(258 days)
Alinity m EBV is an in vitro polymerase chain reaction (PCR) assay for the quantitation of Epstein-Barr Virus (EBV) DNA in human EDTA plasma on the automated Alinity m System.
Alinity m EBV is intended for use as an aid in the management of EBV in transplant patients. In patients undergoing monitoring of EBV, serial DNA measurements can be used to indicate the need for potential treatment changes and to assess viral response to treatment.
The results from Alinity m EBV must be interpreted within the context of all relevant clinical and laboratory findings.
Alinity m EBV is not cleared for use as a screening test for donors of blood, blood products, or human cells, tissues, and cellular and tissue-based products (HCT/Ps) for EBV.
Alinity m EBV is an in vitro polymerase chain reaction (PCR) assay for the quantitation of EBV DNA in human plasma.
This device is similar to the predicate device originally cleared (K212778) with the exception that the subject device may use MomentaTaq DNA Polymerase as an alternative to KAPA2G DNA Polymerase in the reagent formulation of the assay. This formulation difference does not introduce any changes to sample processing, assay procedure, or data reduction.
Additional studies were initiated to support the formulation of the assay with MomentaTaq DNA Polymerase. Supplemental data from these studies were used with data previously obtained from the analytical and clinical testing studies submitted in K212778.
The steps of the Alinity m EBV consist of sample preparation, real-time PCR assembly, amplification/detection, result calculation, and reporting. All stages of the Alinity m EBV procedure are executed automatically by the Alinity m System. No intermediate processing or transfer steps are performed by the user. The Alinity m System is designed to be a random-access analyzer that can perform the Alinity m EBV assay in parallel with other Alinity m assays on the same instrument.
Alinity m EBV requires three separate assay specific kits as follows:
-
Alinity m EBV AMP Kit (List No. 09N43-095), consisting of 2 types of multi-well assay trays. The amplification trays (AMP TRAY 1) contain lyophilized, unit-dose PCR amplification/detection reagents and lyophilized, unit-dose IC in separate wells, and the activation trays (ACT TRAY 2) contain liquid unit-dose activation reagent. The intended storage condition for the Alinity m EBV AMP Kit is 2°C to 8°C.
-
Alinity m EBV CTRL Kit (List No. 09N43-085), consisting of negative controls, low positive controls, and high positive controls, each supplied as liquid in single-use tubes. The intended storage condition for the Alinity m EBV CTRL Kit is –25°C to –15°C.
-
Alinity m EBV CAL Kit (List No. 09N43-075), consisting of 2 calibrator levels, each supplied as liquid in single-use tubes. The intended storage condition for the Alinity m EBV CAL Kit is –25°C to –15°C.
EBV DNA from human plasma is extracted automatically on-board in the Alinity m System using the Alinity m Sample Prep Kit 2, Alinity m Lysis Solution, and Alinity m Diluent Solution. The Alinity m System employs magnetic microparticle technology to facilitate nucleic acid capture, wash, and elution. The resulting purified nucleic acids are then combined with liquid unit-dose Alinity m EBV activation reagent and lyophilized unit-dose Alinity m EBV amplification/detection reagents and transferred into a reaction vessel. Alinity m Vapor Barrier Solution is then added to the reaction vessel which is then transferred to an amplification/detection unit for PCR amplification, and real-time fluorescence detection of EBV targets.
At the beginning of the Alinity m EBV sample preparation process, a lyophilized unit-dose IC on the AMP Tray is rehydrated by the Alinity m System and delivered into each sample preparation reaction. The IC is then processed through the entire sample preparation and real-time PCR procedure along with the specimens, calibrators, and controls to demonstrate proper sample processing and validity.
The Alinity m EBV amplification/detection reagents consist of enzymes, primers, probes, and activation reagents that enable polymerization and detection.
An EBV calibration curve is required for determination of EBV DNA concentration. Two levels of calibrators are processed through sample preparation and PCR to generate the calibration curve. The concentration of EBV DNA in specimens and controls is then calculated from the stored calibration curve.
Assay controls are tested at or above an established minimum frequency to help ensure that instrument and reagent performance remains satisfactory. During each control event, a negative control, a low-positive control, and a high-positive control are processed through sample preparation and PCR procedures that are identical to those used for specimens.
The Alinity m EBV assay also utilizes the following:
- Alinity m EBV Application Specification File, (List No. 09N43-05B)
- Alinity m System and System Software (List No. 08N53-002)
- Alinity m Sample Prep Kit 2 (List No. 09N12-001)
- Alinity m Specimen Dilution Kit I (List No. 09N50-001)
- Alinity m System Solutions, (List No. 09N20):
- Alinity m Lysis Solution (List No. 09N20-001)
- Alinity m Diluent Solution (List No. 09N20-003)
- Alinity m Vapor Barrier Solution, (List No. 09N20-004)
- Alinity m Tubes and Caps (List No. 09N49):
- Alinity m LRV Tube (List No. 09N49-001)
- Alinity m Transport Tubes Pierceable Capped (List No. 09N49-010)
- Alinity m Transport Tube (List No. 09N49-011)
- Alinity m Pierceable Cap (List No. 09N49-012)
- Alinity m Aliquot Tube (List No. 09N49-013)
This document, K243489, is a 510(k) clearance letter for the Alinity m EBV assay, specifically focusing on the use of MomentaTaq DNA Polymerase as an alternative to KAPA2G DNA Polymerase. The primary goal of the studies described is to demonstrate that the device formulated with MomentaTaq DNA Polymerase performs equivalently to the previously cleared device formulated with KAPA2G DNA Polymerase (K212778).
