(317 days)
No
The description focuses on standard PCR assay procedures, automated sample processing, and quantitative analysis based on a calibration curve. There is no mention of AI/ML algorithms for data interpretation, pattern recognition, or result calculation beyond standard quantitative methods.
No.
The device is an in vitro diagnostic (IVD) PCR assay intended for the quantitation of EBV DNA to aid in the management of EBV in transplant patients by providing serial DNA measurements to indicate the need for potential treatment changes and to assess viral response. It performs diagnostic measurements, it does not provide therapy itself.
Yes
The text explicitly states, "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" and "Alinity m EBV is intended for use as an aid in the management of EBV in transplant 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." This indicates its use in identifying and quantifying a biological marker (EBV DNA) to assist in patient management, which is a diagnostic purpose.
No
The device is an in vitro diagnostic (IVD) assay that requires specific reagent kits, controls, calibrators, and is performed on a dedicated hardware system (Alinity m System). It is not solely software.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The "Intended Use / Indications for Use" section explicitly states that the Alinity m EBV is an in vitro polymerase chain reaction (PCR) assay. It is intended for use as an aid in the management of EBV in transplant patients by quantitating EBV DNA in human EDTA plasma. This clearly indicates that the device is used to test samples taken from the human body outside of the body to provide diagnostic information.
- Device Description: The "Device Description" further reinforces this by describing the assay as an in vitro PCR assay for the quantitation of EBV DNA in human plasma. It details the process of sample preparation, amplification, detection, and result calculation, all of which are performed on biological samples in vitro.
- Components: The description of the assay-specific kits (AMP Kit, CTRL Kit, CAL Kit) and other accessories (Sample Prep Kit, Lysis Solution, Diluent Solution, etc.) are all components used in the laboratory setting to perform the diagnostic test on the patient sample.
- Performance Studies: The "Summary of Performance Studies" describes analytical and clinical performance evaluations conducted on human plasma samples, which is characteristic of an IVD.
The definition of an In Vitro Diagnostic (IVD) is a medical device that is intended for use in the examination of specimens derived from the human body to provide information for diagnostic, monitoring, or compatibility purposes. The Alinity m EBV fits this definition perfectly.
N/A
Intended Use / Indications for 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 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 products, or human cells, tissues, and cellular and tissue-based products (HCT/Ps) for EBV.
Product codes (comma separated list FDA assigned to the subject device)
QLX
Device Description
Alinity m EBV is an in vitro polymerase chain reaction (PCR) assay for the quantitation of EBV DNA in human plasma.
The steps of the Alinity m EBV assay 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 multi-well amplification trays (AMP Trays) containing lyophilized, unit-dose PCR amplification/detection reagents and multi-well activation trays (ACT Trays) containing liquid, unit-dose activation reagents (MgCl2, TMAC, KCl, and ProClin). 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 a negative control, a low positive control, and a high positive control, each supplied as liquid in single-use tubes. The intended storage condition for the Alinity m EBV CTRL Kit is —15°C to —25°C.
- . Alinity m EBV CAL Kit (List No. 09N43-075) consisting of two levels of calibrators (CAL A and CAL B), each supplied as liquid in single-use tubes. The intended storage condition for the Alinity m EBV CAL Kit is -15°C to -25°C.
EBV DNA from specimens 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 the 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.
An EBV calibration curve is required for the quantitation of EBV targets. Two levels of calibrators are processed through sample preparation and real-time 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 remain satisfactory. During each control event, a negative control, a low-positive control, and a high-positive control are processed through sample preparation and real-time PCR procedures that are identical to those used for specimens.
At the beginning of the Alinity m EBV sample preparation process, a lyophilized unit -dose of Internal Control on the AMP Tray is rehydrated by the Alinity m System and delivered into each sample preparation reaction. The Internal Control 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 assay validity.
The Alinity m EBV amplification and detection reagents include primers and probes that amplify and detect dual targets in the EBV genome. Amplification and detection of the two EBV targets ensures sensitive detection of the viral genome even at low levels.
