(105 days)
Not Found
No
The description focuses on standard molecular diagnostic techniques (nucleic acid amplification, target capture, TMA, DKA) and does not mention AI or ML. The result determination is based on a cut-off and kinetic curve type, which are not indicative of AI/ML.
No.
The device is an in vitro diagnostic test intended for the qualitative detection of RNA from SARS-CoV-2. It aids in diagnosis and does not provide therapy or treatment.
Yes
The "Intended Use / Indications for Use" section explicitly states that the Aptima SARS-CoV-2 Assay is "a nucleic acid amplification in vitro diagnostic test" and is "intended for use as an aid in the diagnosis of COVID-19."
No
The device is an in vitro diagnostic test that involves physical components for sample collection, processing, and analysis (reagents, magnetic microparticles, probes, luminometer). While software is likely involved in controlling the automated system and analyzing results, the core of the device is a hardware-dependent assay.
Yes, this device is an IVD (In Vitro Diagnostic).
The document explicitly states in the "Intended Use / Indications for Use" section: "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)..."
Furthermore, the "Device Description" section also refers to it as: "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..."
These statements clearly identify the device as an in vitro diagnostic test.
N/A
Intended Use / Indications for Use
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.
Product codes (comma separated list FDA assigned to the subject device)
OOX
Device Description
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.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
nasopharyngeal (NP) swab, anterior nasal (AN) swab
Indicated Patient Age Range
patients with signs and symptoms of COVID-19 (all ages)
Intended User / Care Setting
Professional use
Description of the training set, sample size, data source, and annotation protocol
Not Found
Description of the test set, sample size, data source, and annotation protocol
Clinical Study 1: Prospective Clinical Study - Nasopharyngeal Swab Specimens in UTM/VTM
- Sample Size: 1495 evaluable NP swab specimens (1195 fresh, 300 frozen).
- Data Source: Remnant NP swab specimens from male and female individuals of all ages exhibiting signs and/or symptoms of respiratory infection consistent with COVID-19, collected prospectively at four US pediatric/adolescent, private and/or university hospitals.
- Annotation Protocol: The Aptima SARS-CoV-2 Assay was evaluated for SARS-CoV-2 performance by comparing its results from NP swab specimens in UTM/VTM to a composite comparator algorithm (CCA) consisting of two highly sensitive US FDA EUA SARS-CoV-2 molecular tests and a validated PCR followed by bi-directional sequencing (PCR/BDS) assay. A final CCA result was assigned when two of the three comparator assay results were in concordance. NP specimens that obtained discordant results underwent additional testing with a US FDA EUA SARS-CoV-2 molecular test, volume permitting.
Clinical Study 2: Prospective Clinical Study – Anterior Nasal Swab Specimens in UTM/VTM and eSTM (RespDirect Collection Kit)
- Sample Size: 2177 evaluable individuals (1159 with evaluable anterior nasal swab specimens in UTM/VTM, and 1018 with evaluable nasal swab specimens in eSTM), from a total of 2295 non-withdrawn subjects enrolled.
- Data Source: Male and female individuals of all ages exhibiting signs and/or symptoms of respiratory infection consistent with COVID-19, enrolled at nine geographically and ethnically diverse US sites during the 2022-2023 respiratory season. Two anterior nasal swab specimens were prospectively collected from each individual: one specimen collected by a healthcare professional (HCP) and stored in UTM/VTM; one specimen collected by the patient (under HCP supervision) or the HCP using either a synthetic flocked swab and stored in UTM/VTM or using the RespDirect flocked swab and stored in a Direct Capture Tube containing eSTM.
- Annotation Protocol: The Aptima SARS-CoV-2 Assay was evaluated for SARS-CoV-2 performance by comparing its results from anterior nasal swab specimens in UTM/VTM or in eSTM to a composite comparator algorithm (CCA) consisting of two highly sensitive US FDA EUA SARS-CoV-2 molecular tests and a validated PCR/BDS assay. A final CCA result was assigned when two of the three comparator assay results were in concordance.
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Analytical (Non-Clinical) Studies:
- Analytical Sensitivity - Limit of Detection (LoD):
- Study Type: LoD determination and verification using inactivated cultured SARS-CoV-2 virus and WHO International Standard for SARS-CoV-2.