Here's an analysis of the acceptance criteria and study information provided:
1. Table of Acceptance Criteria and Reported Device Performance
| Performance Characteristic | Acceptance Criteria with MomentaTaq Formulation (Implicitly compared to KAPA2G performance) | Reported Device Performance (MomentaTaq Formulation) |
|---|---|---|
| Limit of Detection (LoD) | Overall detection rate of ≥ 95% at 20 IU/mL (based on previous clearance of K212778). | Overall detection rate of 97.2% at 20 IU/mL. |
| Linear Range | Linear across 50 IU/mL (1.70 Log IU/mL) to 200,000,000 IU/mL (8.30 Log IU/mL). | Linear across 15 IU/mL to 250,000,000 IU/mL (1.18 Log IU/mL to 8.40 Log IU/mL). |
| Precision (Within-laboratory SD) | ≤ 0.25 Log IU/mL for 500 IU/mL to 200,000,000 IU/mL (2.70 Log IU/mL to 8.30 Log IU/mL). | Achieved for all panels in this range (0.06-0.19 Log IU/mL). |
| Precision (Within-laboratory SD) | ≤ 0.50 Log IU/mL for 20 IU/mL to < 500 IU/mL (1.30 Log IU/mL to < 2.70 Log IU/mL). | Achieved for all panels in this range (0.20-0.27 Log IU/mL). |
| Equivalence of Total SD (Precision) | 95% CI for ratio of SD (New/Original) contains 1.00 OR upper bound < 1.00. | All panels were "Yes" (Clinically Acceptable). |
| Equivalence of Total %CV (Precision) | 95% CI for ratio of %CV (New/Original) contains 1.00 OR upper bound < 1.00. | All panels were "Yes" (Clinically Acceptable). |
| Lower Limit of Quantitation (LLoQ) | Reliably quantitated at 50 IU/mL (1.70 Log IU/mL) with TAE and TE ≤ 1.00 Log IU/mL. | Supported at 50 IU/mL (1.70 Log IU/mL) with TAE = 0.59 and TE = 0.57. |
| Reproducibility (Equivalence of Total SD) | 95% CI for ratio of SD (New/Original) contains 1.00 OR ratio < 1.00. | All panels were "Yes" (Clinically Acceptable). |
| Reproducibility (Equivalence of Total %CV) | 95% CI for ratio of %CV (New/Original) contains 1.00 OR ratio < 1.00. | All panels were "Yes" (Clinically Acceptable). |
| Negative Agreement Rate (Reproducibility) | High negative agreement rate for negative samples. | 99.2% (95% CI: 95.4%, 99.9%). |
| Clinical Performance (Method Comparison) | Demonstrates equivalence to the on-market Alinity m EBV assay formulated with KAPA2G. | Deming regression: Slope 1.00, Intercept 0.01, r = 0.993. Mean bias: -0.01 Log IU/mL (95% CI: -0.03, -0.01). |
2. Sample Size Used for the Test Set and Data Provenance
The document describes several additional studies conducted specifically for the MomentaTaq formulation, referring back to the K212778 submission for other characteristics.
- Limit of Detection (LoD):
- Sample Size: For EBV type 1, tested dilutions with 3 lots of amplification reagents across multiple days. For 20 IU/mL, there were 142 replicates (47-48 per lot). Total replicates across all concentrations and lots were higher (e.g., 143 for 15 IU/mL, 142 for 100 IU/mL).
- Data Provenance: Prepared in EBV negative human plasma using the 1st World Health Organization (WHO) International Standard for Epstein-Barr Virus. This suggests controlled laboratory samples.
- Linearity:
- Sample Size: 16 panel members spanning the intended quantitation range.
- Data Provenance: Prepared from plasmid DNA, cultured virus, and clinical specimens. The specific origin country for clinical specimens is not provided, but the use of the WHO International Standard points to internationally recognized reference material.
- Precision:
- Sample Size: 8 panel members, with 270 replicates per panel member (3 lots x 3 operators x 15 days x 2 runs/day x 3 replicates/run, then averaged).
- Data Provenance: Panel members prepared by diluting EBV-positive clinical specimens, EBV cultured virus, or synthetic DNA in human plasma. Quantitation traceable to the 1st WHO International Standard. Specific origin country not provided.
- Lower Limit of Quantitation (LLoQ):
- Sample Size: Calculated from detected samples in the LoD study.
- Data Provenance: Prepared in EBV-negative plasma using the 1st WHO International Standard.
- Clinical Reproducibility:
- Sample Size: 9-member reproducibility panel (8 positive, 1 negative). 1 lot of kits used. Testing performed at 3 clinical sites on 5 non-consecutive days with 2 runs per day. 4 replicates of each panel member tested, ensuring a minimum of 3 valid replicates. For the positive panels, N ranged from 113 to 120. For the negative panel, N = 119 for all sites combined.
- Data Provenance: Positive panel members were prepared using EBV positive clinical specimen, cultured virus, or plasmid DNA diluted in human EDTA plasma. Specific origin country for clinical specimens not provided.
- Clinical Performance (Method Comparison):
- Sample Size: 124 samples with results within the common quantitation range of both assays.
- Data Provenance: Samples used for method comparison between the MomentaTaq formulation and the KAPA2G formulation. Implies ex vivo clinical samples. Specific origin country not provided.
The studies for the MomentaTaq formulation are analytical performance studies and a method comparison to an already cleared device, not direct clinical outcome studies. They appear to be retrospective as they involve testing banked or prepared samples. The reference to "3 clinical sites" for reproducibility suggests a multi-site testing environment, but it's an analytical reproducibility study, not a clinical trial on patient outcomes.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and the Qualifications of Those Experts
The provided document does not mention the number or qualifications of experts used to establish ground truth for the test sets. The ground truth for quantitation appears to be established by:
- Using the 1st World Health Organization (WHO) International Standard for Epstein-Barr Virus for Nucleic Acid Amplification Techniques (NIBSC code 09/260) for LoD, Linearity, and LLoQ.
- Using EBV-positive clinical specimens, cultured virus, or plasmid DNA for precision and reproducibility, with quantitation traceable to the WHO standard.
- The "comparator" assay (on-market Alinity m EBV with KAPA2G DNA Polymerase) as a reference for clinical performance (method comparison).
For a quantitative viral load test, the ground truth is typically defined by reference materials (like WHO standards) or highly characterized samples, rather than by expert consensus on interpretation of results.
4. Adjudication Method for the Test Set
Given that the ground truth for these quantitative assays is based on reference standards and characterized samples, no expert adjudication method (like 2+1, 3+1, none) is described or would typically be applicable in this context. The output is a quantitative value, not a subjective interpretation.
5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was Done
No, an MRMC comparative effectiveness study was not done. This type of study is more common for imaging diagnostics where human readers interpret results with and without AI assistance. This document describes an in vitro diagnostic (IVD) quantitative PCR assay, which is an automated device producing numerical results, not something that human readers interpret in the conventional sense of an MRMC study.
6. If a Standalone Study (Algorithm Only Without Human-in-the-Loop Performance) was Done
Yes, effectively, all the studies described are standalone performance studies. The Alinity m EBV device is an automated system that performs sample preparation, PCR, detection, and result calculation without human intervention in the actual assay run. The performance metrics (LoD, linearity, precision, reproducibility, method comparison) directly evaluate the algorithm's ability to accurately quantify EBV DNA in samples. There is no "human-in-the-loop" component in the performance of the assay itself.
7. The Type of Ground Truth Used
The ground truth used in these studies is primarily based on:
- Reference Materials: The 1st World Health Organization (WHO) International Standard for Epstein-Barr Virus (NIBSC code 09/260) is extensively used for establishing and verifying the quantitative values of the panel members.
- Characterized Samples: EBV positive clinical specimens, cultured virus, and synthetic plasmid DNA where their concentrations are known or precisely determined relative to the WHO standard.
- Comparison to Predicate Device: For the method comparison study, the performance of the new MomentaTaq formulation is benchmarked against the FDA-cleared Alinity m EBV assay (K212778), which serves as a clinical reference.