The Alinity m EBV assay also utilizes the following accessories:
- . Alinity m EBV Application Specification File, List No. 09N43-05A
- . 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 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
- . 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
Mentions image processing
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Mentions AI, DNN, or ML
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Input Imaging Modality
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Anatomical Site
Not Found
Indicated Patient Age Range
Not Found
Intended User / Care Setting
Not Found
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
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Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
- Traceability to the WHO Standard: Primary calibrators and assay product calibrators with known concentrations were used throughout product development and product manufacturing to establish traceability to the 1st World Health Organization (WHO) International Standard for Epstein-Barr virus for Nucleic Acid Amplification Techniques (NIBSC code: 09/260). The concentrations tested for the WHO standard were 3.00 Log IU/mL and 5.00 Log IU/mL. The target concentrations tested for the primary calibrators were 3.00 Log IU/mL and 7.00 Log IU/mL. The Alinity m EBV product calibrators and controls were also tested along with the primary calibrators and the WHO standard. All of the tested material had observed EBV concentrations similar to the target concentrations, and were linear across the assay's quantitation range.
- Limit of Detection (LoD) for EBV type 1: Testing was performed with 4 lots of amplification reagents across multiple days. Probit analysis of the data using the least sensitive lot (Lot 3) determined that the concentration of EBV DNA in plasma detected with 95% probability (LoD by Probit) was 19.56 IU/mL with a 95% confidence interval (CI) of (13.09 IU/mL, 39.39 IU/mL). The claimed LoD of Alinity m EBV is 20 IU/mL (1.30 Log IU/mL) in plasma.
- Limit of Detection for EBV Type 2: Cultured virus for EBV type 2 was diluted to 3 different concentrations in EBV negative human plasma. Testing was performed using one lot of amplification reagents across multiple days. Alinity m EBV detected 95% or greater of EBV samples at and above 15 IU/mL (1.18 Log IU/mL) in plasma. These results demonstrate the ability of Alinity m EBV to detect EBV type 2 at the claimed LoD.
- Linear Range (EBV type 1 and type 2): The quantitation range of Alinity m EBV is from the lower limit of quantitation (LLQ) of 50 IU/mL (1.70 Log IU/mL) to the upper limit of quantitation (ULoQ) of 200,000,000 IU/mL (8.30 Log IU/mL). Linearity was assessed by testing a dilution series of EBV type 1 and type 2 in negative human plasma, consisting of 16 panel levels targeted in the range of 10 IU/mL to 400,000,000 IU/mL (1.00 Log IU/mL to 8.60 Log IU/mL). Panel quantitation values were traceable to the 1st World Health Organization (WHO) International Standard for Epstein-Barr Virus for Nucleic Acid Amplification Techniques (NIBSC code: 09/260). Alinity m EBV was linear across the quantitation range from 50 IU/mL to 200,000,000 IU/mL (1.70 Log IU/mL to 8.30 Log IU/mL) for both EBV type 1 and type 2.
- Precision: Precision was determined by analyzing a 9-member plasma panel, tested in 4 replicates, twice each day for 12 days, on 3 Alinity m Systems operated by 3 operators, using 3 AMP kit lots, for a total of 288 replicates per panel member. The within-laboratory standard deviation (SD) was less than or equal to 0.25 Log IU/mL for EBV DNA panels targeted in the range of 2.70 Log IU/mL to 8.30 Log IU/mL (500 IU/mL to 200.000.000 IU/mL), and less than or equal to 0.50 Log IU/mL for EBV DNA panels targeted in the range of 1.30 Log IU/mL to less than 2.70 Log IU/mL (20 IU/mL to less than 500 IU/mL).
- Lower Limit of Quantitation (LLoQ): 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 by two methods: Total Analytical Error (TAE) and Total Error (TE). The results support a claimed LLoQ of 50.00 IU/mL (1.70 Log IU/mL) for Alinity m EBV.
- Analytical Specificity – Potential Cross-Reactants: The analytical specificity was evaluated with a panel of microorganisms (viruses, bacteria, fungi) in EBV negative plasma, positive plasma targeted to 60 IU/mL EBV DNA, and positive plasma targeted to 10,000 IU/mL EBV DNA. Microorganisms were tested at a final concentration of 10^5 Units/mL for viruses and fungi or 10^6 Units/mL for bacteria. No cross-reactivity or interference was observed.
- Analytical Specificity - Potentially Interfering Substances: The effects of endogenous substances (albumin, hemoglobin, triglycerides, bilirubin, human genomic DNA) and high levels of therapeutic drugs commonly prescribed in transplant patients were evaluated by testing 8 negative samples, 8 positive samples targeted to 60 IU/mL and 8 positive samples targeted to 10,000 IU/mL EBV DNA. No interference was observed.
- Carryover: The carryover rate for Alinity m EBV was determined by analyzing 648 valid replicates of EBV negative samples processed from alternating positions with 647 valid replicates of high concentrated EBV positive samples greater than or equal to 20,000,000 IU/mL, across a minimum of 27 runs. The carryover resulting in a detectable concentration greater than or equal to LoD was 0.3% (95% CI: 0.1% to 1.1%). The carryover resulting in EBV detection was 1.2% (95% CI: 0.6% to 2.4%).