- Sample Size: Ten replicates for initial LoD determination, additional minimum 20 replicates for verification for cultured virus. Minimum 24 replicates for WHO International Standard, confirmed with an additional 24 replicates.
- Key Results: LoD determined to be 0.01 TCID50/mL in the test sample (0.026 TCID50/mL in the neat, unprocessed sample) for cultured virus. For WHO International Standard, 87.5 IU/mL was the lowest concentration at which ≥95% detection was observed. LoD confirmation with RespDirect Collection Kit also showed ≥95% detection at 87.5 IU/mL.
- Reactivity - Wet Testing:
- Study Type: Evaluation of detection capability for various SARS-CoV-2 virus strains.
- Sample Size: Each strain tested in triplicate.
- Key Results: 100% positivity was observed at concentrations ranging from 0.015 TCID50/mL to 0.151 TCID50/mL for various strains, including multiple variants of concern.
- Reactivity - In silico Analysis:
- Study Type: In silico evaluation of assay target capture oligos, amplification primers, and detection probes against NCBI and GISAID gene databases.
- Key Results: Predicted to detect 99.98% (2,136,815/2,137,175 sequences) of all sequences evaluated, including all lineages and variants of public health interest identified as of January 31, 2024.
- Analytical Specificity and Microbial Interference:
- Study Type: Evaluation of cross-reactivity and microbial interference by closely related and non-targeted organisms.
- Sample Size: Panels of 48 organisms tested in triplicate.
- Key Results: No cross-reactivity or microbial interference observed for any of the 48 organisms tested at indicated concentrations.
- Interfering Substances:
- Study Type: Evaluation of endogenous and exogenous substances.
- Key Results: No impact on performance seen for any substances at tested concentrations.
- Carryover Contamination:
- Study Type: Assessment of carryover rate.
- Sample Size: 588 negative and positive valid tests across three Panther systems (294 of each).
- Key Results: Observed carryover rate of 0% (0/294).
- Assay Precision:
- Study Type: Within-lab precision evaluation.
- Sample Size: 4-member panel (Negative, High Negative (0.1X LoD), Low Positive (1X LoD), Moderate Positive (5X LoD)), tested in three replicates per run for a total of 108 replicates per panel.
- Key Results: Agreement with expected results was 100% in the Negative, Low Positive, and Moderate Positive panel members. High Negative panel (10X below LoD) had 68/108 (63%) positive results. Total SARS-CoV-2 signal variability (as %CV) ranged from 2.75% to 3.84% for Negative, Low Positive, and Moderate Positive panels.
- Collection Device Equivalency:
- Study Type: Equivalence between NP specimens collected into VTM/UTM and NP and AN swab specimens collected in RespDirect (eSTM).
- Sample Size: 150 negative specimens, 50 at 2X LoD, and 50 at 5X LoD per collection device for contrived panels.
- Key Results: Demonstrated similar agreement comparable between the two collection devices.
- Reproducibility:
- Study Type: Multi-site reproducibility evaluation.
- Sample Size: One negative and two positive panel members (Low Positive 1-2X LoD, Moderate Positive 3-5X LoD), tested at three US sites, six operators, over at least five days. Each run had three replicates of each panel member.
- Key Results: Agreement with expected results was 100% for all panel members. Total SARS-CoV-2 signal variability (as %CV) was ≤7.93% for all positive panel members.
Clinical Studies:
- Clinical Study 1: Prospective Clinical Study - Nasopharyngeal Swab Specimens in UTM/VTM
- Study Type: Prospective, multicenter clinical study.
- Sample Size: 1495 evaluable NP swab specimens.
- Key Results:
- Overall Positive Percent Agreement (PPA): 95.4% (90.3-97.9% CI)
- Overall Negative Percent Agreement (NPA): 99.5% (98.9-99.8% CI)
- Clinical Study 2: Prospective Clinical Study – Anterior Nasal Swab Specimens in UTM/VTM and eSTM (RespDirect Collection Kit)
- Study Type: Prospective, multicenter clinical study.