This is a form of "expert consensus" on the value of reference materials and highly characterized samples, rather than pathology or outcomes data in the usual sense for IVDs.
8. The Sample Size for the Training Set
The document does not specify the sample size used for the training set for the Alinity m EBV assay. Regulatory submissions like 510(k) clearances typically focus on the validation (test set) rather than the development and training data of the algorithm itself, especially for established PCR technologies that do not heavily rely on machine learning requiring distinct training sets in the same manner. This assay is based on PCR technology, where the "training" would involve optimizing reagent concentrations, primer/probe design, and thermocycling conditions, rather than training a machine learning model on a "training set" of patient data.
9. How the Ground Truth for the Training Set Was Established
As mentioned above, specific "training set" details are not provided. For PCR-based assays, the "ground truth" during development (analogous to training) would involve:
- Known Viral Copies: Using synthetic nucleic acids or viral stock solutions with precisely quantified viral load to optimize assay components and parameters.
- Spiked Samples: Spiking known amounts of virus or viral nucleic acid into negative matrix (e.g., human plasma) to simulate different concentrations and evaluate early performance.
- Clinical Samples with Confirmed Status: Using retrospectively collected clinical samples with well-established EBV positive/negative status and viral loads determined by highly accurate laboratory methods or other validated assays to refine and confirm assay performance.
The development process would aim to ensure the assay accurately detected and quantified EBV DNA, aligning with established reference standards and biological knowledge of the virus.
FDA 510(k) Clearance Letter - Alinity m EBV
Page 1
U.S. Food & Drug Administration
10903 New Hampshire Avenue Doc ID# 04017.08.00
Silver Spring, MD 20993
www.fda.gov
July 28, 2025
Abbott Molecular Inc.
Jennifer Peterson
Associate Director Regulatory Affairs
1300 E Touhy Ave
Des Plaines, Illinois 60018
Re: K243489
Trade/Device Name: Alinity m EBV
Regulation Number: 21 CFR 866.3183
Regulation Name: Quantitative Viral Nucleic Acid Test For Transplant Patient Management
Regulatory Class: Class II
Product Code: QLX
Dated: November 8, 2024
Received: November 12, 2024
Dear Jennifer Peterson:
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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K243489 - Jennifer Peterson Page 2
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 (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 (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-safety-reporting-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.
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-devices/device-advice-comprehensive-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-devices/medical-device-safety/medical-device-reporting-mdr-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/medical-devices/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-devices/device-advice-comprehensive-regulatory-
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K243489 - Jennifer Peterson Page 3
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,
Uwe Scherf -S
Uwe Scherf, M.Sc., Ph.D.
Director
Division of Microbiology Devices
OHT7: Office of In Vitro Diagnostics
Office of Product Evaluation and Quality
Center for Devices and Radiological Health
Enclosure
Page 4
FORM FDA 3881 (8/23) Page 1 of 1 PSC Publishing Services (301) 443-6740 EF
DEPARTMENT OF HEALTH AND HUMAN SERVICES
Food and Drug Administration
Indications for Use
Form Approved: OMB No. 0910-0120
Expiration Date: 07/31/2026
See PRA Statement below.
510(k) Number (if known): K243489
Device Name: Alinity m EBV
Indications for Use (Describe)
Alinity m EBV is an in vitro polymerase chain reaction (PCR) assay for the quantitation of Epstein-Barr Virus (EBV) DNA in human EDTA plasma on the automated Alinity m System.
Alinity m EBV is intended for use as an aid in the management of EBV in transplant patients. In patients undergoing monitoring of EBV, serial DNA measurements can be used to indicate the need for potential treatment changes and to assess viral response to treatment.
The results from Alinity m EBV must be interpreted within the context of all relevant clinical and laboratory findings.
Alinity m EBV is not cleared for use as a screening test for donors of blood, blood products, or human cells, tissues, and cellular and tissue-based products (HCT/Ps) for EBV.
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)
CONTINUE ON A SEPARATE PAGE IF NEEDED.
This section applies only to requirements of the Paperwork Reduction Act of 1995.
DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.
The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:
Department of Health and Human Services
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Office of Chief Information Officer
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"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."
Page 5
510(k) Summary
Table of Contents
1.0 510(k) Summary ...................................................................................................... 2
- 1.1 Submitter.......................................................................................................... 2
- 1.2 Device Information.......................................................................................... 2
- 1.3 Predicate Device .............................................................................................. 2
- 1.4 Device Description .......................................................................................... 2
- 1.5 Intended Use .................................................................................................... 6
- 1.6 Similarities and Differences to Predicate Device ............................................ 6
- 1.7 Performance Data ............................................................................................ 9
- 1.8 Conclusions Drawn from the Studies............................................................ 22
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1.0 510(k) Summary
This 510(k) summary of safety and effectiveness information is being submitted in accordance with the requirement of 21 CFR Section 807.92(c).
1.1 Submitter
Applicant Name and Address: Abbott Molecular Inc.
1300 E. Touhy Avenue
Des Plaines, IL 60018
Contact Person: John Bates
Director Regulatory Affairs
Abbott Molecular, Inc.
1300 E. Touhy Avenue
Des Plaines, IL 60018
Phone: 224-361-7007
Fax: 224-361-7269
Date Prepared: October 15, 2024
1.2 Device Information
| Trade Name | Regulation Name | Product Code | Regulation No. | Class |
|---|---|---|---|---|
| Alinity m EBV | Quantitative viral nucleic acid test for transplant patient management | QLX | 21 CFR 866.3183 | II |
1.3 Predicate Device
| Device Name | Predicate Device | 510(k) | Cleared |
|---|---|---|---|
| Alinity m EBV | Alinity m EBV | K212778 | 07/15/2022 |
1.4 Device Description
Alinity m EBV is an in vitro polymerase chain reaction (PCR) assay for the quantitation of EBV DNA in human plasma.
This device is similar to the predicate device originally cleared (K212778) with the exception that the subject device may use MomentaTaq DNA Polymerase as an
Page 7
alternative to KAPA2G DNA Polymerase in the reagent formulation of the assay. This formulation difference does not introduce any changes to sample processing, assay procedure, or data reduction.
Additional studies were initiated to support the formulation of the assay with MomentaTaq DNA Polymerase. Supplemental data from these studies were used with data previously obtained from the analytical and clinical testing studies submitted in K212778.
The steps of the Alinity m EBV consist of sample preparation, real-time PCR assembly, amplification/detection, result calculation, and reporting. All stages of the Alinity m EBV procedure are executed automatically by the Alinity m System. No intermediate processing or transfer steps are performed by the user. The Alinity m System is designed to be a random-access analyzer that can perform the Alinity m EBV assay in parallel with other Alinity m assays on the same instrument.
