent on prevalence;
(E) Accurate results are dependent on adequate specimen collection, transport, storage, and processing. Failure to observe proper procedures in any one of these steps can lead to incorrect results; and
(F) When applicable (
e.g., recommended by the Centers for Disease Control and Prevention, by current well-accepted clinical guidelines, or by published peer-reviewed literature), that the clinical performance may be affected by testing a specific clinical subpopulation or for a specific claimed specimen type.(4) Design verification and validation must include:
(i) Detailed documentation, including performance results, from a clinical study that includes prospective (sequential) samples for each claimed specimen type and, as appropriate, additional characterized clinical samples. The clinical study must be performed on a study population consistent with the intended use population and compare the device performance to results obtained using a comparator that FDA has determined is appropriate. Detailed documentation must include the clinical study protocol (including a predefined statistical analysis plan), study report, testing results, and results of all statistical analyses.
(ii) Risk analysis and documentation demonstrating how risk control measures are implemented to address device system hazards, such as Failure Modes Effects Analysis and/or Hazard Analysis. This documentation must include a detailed description of a protocol (including all procedures and methods) for the continuous monitoring, identification, and handling of genetic mutations and/or novel respiratory pathogen isolates or strains (
e.g., regular review of published literature and periodic in silico analysis of target sequences to detect possible mismatches). All results of this protocol, including any findings, must be documented and must include any additional data analysis that is requested by FDA in response to any performance concerns identified under this section or identified by FDA during routine evaluation. Additionally, if requested by FDA, these evaluations must be submitted to FDA for FDA review within 48 hours of the request. Results that are reasonably interpreted to support the conclusion that novel respiratory pathogen strains or isolates impact the stated expected performance of the device must be sent to FDA immediately.(iii) A detailed description of the identity, phylogenetic relationship, and other recognized characterization of the respiratory pathogen(s) that the device is designed to detect. In addition, detailed documentation describing how to interpret the device results and other measures that might be needed for a laboratory diagnosis of respiratory infection.
(iv) A detailed device description, including device components, ancillary reagents required but not provided, and a detailed explanation of the methodology, including molecular target(s) for each analyte, design of target detection reagents, rationale for target selection, limiting factors of the device (
e.g., saturation level of hybridization and maximum amplification and detection cycle number, etc.), internal and external controls, and computational path from collected raw data to reported result (e.g., how collected raw signals are converted into a reported signal and result), as applicable.(v) A detailed description of device software, including software applications and hardware-based devices that incorporate software. The detailed description must include documentation of verification, validation, and hazard analysis and risk assessment activities, including an assessment of the impact of threats and vulnerabilities on device functionality and end users/patients as part of cybersecurity review.
(vi) For devices intended for the detection and identification of microbial agents for which an FDA recommended reference panel is available, design verification and validation must include the performance results of an analytical study testing the FDA recommended reference panel of characterized samples. Detailed documentation must be kept of that study and its results, including the study protocol, study report for the proposed intended use, testing results, and results of all statistical analyses.
(vii) For devices with an intended use that includes detection of Influenza A and Influenza B viruses and/or detection and differentiation between the Influenza A virus subtypes in human clinical specimens, the design verification and validation must include a detailed description of the identity, phylogenetic relationship, or other recognized characterization of the Influenza A and B viruses that the device is designed to detect, a description of how the device results might be used in a diagnostic algorithm and other measures that might be needed for a laboratory identification of Influenza A or B virus and of specific Influenza A virus subtypes, and a description of the clinical and epidemiological parameters that are relevant to a patient case diagnosis of Influenza A or B and of specific Influenza A virus subtypes. An evaluation of the device compared to a currently appropriate and FDA accepted comparator method. Detailed documentation must be kept of that study and its results, including the study protocol, study report for the proposed intended use, testing results, and results of all statistical analyses.
(5) When applicable, performance results of the analytical study testing the FDA recommended reference panel described in paragraph (b)(4)(vi) of this section must be included in the device's labeling under § 809.10(b) of this chapter.
(6) For devices with an intended use that includes detection of Influenza A and Influenza B viruses and/or detection and differentiation between the Influenza A virus subtypes in human clinical specimens in addition to detection of SARS-CoV-2 and similar microbial agents, the required labeling under § 809.10(b) of this chapter must include the following:
(i) Where applicable, a limiting statement that performance characteristics for Influenza A were established when Influenza A/H3 and A/H1-2009 (or other pertinent Influenza A subtypes) were the predominant Influenza A viruses in circulation.
(ii) Where applicable, a warning statement that reads if infection with a novel Influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent influenza viruses and sent to State or local health departments for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.
(iii) Where the device results interpretation involves combining the outputs of several targets to get the final results, such as a device that both detects Influenza A and differentiates all known Influenza A subtypes that are currently circulating, the device's labeling must include a clear interpretation instruction for all valid and invalid output combinations, and recommendations for any required followup actions or retesting in the case of an unusual or unexpected device result.
(iv) A limiting statement that if a specimen yields a positive result for Influenza A, but produces negative test results for all specific influenza A subtypes intended to be differentiated (
i.e., H1-2009 and H3), this result requires notification of appropriate local, State, or Federal public health authorities to determine necessary measures for verification and to further determine whether the specimen represents a novel strain of Influenza A.(7) If one of the actions listed at section 564(b)(1)(A) through (D) of the Federal Food, Drug, and Cosmetic Act occurs with respect to an influenza viral strain, or if the Secretary of Health and Human Services determines, under section 319(a) of the Public Health Service Act, that a disease or disorder presents a public health emergency, or that a public health emergency otherwise exists, with respect to an influenza viral strain:
(i) Within 30 days from the date that FDA notifies manufacturers that characterized viral samples are available for test evaluation, the manufacturer must have testing performed on the device with those influenza viral samples in accordance with a standardized protocol considered and determined by FDA to be acceptable and appropriate.
(ii) Within 60 days from the date that FDA notifies manufacturers that characterized influenza viral samples are available for test evaluation and continuing until 3 years from that date, the results of the influenza emergency analytical reactivity testing, including the detailed information for the virus tested as described in the certificate of authentication, must be included as part of the device's labeling in a tabular format, either by:
(A) Placing the results directly in the device's labeling required under § 809.10(b) of this chapter that accompanies the device in a separate section of the labeling where analytical reactivity testing data can be found, but separate from the annual analytical reactivity testing results; or
(B) In a section of the device's label or in other labeling that accompanies the device, prominently providing a hyperlink to the manufacturer's public website where the analytical reactivity testing data can be found. The manufacturer's website, as well as the primary part of the manufacturer's website that discusses the device, must provide a prominently placed hyperlink to the website containing this information and must allow unrestricted viewing access.