- Sample Size: 1159 evaluable anterior nasal swab specimens in UTM/VTM, and 1018 with evaluable nasal swab specimens in eSTM.
- Key Results:
- UTM/VTM: PPA 96.5% (92.1-98.5% CI), NPA 97.6% (96.5-98.4% CI)
- RespDirect eSTM: PPA 100% (96.6-100% CI), NPA 98.0% (96.9-98.7% CI)
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Clinical Study 1: NP Swab Specimens
- Positive Percent Agreement (PPA): Overall 95.4% (TP/ (TP+FN) = 124/130). 95% CI: 90.3-97.9.
- Negative Percent Agreement (NPA): Overall 99.5% (TN/ (FP+TN) = 1358/1365). 95% CI: 98.9-99.8.
Clinical Study 2: Anterior Nasal Swab Specimens
- UTM/VTM:
- Positive Percent Agreement (PPA): 96.5% (TP/ (TP+FN) = 138/143). 95% CI: 92.1-98.5.
- Negative Percent Agreement (NPA): 97.6% (TN/ (FP+TN) = 992/1016). 95% CI: 96.5-98.4.
- RespDirect eSTM:
- Positive Percent Agreement (PPA): 100% (TP/ (TP+FN) = 108/108). 95% CI: 96.6-100.
- Negative Percent Agreement (NPA): 98.0% (TN/ (FP+TN) = 892/910). 95% CI: 96.9-98.7.
Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.
Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).
Not Found
§ 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.
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Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). On the left 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.
February 13, 2025
Hologic, Inc. Katerina Capkova Regulatory Affairs Manager 10210 Genetic Center Dr. San Diego, California 92121
Re: K243396
Trade/Device Name: Aptima SARS-CoV-2 Assay Regulation Number: 21 CFR 866.3981 Regulation Name: 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 Regulatory Class: Class II Product Code: OOX Dated: October 31, 2024 Received: October 31, 2024
Dear Katerina Capkova:
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.
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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).
Your device is also subject to, among other requirements, the Quality System (OS) 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 (OS) 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.
All medical devices, including Class I and unclassified devices and combination product device constituent parts are required to be in compliance with the final Unique Device Identification System rule ("UDI Rule"). The UDI Rule requires, among other things, that a device bear a unique device identifier (UDI) on its label and package (21 CFR 801.20(a)) unless an exception or alternative applies (21 CFR 801.20(b)) and that the dates on the device label be formatted in accordance with 21 CFR 801.18. The UDI Rule (21 CFR 830.300(a) and 830.320(b)) also requires that certain information be submitted to the Global Unique Device Identification Database (GUDID) (21 CFR Part 830 Subpart E). For additional information on these requirements, please see the UDI System webpage at https://www.fda.gov/medical-device-advicecomprehensive-regulatory-assistance/unique-device-identification-system-udi-system.
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 medical devices and radiation-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-regulatory
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assistance/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,
Anna M. Mielech -S
Anna Mielech, PhD. Deputy Branch Chief (Acting) Viral Respiratory and HPV 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) K243396
Device Name Aptima SARS-CoV-2 Assay
Indications for Use (Describe)
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.
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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510(k) SUMMARY
Aptima SARS-CoV-2 Assay
Hologic, Inc. I. SUBMITTER: 10210 Genetic Center Drive San Diego, CA 92121
- Contact Person: Katerina Capkova, PhD Regulatory Affairs Manager katerina.capkova@hologic.com Phone: 858.410.8167 Fax: N/A
Date Prepared: February 11, 2025
II. DEVICE
| Proprietary Name: | Aptima SARS-CoV-2 Assay |
|---|---|
| Classification Name: | Respiratory Specimen Nucleic Acid SARS-CoV-2 Test |
| Regulation Number: | 21 CFR 866.3981 |
| Regulatory Class: | Class II |
| Product Code: | QQX |
III. PREDICATE DEVICE
The predicate device for the Aptima SARS-CoV-2 Assay is the BioFire COVID-19 Test 2 (K211079; cleared November 1, 2021, BioFire Defense, LLC).
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IV. DEVICE DESCRIPTION
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
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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.