Alinity m EBV requires three separate assay specific kits as follows:
-
Alinity m EBV AMP Kit (List No. 09N43-095), consisting of 2 types of multi-well assay trays. The amplification trays (AMP TRAY 1) contain lyophilized, unit-dose PCR amplification/detection reagents and lyophilized, unit-dose IC in separate wells, and the activation trays (ACT TRAY 2) contain liquid unit-dose activation reagent. The intended storage condition for the Alinity m EBV AMP Kit is 2°C to 8°C.
-
Alinity m EBV CTRL Kit (List No. 09N43-085), consisting of negative controls, low positive controls, and high positive controls, each supplied as liquid in single-use tubes. The intended storage condition for the Alinity m EBV CTRL Kit is –25°C to –15°C.
-
Alinity m EBV CAL Kit (List No. 09N43-075), consisting of 2 calibrator levels, each supplied as liquid in single-use tubes. The intended storage condition for the Alinity m EBV CAL Kit is –25°C to –15°C.
Page 8
EBV DNA from human plasma is extracted automatically on-board in the Alinity m System using the Alinity m Sample Prep Kit 2, Alinity m Lysis Solution, and Alinity m Diluent Solution. The Alinity m System employs magnetic microparticle technology to facilitate nucleic acid capture, wash, and elution. The resulting purified nucleic acids are then combined with liquid unit-dose Alinity m EBV activation reagent and lyophilized unit-dose Alinity m EBV amplification/detection reagents and transferred into a reaction vessel. Alinity m Vapor Barrier Solution is then added to the reaction vessel which is then transferred to an amplification/detection unit for PCR amplification, and real-time fluorescence detection of EBV targets.
At the beginning of the Alinity m EBV sample preparation process, a lyophilized unit-dose IC on the AMP Tray is rehydrated by the Alinity m System and delivered into each sample preparation reaction. The IC is then processed through the entire sample preparation and real-time PCR procedure along with the specimens, calibrators, and controls to demonstrate proper sample processing and validity.
The Alinity m EBV amplification/detection reagents consist of enzymes, primers, probes, and activation reagents that enable polymerization and detection.
An EBV calibration curve is required for determination of EBV DNA concentration. Two levels of calibrators are processed through sample preparation and PCR to generate the calibration curve. The concentration of EBV DNA in specimens and controls is then calculated from the stored calibration curve.
Assay controls are tested at or above an established minimum frequency to help ensure that instrument and reagent performance remains satisfactory. During each control event, a negative control, a low-positive control, and a high-positive control are processed through sample preparation and PCR procedures that are identical to those used for specimens.
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The Alinity m EBV assay also utilizes the following:
- Alinity m EBV Application Specification File, (List No. 09N43-05B)
- Alinity m System and System Software (List No. 08N53-002)
- Alinity m Sample Prep Kit 2 (List No. 09N12-001)
- Alinity m Specimen Dilution Kit I (List No. 09N50-001)
- Alinity m System Solutions, (List No. 09N20):
- Alinity m Lysis Solution (List No. 09N20-001)
- Alinity m Diluent Solution (List No. 09N20-003)
- Alinity m Vapor Barrier Solution, (List No. 09N20-004)
- Alinity m Tubes and Caps (List No. 09N49):
- Alinity m LRV Tube (List No. 09N49-001)
- Alinity m Transport Tubes Pierceable Capped (List No. 09N49-010)
- Alinity m Transport Tube (List No. 09N49-011)
- Alinity m Pierceable Cap (List No. 09N49-012)
- Alinity m Aliquot Tube (List No. 09N49-013)
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1.5 Intended Use
Alinity m EBV is an in vitro polymerase chain reaction (PCR) assay for the quantitation of Epstein-Barr Virus (EBV) DNA in human EDTA plasma on the automated Alinity m System.
Alinity m EBV is intended for use as an aid in the management of EBV in transplant patients. In patients undergoing monitoring of EBV, serial DNA measurements can be used to indicate the need for potential treatment changes and to assess viral response to treatment.
The results from Alinity m EBV must be interpreted within the context of all relevant clinical and laboratory findings. Alinity m EBV is not intended to be used as a screening test for donors of blood, blood products, or human cells, tissues, and cellular and tissue-based products (HCT/Ps) for EBV.
1.6 Similarities and Differences to Predicate Device
The primary functional components of the Alinity m EBV assay are substantially equivalent to the current on-market Alinity m EBV assay (K212778), which is a legally marketed nucleic acid amplification test (NAAT) for the quantitative detection of EBV DNA.
The Alinity m EBV assay has the same intended use as the predicate device. The subject device and predicate device differ only in the DNA polymerase that may be used in manufacture of the assay (MomentaTaq vs. KAPA2G); this formulation difference does not raise new types of safety or effectiveness questions.
These devices are similar in that they are designed to prepare nucleic acids for amplification, amplify specific EBV DNA sequences, detect the amplified products, and report quantitative results.
The primary similarities and differences between the Alinity m EBV assay and the predicate device are shown in Table 1 and Table 2.
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Table 1. Similarities Between Alinity m EBV and Predicate Device
| Description | Subject Device | Predicate Device |
|---|---|---|
| Alinity m EBV | Alinity m EBV (K212778) | |
| Intended Use | Same | Alinity m EBV is an in vitro polymerase chain reaction (PCR) assay for the quantitation of Epstein-Barr Virus (EBV) DNA in human EDTA plasma on the automated Alinity m System. Alinity m EBV is intended for use as an aid in the management of EBV in transplant patients. In patients undergoing monitoring of EBV, serial DNA measurements can be used to indicate the need for potential treatment changes and to assess viral response to treatment. The results from Alinity m EBV must be interpreted within the context of all relevant clinical and laboratory findings. Alinity m EBV is not intended to be used as a screening test for donors of blood, blood products, or human cells, tissues, and cellular and tissue-based products (HCT/Ps) for EBV. |
| Assay Type | Same | Quantitative |
| Assay Targets | Same | 2 highly conserved regions of the EBV genome (gp350 and EBNA1) |
| Specimen Types | Same | EDTA Plasma |
| Sample Preparation Procedure | Same | Automated liquid handling and robotic manipulation platform |
| Amplification Technology | Same | Real-time polymerase chain reaction |
| Assay Controls | Same | • Negative Control• Low Positive Control• High Positive Control• Internal Control (IC) |
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Table 2. Differences Between Alinity m EBV and Predicate Device
| Feature | Current Application | Predicate Device |
|---|---|---|
| Alinity m EBV | Alinity m EBV (K212778) | |
| Amplification Enzymes | KAPA2G HotStart DNA Polymerase or MomentaTaq HotStart DNA Polymerase is the enzyme used for DNA amplification. | KAPA2G HotStart DNA Polymerase is the enzyme used for DNA amplification. |
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1.7 Performance Data
The following performance data were provided in support of safety, effectiveness, and substantial equivalence determination of the device.