Assay Components
The Aptima SARS-CoV-2 Assay is available in 100-test and 250-test kit configuration. In both cases, the assay kit consists of two boxes containing nine reagents which are required for sample processing. In addition, the controls kit contains the positive and negative controls. RespDirect Collection Kit is an ancillary component of the assay. The Panther Fusion Specimen Lysis Tubes are also an ancillary kit to this assay required for processing of Viral Transport Media/Universal Transport Media (VTM/UTM) specimens prior to testing on the Panther/Panther Fusion system. In addition, there is one ancillary kit (Aptima Assay Fluids Kit) which is universally used with other commercialized Aptima assays.
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Table 1: Reagents Required to Perform the Aptima SARS-CoV-2 Assay
| Box | Components Description |
|---|---|
| Assay Kit- | |
| Refrigerated | Amplification Reagent |
| Enzyme Reagent | |
| Promoter Reagent | |
| Internal Control | |
| Assay Kit-Room | |
| Temperature | Amplification Reconstitution Solution |
| Enzyme Reconstitution Solution | |
| Promoter Reconstitution Solution | |
| Selection Reagent | |
| Target Capture Reagent | |
| Controls Kit- | |
| Refrigerated | Negative Control |
| Positive Control |
Table 2: Ancillary and Collection Kits Required to Perform the Aptima SARS-CoV-2 Assay
| RespDirect Collection Kit |
|---|
| Panther Fusion Specimen Lysis Tubes |
| Aptima Assay Fluids Kit |
Instrumentation
The Aptima SARS-CoV-2 Assay has been designed for and validated on the Panther/Panther Fusion system. The Panther Panther Fusion system is an integrated hardware and software system that together with the Aptima SARS-CoV-2 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
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.
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VI. COMPARISON OF TECHNOLOGICAL CHARACTERISTICS WITH THE PREDICATE DEVICE
A comparison of the Aptima SARS-CoV-2 Assay to the predicate BioFire COVID-19 Test 2 (K211079) is summarized in Table 3.
Table 3: Comparison of Similarities and Differences Between the Subject Device (Aptima SARS-CoV-2 Assay) and the Predicate Device (BioFire COVID-19 Test 2)
| Item | Aptima
SARS-CoV-2 Assay
(Subject Device) | BioFire COVID-19 Test 2
(Predicate Device)
K211079 |
|-----------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Prescription/over-
the-counter use | Prescription Only | Same |
| Patient
Population | Individuals with signs and symptoms of COVID-19 | Same |
| Specimen Types | Nasopharyngeal (NP) and nasal swab specimens | Nasopharyngeal (NP) swab specimens |
| Intended User | Professional use | Same |
| Technology
Principle of
Operation | Transcription-mediated amplification NAAT | Reverse transcriptase multiplexed polymerase chain
reaction test |
| Organisms
Detected | SARS-CoV-2 | Same |
| Assay Controls | Internal and run controls | Internal and external controls |
| Platform | Automated nucleic acid amplification platform.
Uses Panther/Panther Fusion system for all steps
including nucleic acid extraction, amplification,
detection, and result processing. | Automated nucleic acid amplification platform.
Uses BioFire FilmArray 2.0 or BioFire FilmArray
Torch systems including integrated sample preparation,
amplification, detection, and analysis. |
| Intended Use | 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 BioFire COVID-19 Test 2 is a qualitative nested
multiplexed RT-PCR in vitro diagnostic test intended
for use with the BioFire FilmArray 2.0 and BioFire
FilmArray Torch Systems. The BioFire COVID-19 Test
2 detects nucleic acids from severe acute respiratory
syndrome coronavirus 2 (SARS-CoV-2) in
nasopharyngeal swabs (NPS) from symptomatic
individuals suspected of COVID-19 by their healthcare
provider.
Results are for the identification of SARS-CoV-2 RNA.
The SARS-CoV-2 RNA is generally detectable in NPS
specimens during the acute phase of infection. Positive
results are indicative of the presence of SARS-CoV-2
RNA; clinical correlation with patient history and other
diagnostic information is necessary to determine patient
infection status. Positive results do not rule out co-
infection with other pathogens.