1.7.1 Specific Performance Characteristics
The subject device is similar to the predicate device originally cleared (K212778) with the exception that the subject device may use MomentaTaq DNA Polymerase as an alternative to KAPA2G DNA Polymerase in the reagent formulation of the assay. Apart from the DNA polymerase enzyme used, the reagent formulation, including all oligonucleotide primers and probes (components and concentrations) is identical between the subject device and predicate device. There are no differences in the intended use, end-user workflow, instrument workflow, assay software, reagent manufacturing, or final product release specifications.
Additional analytical performance studies listed in Table 3 were conducted to confirm the ability of the Alinity m EBV assay formulated with MomentaTaq DNA Polymerase (subject device) to meet the performance claims established in the corresponding studies submitted in K212778 for the Alinity m assay formulated with KAPA2G DNA Polymerase (predicate device).
Table 3. Additional Supporting Studies (MomentaTaq formulation)
| Study Description / Performance Characteristic | 510(k) Summary Section |
|---|---|
| Limit of Detection | 1.7.1.1 |
| Linear Range | 1.7.1.2 |
| Precision | 1.7.1.3 |
| Lower Limit of Quantitation | 1.7.1.4 |
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All other specific performance characteristics of the subject device are supported by the studies submitted in K212778 (original submission and Amendments 1 and 2). Refer to Table 4.
Table 4. Supporting Studies Submitted in K212778
| Study Description / Performance Characteristic | Reference |
|---|---|
| Traceability to the WHO Standard | Original submission |
| Limit of Detection for EBV Type 2 | Original submission |
| Linearity for EBV Type 2 | Amendment 2 |
| Analytical Specificity – Potential Cross-Reactants | Original submission |
| Analytical Specificity – Potentially Interfering Substances | Original submission |
| Carryover | Amendment 1 |
| Alinity m EBV Testing Using Dilution Procedure | Original submission |
| Precision of Alinity m EBV Using Dilution Procedure | Original submission |
| Confirmation of LLoQ Using Dilution Procedure | Original submission |
1.7.1.1 Limit of Detection
A limit of detection (LoD) study was conducted previously to support the clearance of K212778. Please refer to the decision summary for K212778.
An additional study was conducted to confirm the LoD claim of Alinity m EBV (20 IU/mL) for the MomentaTaq formulation of the assay.
LoD for EBV type 1 was evaluated by testing dilutions of the 1st World Health Organization (WHO) International Standard for Epstein-Barr Virus for Nucleic Acid Amplification Techniques (NIBSC code 09/260) prepared in EBV negative human plasma. Testing for each EBV DNA concentration was performed with 3 lots of amplification reagents across multiple days. The data from the study demonstrated that the Alinity m EBV assay formulated with MomentaTaq DNA Polymerase had an overall detection rate of 97.2% at 20 IU/mL for all reagent lots combined with a 95% Confidence Interval of (93.0%, 98.9%). Refer to Table 5.
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Table 5. Alinity m EBV (MomentaTaq formulation) Limit of Detection (LoD) Results
| Panel | Target Concentration (IU/mL) | AMP Kit Reagent Lot | Total No. of Valid Replicates | No. of Replicates Detected | Detection Rate (%) | 95% Score Confidence Interval (%) |
|---|---|---|---|---|---|---|
| 1 | 15 | 1 | 48 | 45 | 93.8 | (83.2, 97.9) |
| 2 | 48 | 46 | 95.8 | (86.0, 98.8) | ||
| 3 | 47 | 45 | 95.7 | (85.8, 98.8) | ||
| All Combined | 143 | 136 | 95.1 | (90.2, 97.6) | ||
| 2 | 20ᵃ | 1 | 47 | 47 | 100.0 | (92.4, 100.0) |
| 2 | 48 | 46 | 95.8 | (86.0, 98.8) | ||
| 3 | 47 | 45 | 95.7 | (85.8, 98.8) | ||
| All Combined | 142 | 138 | 97.2 | (93.0, 98.9) | ||
| 3 | 50 | 1 | 48 | 48 | 100.0 | (92.6, 100.0) |
| 2 | 48 | 48 | 100.0 | (92.6, 100.0) | ||
| 3 | 47 | 47 | 100.0 | (92.4, 100.0) | ||
| All Combined | 143 | 143 | 100.0 | (97.4, 100.0) | ||
| 4 | 100 | 1 | 48 | 48 | 100.0 | (92.6, 100.0) |
| 2 | 47 | 47 | 100.0 | (92.4, 100.0) | ||
| 3 | 47 | 47 | 100.0 | (92.4, 100.0) | ||
| All Combined | 142 | 142 | 100.0 | (97.4, 100.0) |
ᵃ Alinity m EBV assay limit of detection (LoD) claim
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1.7.1.2 Linear Range
A linearity study was conducted previously to support the clearance of K212778. Please refer to the decision summary for K212778.
An additional study was conducted to confirm that the Alinity m EBV assay formulated with MomentaTaq was linear across the claimed quantitation range (50 IU/mL [1.70 Log IU/mL] to 200,000,000 IU/mL [8.30 Log IU/mL]).
Linearity was evaluated by testing 16 panel members that spanned the intended quantitation range of the assay (50 IU/mL to 200,000,000 IU/mL), including a panel member below the expected Lower Limit of Quantification (LLoQ) at 15 IU/mL and a panel member exceeding the expected Upper Limit of Quantification (ULoQ) at 250,000,000 IU/mL. Panels prepared with different specimen types were designed to have at least a 2 Log IU/mL overlap with adjacent sample types. Refer to Table 6.
Table 6. Panel Members for Linearity
| Panel Member | Targeted EBV Concentration | Source | |
|---|---|---|---|
| IU/mL | Log IU/mL | ||
| 01 | 250,000,000 | 8.40 | Plasmid DNA |
| 02 | 10,000,000 | 7.00 | Plasmid DNA |
| 03 | 1,000,000 | 6.00 | Plasmid DNA |
| 04 | 130,000 | 5.11 | Plasmid DNA |
| 05 | 100,000 | 5.00 | Cultured virus |
| 06 | 20,000 | 4.30 | Plasmid DNA |
| 07 | 10,000 | 4.00 | Cultured virus |
| 08 | 5,000 | 3.70 | Plasmid DNA |
| 09 | 2,500 | 3.40 | Clinical specimen |
| 10 | 1,000 | 3.00 | Plasmid DNA |
| 11 | 500 | 2.70 | Cultured virus |
| 12 | 200 | 2.30 | Clinical specimen |
| 13 | 50 | 1.70 | Clinical specimen |
| 14 | 25 | 1.40 | Plasmid DNA |
| 15 | 20 | 1.30 | Cultured virus |
| 16 | 15 | 1.18 | Clinical specimen |
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The data from the study demonstrated that the Alinity m EBV assay formulated with MomentaTaq DNA Polymerase was linear across the quantitation range from 15 IU/mL to 250,000,000 IU/mL (1.18 Log IU/mL to 8.40 Log IU/mL). Representative results for Alinity m EBV linearity performance are shown in Figure 1.