Results are meant to be used in conjunction with other
clinical, epidemiologic, and laboratory data, in
accordance with the guidelines provided by the relevant
public health authorities. The BioFire COVID-19 Test 2
is intended for use by trained medical and laboratory
professionals in a laboratory setting or under the
supervision of a trained laboratory professional. |
| Time to Obtain
Test Results | Approximately 2.5 hours | Approximately 45 minutes |
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VII. PERFORMANCE DATA
The following performance data (analytical and clinical) were provided in support of the substantial equivalence determination.
Brief Description of Analytical (Non-Clinical) Studies
The following analytical studies (non-clinical) were conducted to support the clearance of the Aptima SARS-CoV-2 Assay on the Panther/Panther Fusion System.
Analytical Sensitivity - Limit of Detection (LoD)
The analytical sensitivity (limit of detection or LoD) of the Aptima SARS-CoV-2 Assay was determined by testing dilutions of processed negative clinical NP swab VTM/UTM matrix spiked with inactivated cultured SARS-CoV-2 virus (USA-WA1/2020; BEI Resources; NR-52281) and WHO International Standard for SARS-CoV-2, NIBSC (20/146). For the cultured virus, ten replicates of each serial dilution were evaluated for each of two assay reagent lots across two Panther systems. The LoD was determined to be 0.01 TCID55/mL in the test sample (0.026 TCID50/mL in the neat, unprocessed sample) and verified by testing an additional minimum 20 replicates with one assay reagent lot. For the WHO International Standard, a minimum of 24 replicates were tested with each of the three reagent lots using Probit analysis for each lot and was confirmed with an additional 24 replicates using a single lot. The lowest concentration at which ≥95% detection was observed was 87.5 IU/mL . LoD confirmation was also performed with the RespDirect Collection Kit at 24 replicates with a single reagent lot and ≥95% detection was determined to be 87.5 IU/mL.
Note: The stated LoDs pertain to the concentrations in the tubes loaded onto the instrument. For samples collected in VTM/UTM, this is the concentration in the processed sample in an SLT. For samples collected using the RespDirect Collection kit, this is the concentration in the Enhanced Direct Load tube (RespDirect Collection Kit).
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Reactivity - Wet Testing
The reactivity of the Aptima SARS-CoV-2 Assay was determined by testing virus strains in processed negative clinical NP swab VTM/UTM matrix. Each strain was tested in triplicate at 3X LoD with one reagent lot. For strains not detected at 3X LoD, additional testing at higher concentrations was performed until 100% positivity was observed. Table 4 shows the lowest concentration of each strain in which 100% positivity was observed.
| Description | Concentration |
|---|---|
| USA-WA1/2020* | 0.03 TCID50/mL |
| USA-CA1/2020 | 0.03 TCID50/mL |
| USA-AZ1/2020 | 0.101 TCID50/mL |
| USA-WI1/2020 | 0.03 TCID50/mL |
| USA/OR-OHSU-PHL00037/2021 B.1.1.7 | 0.03 TCID50/mL |
| Uganda/MUWRP-20200195568/2020 A.23.1 | 0.03 TCID50/mL |
| USA/PHC658/2021 B.1.617.2 | 0.03 TCID50/mL |
| USA/MD-HP05285/2021 B.1.617.2 | 0.03 TCID50/mL |
| USA/CA/VRLC009/2021 B.1.427 | 0.03 TCID50/mL |
| USA/CA/VRLC012/2021 P.2 | 0.03 TCID50/mL |
| USA/MD-HP03056/2021 B.1.525 | 0.03 TCID50/mL |
| USA/CA-Stanford-15_S02/2021 B.1.617.1 | 0.03 TCID50/mL |
| Peru/un-CDC-2-4069945/2021 C.37 | 0.03 TCID50/mL |
| USA/MD-HP20874/2021 B.1.1.529 | 0.03 TCID50/mL |
| USA/GA-EHC-2811C/2021 B.1.1.529 | 0.03 TCID50/mL |
| USA/MD-HP30386/2022 BA.4 | 0.03 TCID50/mL |
| USA/COR-22-063113/2022 BA.5 | 0.03 TCID50/mL |
| South Africa/CERI-KRISP-K040013/2022 BA.5 | 0.03 TCID50/mL |
| USA/MD-HP38861/2022 BQ.1.1 | 0.03 TCID50/mL |
| USA/MD-HP40900/2022 XBB.1.5 | 0.101 TCID50/mL |
| USA/MD-HP47865/2023 XXB.2.3 | 0.03 TCID50/mL |
| USA/MD-HP46933/2023 EG.1.2 | 0.03 TCID50/mL |
| USA/MD-HP47946/2023 EG.5.1 | 0.03 TCID50/mL |
Table 4: Analytical Reactivity Summary for SARS-CoV-2
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| USA/CA-Stanford-139_S35/2023 XBB.1.9 | 0.101 TCID50/mL |
|---|---|
| USA/CA-Stanford-139_S23/2023 XBB.1.16 | 0.101 TCID50/mL |
| USA/MI-UM-10052670540/2023 BA.2.86 | 0.102 TCID50/mL |
| USA/New York-PV96109/2023 JN.1 | 0.151 TCID50/mL |
| USA/MD-HP49152/2023 HV.1 | 0.015 TCID50/mL |
*Strain used to establish LoD.