Figure 1. Alinity m EBV (MomentaTaq formulation) Linearity
[THIS IS FIGURE: A scatter plot showing linearity data with Expected Concentration (Log IU/mL) on x-axis (0-9) and Alinity m EBV Ct (Log IU/mL) on y-axis (0-9). The plot shows a strong linear relationship with r = 1.000 and data points following a diagonal line from approximately (1,1) to (8.5,8.5).]
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1.7.1.3 Precision
A precision study was conducted previously to support the clearance of K212778. Please refer to the decision summary for K212778.
An additional study was conducted to confirm the precision claim of Alinity m EBV (as follows) for the MomentaTaq formulation of the assay:
-
Within-laboratory standard deviation (SD) of ≤ 0.25 Log IU/mL for plasma samples containing 500 IU/mL to 200,000,000 IU/mL (2.70 Log IU/mL to 8.30 Log IU/mL) EBV DNA
-
Within-laboratory SD of ≤ 0.50 Log IU/mL for plasma samples containing 20 IU/mL to 500 IU/mL (1.30 Log IU/mL to 2.70 Log IU/mL) EBV DNA
Precision was evaluated by testing 8 panel members with EBV concentrations ranging from 20 IU/mL to 2 × 10⁸ IU/mL (1.30 Log IU/mL to 8.30 Log IU/mL). Panel members were prepared by diluting EBV-positive clinical specimen, EBV cultured virus or synthetic DNA in human plasma. EBV synthetic DNA was only used for panel members with high target concentrations (> 6 Log IU/mL). The quantitation of the panel members was traceable to the 1st WHO International Standard for Epstein Barr virus (EBV) (NIBSC code 09/260).
Three lots of Alinity m EBV amplification reagents manufactured with MomentaTaq DNA Polymerase were run on 1 Alinity m System by 3 operators (each of the 3 operators used 1 lot of amplification reagents). Three replicates of each panel member were run twice each day for 15 days for each lot/operator combination with a minimum separation of 2 hours between successive runs of a panel member.
The overall precision analysis summary for the Alinity m EBV assay formulated with MomentaTaq DNA Polymerase is presented in Table 7.
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Table 7. Alinity m EBV (MomentaTaq formulation) Precision Analysis – All Reagent Lots
| Panel | N | Mean Concentration (Log IU/mL) | Within-Run Component | Between-Run Component | Between-Day Component | Within-Laboratoryᵃ | Between-Lot Component | Totalᵇ |
|---|---|---|---|---|---|---|---|---|
| SD | %CV | SD | %CV | SD | %CV | |||
| 01 | 270 | 1.35 | 0.24 | 18.0 | 0.11 | 8.2 | 0.00 | 0.0 |
| 02 | 270 | 1.69 | 0.19 | 11.1 | 0.00 | 0.0 | 0.06 | 3.3 |
| 03 | 270 | 2.78 | 0.08 | 2.9 | 0.00 | 0.0 | 0.02 | 0.7 |
| 04 | 270 | 3.99 | 0.07 | 1.6 | 0.02 | 0.4 | 0.00 | 0.0 |
| 05 | 270 | 5.07 | 0.05 | 1.0 | 0.03 | 0.6 | 0.01 | 0.2 |
| 06 | 270 | 6.10 | 0.05 | 0.9 | 0.03 | 0.5 | 0.00 | 0.0 |
| 07 | 270 | 7.03 | 0.06 | 0.9 | 0.02 | 0.3 | 0.01 | 0.1 |
| 08 | 270 | 8.39 | 0.17 | 2.0 | 0.08 | 1.0 | 0.00 | 0.0 |
ᵃ Within-Laboratory includes Within-Run, Between-Run, and Between-Day Components.
ᵇ Total includes Within-Run, Between-Run, Between-Day, and Between-Lot Components.
The results demonstrated that the Alinity m EBV assay has a within-laboratory SD of 0.25 Log IU/mL or less for EBV DNA panels targeted to 500 IU/mL to 200,000,000 IU/mL (2.70 Log IU/mL to 8.30 Log IU/mL) and a within-laboratory SD of 0.50 Log IU/mL or less for EBV DNA panels targeted to 20 IU/mL to < 500 IU/mL (1.30 Log IU/mL to < 2.70 Log IU/mL).
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Table 8. Equivalence Evaluation of Total SD and Total %CV (Newᵃ vs. Originalᵇ)
| Panel Member | Mean Concentration | Total SD | Total %CV | Ratios and 95% CI | Clinically Acceptable?ᵉ |
|---|---|---|---|---|---|
| New (Log IU/mL) | Original (Log IU/mL) | New SD | Original SD | New % CV | |
| 01 | 1.35 | 1.43 | 0.27 | 0.26 | 19.8 |
| 02ᶜ | 1.69 | 2.2 | 0.20 | 0.2 | 11.6 |
| 03 | 2.78 | 2.77 | 0.08 | 0.13 | 3.0 |
| 04 | 3.99 | 4.06 | 0.07 | 0.08 | 1.7 |
| 05 | 5.07 | 5.05 | 0.06 | 0.07 | 1.2 |
| 06 | 6.10 | 6.09 | 0.06 | 0.07 | 1.0 |
| 07 | 7.03 | 7.05 | 0.07 | 0.06 | 1.0 |
| 08ᵈ | 8.39 | 8.22 | 0.19 | 0.08 | 2.3 |
ᵃ "New" Mean Concentration, Total SD, and Total %CV data for Alinity m EBV assay formulated with MomentaTaq DNA Polymerase
ᵇ "Original" Mean Concentration, Total SD, and Total %CV data for on-market Alinity m EBV assay formulated with KAPA2G DNA Polymerase (submitted in K212778)
ᶜ Panel 02 in the New precision study targeting 1.7 Log IU/mL is compared to panel 02 in the Original precision study targeting 2.0 Log IU/mL.
ᵈ Panel 08 in the New precision study targeting 8.3 Log IU/mL is compared to panel 09 in the Original precision study targeting 8.3 Log IU/mL. NOTE: Panel 08 in the Original precision study targeting 8.0 Log IU/mL is not analyzed for comparison.
ᵉ Precision results were effectively equivalent if the 95% confidence interval for the ratio of SD or ratio of %CVs between the MomentaTaq DNA Polymerase ("New") and KAPA2G DNA Polymerase ("Original") contained 1.00 or the upper bound of the 95% confidence interval for the ratio of SD or ratio of %CV is < 1.00; these results were therefore "Clinically Acceptable."
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1.7.1.4 Lower Limit of Quantitation (LLoQ)
A lower limit of quantitation (LLoQ) study was conducted previously to support the clearance of K212778. Please refer to the decision summary for K212778.
An additional study was conducted to confirm the LLoQ claim of Alinity m EBV (50 IU/mL) for the MomentaTaq formulation of the assay.
The LLoQ is defined as the lowest concentration at which EBV DNA is reliably quantitated within an acceptable total error. Total error was estimated for detected samples from the LoD study (Section 1.7.1.1) by 2 methods:
- Total Analytical Error (TAE) = |bias| + 2 × SD, and
- Total Error (TE) = SQRT(2) × 2 × SD
The results of the calculations are shown in Table 9.
Panel members were dilutions of the 1st WHO International Standard for Epstein-Barr Virus for Nucleic Acid Amplification Techniques (NIBSC code 09/260) prepared in EBV-negative plasma.
The results of these analyses support a claimed LLoQ of 50 IU/mL (1.70 Log IU/mL), with an acceptable level of accuracy and precision (ie, TAE and TE less than or equal to 1.00 Log IU/mL).
Table 9. Alinity m EBV (MomentaTaq formulation) TAE and TE
| Target Concentration (Log IU/mL) | Mean Concentration (Log IU/mL) | Biasᵃ (Log IU/mL) | SD (Log IU/mL) | TAE (Log IU/mL) | TE (Log IU/mL) |
|---|---|---|---|---|---|
| 1.18 | 0.96 | –0.22 | 0.32 | 0.86 | 0.91 |
| 1.30 | 1.07 | –0.23 | 0.33 | 0.89 | 0.93 |
| 1.70 | 1.52 | –0.18 | 0.20 | 0.59 | 0.57 |
| 2.00 | 1.86 | –0.14 | 0.17 | 0.48 | 0.47 |
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1.7.1 Clinical Performance Characteristics
1.7.1.1 Clinical Reproducibility
A reproducibility study was conducted previously to support the clearance of K212778. Please refer to the decision summary for K212778.
An additional study was conducted to evaluate the reproducibility of the Alinity m EBV assay formulated with MomentaTaq DNA Polymerase.
Reproducibility performance of Alinity m EBV was evaluated by testing a 9-member reproducibility panel, which included 8 positive panel members and 1 negative panel member. The positive panel members were prepared using EBV positive clinical specimen, cultured virus, or plasmid DNA diluted in human EDTA plasma. The concentration levels spanned the quantitation range of the assay.
One lot each of Alinity m EBV AMP Kit, Alinity m EBV CAL Kit, Alinity m EBV CTRL Kit, and Alinity m Sample Prep Kit 2 lots were used. Testing was performed at 3 clinical sites on 5 non-consecutive days with 2 runs per day. Four replicates of each panel member were tested on each of 5 days to ensure a minimum of 3 valid replicates for analysis.
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The reproducibility results are summarized in Table 10 (for the positive panel members) and in Table 11 (for the negative panel member).
Table 10. Alinity m EBV (MomentaTaq formulation) Reproducibility for Positive Panel Members
| Panel | N | Mean Concentration (Log IU/mL) | Within-Run Component | Between-Run Component | Between-Day Component | Within-Laboratoryᵃ | Between-Site Component | Totalᵇ |
|---|---|---|---|---|---|---|---|---|
| SD | %CV | SD | %CV | SD | %CV | |||
| 1 | 118 | 7.88 | 0.06 | 0.7 | 0.01 | 0.1 | 0.00 | 0.0 |
| 2 | 116 | 6.82 | 0.04 | 0.6 | 0.00 | 0.0 | 0.02 | 0.3 |
| 3 | 120 | 6.00 | 0.05 | 0.8 | 0.02 | 0.4 | 0.01 | 0.2 |
| 4 | 118 | 5.02 | 0.04 | 0.8 | 0.02 | 0.3 | 0.02 | 0.4 |
| 5 | 120 | 4.00 | 0.05 | 1.2 | 0.00 | 0.1 | 0.01 | 0.3 |
| 6 | 118 | 3.09 | 0.07 | 2.4 | 0.01 | 0.2 | 0.00 | 0.0 |
| 7 | 117 | 1.74 | 0.19 | 10.9 | 0.00 | 0.0 | 0.00 | 0.0 |
| 8 | 113 | 1.39 | 0.29 | 21.2 | 0.04 | 2.5 | 0.00 | 0.0 |
ᵃ Within-Laboratory includes Within-Run, Between-Run, and Between-Day Variance Components.
ᵇ Total (Overall) includes Within-Run, Between-Run, Between-Day, and Between-Site Variance Components.
Table 11. Alinity m EBV (MomentaTaq formulation) Reproducibility for Negative Panel Member
| Panel | Site | N | Positive | Negative | Negative Agreement Rate | 95% CI |
|---|---|---|---|---|---|---|
| 9 | All | 119 | 1 | 118 | 99.2% | (95.4%, 99.9%) |
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The Total SD and Total %CV for each panel member in this study were compared with the Total SD and Total %CV from the original reproducibility study submitted in K212778. Refer to Table 12.
Table 12. Equivalence Evaluation of Total SD and Total %CV (Newᵃ vs. Originalᵇ)
| Panel | Mean Concentration | Total SD | Total %CV | Ratios and 95% CI | Clinically Acceptable?ᶜ |
|---|---|---|---|---|---|
| New (Log IU/mL) | Original (Log IU/mL) | New SD | Original SD | New %CV | |
| 1 | 7.88 | 8.40 | 0.06 | 0.19 | 0.7 |
| 2 | 6.82 | 7.04 | 0.04 | 0.14 | 0.6 |
| 3 | 6.00 | 5.76 | 0.06 | 0.10 | 1.0 |
| 4 | 5.02 | 5.04 | 0.06 | 0.12 | 1.2 |
| 5 | 4.00 | 4.01 | 0.06 | 0.09 | 1.6 |
| 6 | 3.09 | 3.07 | 0.08 | 0.09 | 2.6 |
| 7 | 1.74 | 1.64 | 0.19 | 0.23 | 11.1 |
| 8 | 1.39 | 1.20 | 0.30 | 0.30 | 21.3 |
ᵃ "New" Mean Concentration, Total SD, and Total %CV data for Alinity m EBV assay formulated with MomentaTaq DNA Polymerase
ᵇ "Original" Mean Concentration, Total SD, and Total %CV data for on-market Alinity m EBV assay formulated with KAPA2G DNA Polymerase (submitted in K212778)
ᶜ Reproducibility results were effectively equivalent if the 95% confidence interval for the ratio of SD or ratio of %CVs between the MomentaTaq DNA Polymerase ("New") and KAPA2G DNA Polymerase ("Original") contained 1.00 or the ratio of SD or ratio of %CV is <1.00; these results were therefore considered "Clinically Acceptable."