1In silico analysis showed 100% homology to amplification regions.
21n silico analysis identified a single mismatch in the probe oligo for one region. Due to the mismatch and 100% homology to the second region, detection is not expected to be impacted. Virus stock degradation or error in TCID50mL quantification may have impacted the concentration at 100% detection.
Reactivity-In silico Analysis
The inclusivity of the Aptima SARS-CoV-2 Assay was evaluated using in silico analysis of the assay target capture oligos, amplification primers, and detection probes for the SARS-CoV-2 target systems in relation to sequences available in the NCBI and GISAID gene databases. Any sequence with missing or ambiguous sequence information was removed from the analysis for that region. Based on the in silico analysis of GISAID and NCBI sequences available for SARS-CoV-2 (10% random sampling of 16.553.661 million sequences up to July, 31, 2023 and all 508,436 sequences August 1, 2023 - January 31, 2024), the Aptima SARS-CoV-2 Assay is predicted to detect 99.98% (2,136,815/2,137,175 sequences) of all sequences evaluated.
The sequences evaluated included lineages and variants of concern (VOC) or variants under investigation (VUI) that may have important epidemiological, immunological, or pathogenic properties from the public health perspective. All lineages and variants of public health interest identified as of January 31, 2024 are predicted to be detected; new sequences and variants will continue to be monitored for impacts on detection by the Aptima SARS-CoV-2 Assay.
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Analytical Specificity and Microbial Interference
Analytical specificity (cross-reactivity) and microbial interference with the Aptima SARS-CoV-2 Assay were evaluated in the presence of closely related and non-targeted organisms. Panels consisting of 48 organisms (6) were tested in processed negative clinical NP swab VTM/ UTM matrix in the absence or presence of 3X LoD SARS-CoV-2. Bacteria were tested at 106 CFU/mL and viruses were tested at 105 TCID50/mL, except where noted. No crossreactivity or microbial interference was observed for any of the 48 organisms tested on the Aptima SARS-CoV-2 Assay at the indicated concentrations.