1.7.1.2 Clinical Performance
A method comparison study was conducted with the Alinity m EBV assay formulated with MomentaTaq DNA Polymerase to demonstrate equivalence to the FDA-cleared (on-market) Alinity m EBV assay formulated with KAPA2G DNA Polymerase (K212778)
Regression and bias analysis included a total of 124 samples with results that were within the common quantitation range of both the Alinity m EBV assay formulated with MomentaTaq (Alinity m EBV IUO) and the comparator assay (on-market Alinity m EBV). Figure 2 shows the results of the Deming regression analysis with a slope of 1.00, an intercept of 0.01, and a correlation coefficient of 0.993. The mean bias between Alinity m EBV IUO and the comparator (Alinity m EBV IUO minus comparator) was –0.01 Log IU/mL with a 95% Confidence Interval of (–0.03, –0.01).
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Figure 2. Deming Regression Analysis
y = 1.00x + 0.01
r = 0.993
n = 124
[THIS IS FIGURE: A scatter plot showing Deming regression analysis with Alinity m EBV IUO Quantitation (Log IU/mL) on y-axis (1-8) and On-market Alinity m EBV Quantitation (Log IU/mL) on x-axis (1-8). The plot shows a strong linear relationship with data points closely following the regression line.]
Systematic difference between Alinity m EBV assay formulated with MomentaTaq and the on-market Alinity m EBV assay (comparator) at 4 selected viral load levels is shown in Table 13.
Table 13. Systematic Difference at Selected Viral Load Levels
| Target Viral Load Level (based on comparator) (Log IU/mL) | Systematic Difference (Log IU/mL) |
|---|---|
| 1.70 (LLoQ) | 0.02 |
| 3.00 | 0.01 |
| 4.00 | 0.01 |
| 5.00 | 0.00 |
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1.8 Conclusions Drawn from the Studies
The additional analytical and clinical study results demonstrate that the Alinity m EBV assay formulated with MomentaTaq DNA Polymerase performs comparably to the FDA-cleared Alinity m EBV assay formulated with KAPA2G DNA Polymerase.
The results support a substantial equivalence decision for Alinity m EBV formulated with MomentaTaq DNA Polymerase.
§ 866.3183 Quantitative viral nucleic acid test for transplant patient management.
(a)
Identification. A quantitative viral nucleic acid test for transplant patient management is identified as a device intended for prescription use in the detection of viral pathogens by measurement of viral DNA or RNA using specified specimen processing, amplification, and detection instrumentation. The test is intended for use as an aid in the management of transplant patients with active viral infection or at risk for developing viral infections. The test results are intended to be interpreted by qualified healthcare professionals in conjunction with other relevant clinical and laboratory findings.(b)
Classification. Class II (special controls). The special controls for this device are:(1) The labeling required under § 809.10(b) of this chapter must include:
(i) A prominent statement that the device is not intended for use as a donor screening test for the presence of viral nucleic acid in blood or blood products.
(ii) Limitations which must be updated to reflect current clinical practice. These limitations must include, but are not limited to, statements that indicate:
(A) Test results are to be interpreted by qualified licensed healthcare professionals in conjunction with clinical signs and symptoms and other relevant laboratory results; and
(B) Negative test results do not preclude viral infection or tissue invasive viral disease and that test results must not be the sole basis for patient management decisions.
(iii) A detailed explanation of the interpretation of results and acceptance criteria must be provided and include specific warnings regarding the potential for variability in viral load measurement when samples are measured by different devices. Warnings must include the following statement, where applicable: “Due to the potential for variability in [analyte] measurements across different [analyte] assays, it is recommended that the same device be used for the quantitation of [analyte] when managing individual patients.”
(iv) A detailed explanation of the principles of operation and procedures for assay performance.
(2) Design verification and validation must include the following:
(i) Detailed documentation of the device description, including all parts that make up the device, ancillary reagents required for use with the assay but not provided, an explanation of the methodology, design of the primer/probe sequences, rationale for the selected gene target, and specifications for amplicon size, guanine-cytosine content, and degree of nucleic acid sequence conservation. The design and nature of all primary, secondary and tertiary quantitation standards used for calibration must also be described.
(ii) A detailed description of the impact of any software, including software applications and hardware-based devices that incorporate software, on the device's functions;
(iii) Documentation and characterization (
e.g., determination of the identity, supplier, purity, and stability) of all critical reagents and protocols for maintaining product integrity throughout its labeled shelf-life.(iv) Stability data for reagents provided with the device and indicated specimen types, in addition to the basis for the stability acceptance criteria at all time points chosen across the spectrum of the device's indicated life cycle, which must include a time point at the end of shelf life.
(v) All stability protocols, including acceptance criteria.
(vi) Final lot release criteria along with documentation of an appropriate justification 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) Risk analysis and documentation demonstrating how risk control measures are implemented to address device system hazards, such as Failure Mode 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 viral stains (
e.g., regular review of published literature and annual in silico analysis of target sequences to detect possible primer or probe mismatches). All results of this protocol, including any findings, must be documented.(viii) Analytical performance testing that includes:
(A) Detailed documentation of the following analytical performance studies: limit of detection, upper and lower limits of quantitation, inclusivity, precision, reproducibility, interference, cross reactivity, carry-over, quality control, specimen stability studies, and additional studies as applicable to specimen type and intended use for the device;
(B) Identification of the viral strains selected for use in analytical studies, which must be representative of clinically relevant circulating strains;
(C) Inclusivity study results obtained with a variety of viral genotypes as applicable to the specific assay target and supplemented by in silico analysis;
(D) Reproducibility studies that include the testing of three independent production lots;
(E) Documentation of calibration to a reference standard that FDA has determined is appropriate for the quantification of viral DNA or RNA (
e.g., a recognized consensus standard); and(F) Documentation of traceability performed each time a new lot of the standardized reference material to which the device is traceable is released, or when the field transitions to a new standardized reference material.
(ix) Clinical performance testing that includes:
(A) Detailed documentation from either a method comparison study with a comparator that FDA has determined is appropriate, or results from a prospective clinical study demonstrating clinical validity of the device;
(B) Data from patient samples, with an acceptable number of the virus-positive samples containing an analyte concentration near the lower limit of quantitation and any clinically relevant decision points. If an acceptable number of virus-positive samples containing an analyte concentration near the lower limit of quantitation and any clinically relevant decision cannot be obtained, contrived samples may be used to supplement sample numbers when appropriate, as determined by FDA;
(C) The method comparison study must include predefined maximum acceptable differences between the test and comparator method across all primary outcome measures in the clinical study protocol; and
(D) The final release test results for each lot used in the clinical study.