| Microorganism | Concentration¹ | Microorganism | Concentration¹ |
|---|---|---|---|
| Adenovirus 1 | 1x105 TCID50/mL | Aspergillus fumigatus | 1x106 CFU/mL |
| Adenovirus 7a | 1x105 TCID50/mL | Bordetella parapertussis | 1x106 CFU/mL |
| CMV Strain AD 169 | 5x103 TCID50/mL | Bordetella pertussis | 1x106 CFU/mL |
| EBV | 1x105 TCID50/mL | Candida albicans | 1x106 CFU/mL |
| Enterovirus Type 71 | 1x105 TCID50/mL | Chlamydia pneumoniae | 1x106 CFU/mL |
| Human coronavirus 229E | 1x105 TCID50/mL | Corynebacterium diphtheriae | 1x106 CFU/mL |
| Human coronavirus OC43 | 1x105 TCID50/mL | Escherichia coli | 1x106 CFU/mL |
| Human coronavirus HKU12 | 1x106 copies/mL | Fusobacterium necrophorum | 1x106 CFU/mL |
| Human coronavirus NL63 | 1x104 TCID50/mL | Haemophilus influenzae | 1x106 CFU/mL |
| Human Metapneumovirus | |||
| (hMPV) | 1x106 TCID50/mL | Lactobacillus plantarum | 1x106 CFU/mL |
| Influenza A | 1x105 TCID50/mL | Legionella pneumophila | 1x106 CFU/mL |
| Influenza B | 2x103 TCID50/mL | Moraxella catarrhalis | 1x106 CFU/mL |
| Measles | 1x105 TCID50/mL | Mycobacterium tuberculosis | 1x106 CFU/mL |
| MERS-coronavirus | 1x104 TCID50/mL | Mycoplasma genitalium | 1x106 CFU/mL |
| Mumps | 1x105 TCID50/mL | Mycoplasma pneumoniae | 1x106 CFU/mL |
| Parainfluenza virus 1 | 1x105 TCID50/mL | Neisseria gonorrhoeae | 1x106 CFU/mL |
| Parainfluenza virus 2 | 1x105 TCID50/mL | Neisseria meningitides | 1x106 CFU/mL |
| Parainfluenza virus 3 | 1x105 TCID50/mL | Neisseria mucosa | 1x106 CFU/mL |
| Parainfluenza virus 4 | 1x103 TCID50/mL | Pneumocystis jirovecii (PJP) | 1x106 nuclei/mL |
Table 5: Aptima SARS-CoV-2 Analytical Specificity and Microbial Interference Microorganisms
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| Respiratory syncytial virus | 1x105 TCID50/mL | Pseudomonas aeruginosa | 1x106 CFU/mL |
|---|---|---|---|
| Rhinovirus | 1x104 TCID50/mL | Staphylococcus aureus | 1x106 CFU/mL |
| SARS-coronavirus2 | 1x106 copies/mL | Staphylococcus epidermis | 1x106 CFU/mL |
| Varicella Zoster Virus | 1x104 TCID50/mL | Streptococcus pneumoniae | 1x106 CFU/mL |
| Pooled human nasal wash3 – to represent diverse microbial flora in human respiratory tract | N/A | Streptococcus pyogenes | 1x106 CFU/mL |
| Streptococcus salivaris | 1x106 CFU/mL |
1CFU = Colony Forming Units; TCID50 = Median Tissue Culture Infectious Dose
²Cultured virus and whole genome purified nucleic acid for Human coronavirus were not readily available at the time testing was performed. Human coronavins IVTs corresponding to the ORFIab gene regions targeted by the assay were used to evaluate cross-reactivity and microbial interference.
31n place of evaluating pooled human nasal wash, testing of 30 individual negative clinical NP swab speciforned to represent diverse microbial flora in the human respiratory tract.
Interfering Substances
Interfering endogenous and exogenous substances (mucin, whole blood, potential medications and over-the-counter products) that may be present in the samples were evaluated in the Aptima SARS-CoV-2 assay. Clinically relevant concentrations of potentially interfering substances were added to pooled clinical negative NP swab VTM/UTM matrix and tested in the absence and presence of SARS-CoV-2 inactivated virus at 3X LoD. The substances and concentrations are shown in Table 6. No impact on the performance of the Aptima SARS-CoV-2 Assay was seen for any of the substances at the concentration tested.
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| Table 6: Potentially Interfering Substances | ||
|---|---|---|
| Substance Type | Substance Name | Active Ingredient(s) | Highest Test
Concentration |
|--------------------------|---------------------------|-------------------------------------------------------------------------------|-------------------------------|
| Endogenous | Mucin | Purified mucin protein | 60 µg/mL |
| Endogenous | Blood (human) | N/A | 2% v/v |
| Nasal sprays or
drops | Neo-Synephrine® | Phenylephrine | 15% v/v |
| Nasal sprays or
drops | Anefrin | Oxymetazoline | 15% v/v |
| Nasal sprays or
drops | Saline | Sodium chloride | 15% v/v |
| Nasal corticosteroids | Ventolin HFA2 | Albuterol | 45 ng/mL |
| Nasal corticosteroids | QVAR® Beconase AQ2 | Beclomethasone | 15 ng/mL |
| Nasal corticosteroids | Dexacort2 | Dexamethasone | 12 µg/mL |
| Nasal corticosteroids | Flonase | Fluticasone | 5% v/v |
| Nasal corticosteroids | Nasacort | Triamcinolone | 5% v/v |
| Nasal corticosteroids | Rhinocort | Budesonide | 5% v/v |
| Nasal corticosteroids | Nasonex2 | Mometasone | 0.5 ng/mL |
| Nasal corticosteroids | AEROSPAN®2 | Flunisolide | 9.9 µg/mL |
| Nasal gel | Zicam® (Allergy Relief) | Luffa opperculata, Galphimia,
Glauca, Histaminum
hydrochloricum, Sulfur | 5% v/v |
| Throat lozenges | Cepacol Extra Strength | Benzocaine, Menthol | 0.7 mg/mL |
| Throat lozenges | Cold-Eeze throat lozenge | Zinc gluconate | 0.7 mg/mL |
| Anti-viral drugs | Relenza2 | Zanamivir | 3.3 mg/mL |
| Anti-viral drugs | TamiFlu2 | Oseltamivir | 399 ng/mL |
| Anti-viral drugs | Virazole2 | Ribavirin | 10.5 µg/mL |
| Antibiotic, nasal | Bactroban cream2 | Mupirocin | 1.6 µg/mL |
| Antibacterial, | Tobramycin2 | Tobramycin | 33.1 µg/mL |
| Solvent Control | Water | N/A | 5% v/v |
| Solvent Control | Dimethyl Sulfoxide (DMSO) | N/A | 5% v/v |
¹v/v: volume by volume
2Active ingredient tested, not substance
3T wo out of three positive results were observed for the positive pool containing Nasacort and retested and all replicates (3/3) were positive.
Carryover Contamination
The carryover contamination rate of the Aptima SARS-CoV-2 Assay for samples tested was assessed by testing high titer panels consisting of SARS-CoV-2 virus in negative clinical NP swab VTM/UTM matrix spiked 100 TCID56/mL (10,000 times the assay LoD). Positive panels were tested in a checkerboard pattern, alternating with negative panels. Testing consisted of 588
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negative and positive valid tests across three Panther systems. The Aptima SARS-CoV-2 Assay observed a carryover rate of 0% (0/294).
Assay Precision
The Aptima SARS-CoV-2 Assay within-lab precision was evaluated with a 4-member panel consisting of virus in negative clinical NP swab VTM/UTM matrix. The 4-member panel included a Negative, a High Negative (0.1X LoD), a Low Positive (1X LoD) and a Moderate Positive (5X LoD) panels were tested by two operators, using three reagent lots on three Panther systems over six days, at one site. Two runs were performed per operator per day for a total minimum of 36 runs. Each of the four panels was tested in three replicates per run for a total of 108 replicates per panel. The agreement with expected results was 100% in the Negative, Low Positive and Moderate Positive panel members. The High Negative panel member was 10X below the assay LoD, therefore a mix of positive and negative results were expected. This panel had 68/108 (63%) positive results. Agreement with expected results for all four panels is shown in Table 7.
| Panel
Description | Panel
Composition | Panel Conc.
TCID50/mL | Expected
Result | N
Positive | N
Tested | Mean
kRLU | Agreement
w/Expected
(95% CI) |
|----------------------|----------------------|--------------------------|--------------------|---------------|-------------|--------------|-------------------------------------|
| Negative | N/A | N/A | Negative | 0 | 108 | 289 | 100%
(96.6-100) |
| High
Negative | 0.1xLoD | 0.001 | N/A | 68 | 108 | 627 | N/A |
| Low
Positive | 1.0xLoD | 0.01 | Positive | 108 | 108 | 1131 | 100%
(96.6-100) |
| Moderate
Positive | 5.0xLoD | 0.05 | Positive | 108 | 108 | 1147 | 100%
(96.6-100) |
Table 7: Agreement of Aptima SARS-CoV-2 Assay Results with Expected Results
The total SARS-CoV-2 signal variability measured as %CV ranged from 2.75% to 3.84% in Negative, Low Positive, and Moderate Positive panel members. For the sources of variation all six factors evaluated had %CV values