- Alinity m EBV Testing Using Dilution Procedure: The 1:2.5 dilution procedure was evaluated by comparing quantitation of neat samples and samples tested using the Alinity m EBV dilution procedure for five plasma panel members. The differences in mean ranged from -0.01 Log IU/mL to 0.23 Log IU/mL.
- Precision of Alinity m EBV Using Dilution Procedure: Precision was determined by analyzing 3 plasma panel members tested in at least 3 replicates, twice each day for 12 days, on 3 Alinity m Systems with 3 Alinity m Specimen Dilution Kit I lots and 3 Alinity m EBV AMP Kit lots by 3 operators, for a total of at least 216 replicates.
- Confirmation of the LLoQ Using Dilution Procedure: Confirmation testing for Alinity m EBV LLoQ using the dilution procedure was performed by testing 2 panel members at 50 IU/mL (LLoQ) and 60 IU/mL (near LLoQ) with a dilution factor of 1:2.5. A minimum of 14 replicates per day of each panel level were tested using the dilution procedure in 3 runs across 3 days. The accuracy and precision at 20 IU/mL were confirmed for Alinity m EBV testing using the 1:2.5 dilution procedure.
- Clinical Reproducibility: Reproducibility performance was evaluated by testing a 9-member reproducibility panel (8 positive, 1 negative). A total of 3 Alinity m EBV AMP Kit lots, 3 Alinity m EBV CAL Kit lots, 3 Alinity m EBV CTRL Kit lots and 3 Alinity m Sample Prep Kit 2 lots were used. Three clinical sites each tested 2 unique reagent lot combinations on 5 non-consecutive days. Six replicates of each panel member were tested on each of 5 days.
- Clinical Performance: Alinity m EBV results were compared to those of an FDA-cleared EBV nucleic acid test using 558 EDTA plasma samples (542 clinical specimens from HSCT and SOT subjects, and 16 contrived samples). Testing was performed at 3 clinical testing sites with 3 Alinity m EBV reagent kit lots. Of 550 valid samples, 388 were detected by Alinity m EBV and 162 were not. For a subset of 44 confirmed EBV DNA negative clinical specimens, the NPA with the Comparator assay was 100.0% (44/44) with a 95% confidence interval of (92.0%, 100.0%).
- Regression and bias analysis: Included 239 samples with results that were within the common quantitation range of both Alinity m EBV and the comparator assay. Deming regression analysis showed a slope of 1.01, intercept of 0.05, and correlation coefficient of 0.974. The mean bias between Alinity m EBV and the comparator (Alinity m EBV minus comparator) was 0.09 Log IU/mL with a 95% CI of (0.06, 0.12).
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
- LoD (EBV type 1): 19.56 IU/mL detected with 95% probability. Claimed LoD: 20 IU/mL (1.30 Log IU/mL).
- LoD (EBV type 2): ≥ 95% detection rate at 15 IU/mL (1.18 Log IU/mL).
- LLoQ: 50 IU/mL (1.70 Log IU/mL) with TAE and TE less than or equal to 1.00 Log IU/mL.
- Carryover: 0.3% (95% CI: 0.1% to 1.1%) resulting in detectable concentration >= LoD. 1.2% (95% CI: 0.6% to 2.4%) resulting in EBV detection.
- Negative Percent Agreement (NPA) with Comparator (confirmed EBV DNA negative clinical specimens): 100.0% (44/44) with a 95% confidence interval of (92.0%, 100.0%).
- Deming Regression Analysis: slope of 1.01, intercept of 0.05, correlation coefficient of 0.974.
- Mean Bias (Alinity m EBV minus comparator): 0.09 Log IU/mL with a 95% CI of (0.06, 0.12).
Predicate Device(s)
cobas® EBV, DEN200015
Reference Device(s)
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information
Not Found
§ 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.
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July 15, 2022
Image /page/0/Picture/1 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: the Department of Health and Human Services seal on the left and the FDA acronym and name on the right. The seal is a stylized depiction of an eagle and a caduceus, while the FDA acronym and name are written in blue, with the word "Administration" appearing below the acronym and name.
Abbott Molecular, Inc. Gina Sammarco Senior Specialist Regulatory Affairs 1300 E. Touhy Ave Des Plaines, Illinois 60018
Re: K212778
Trade/Device Name: Alinity m EBV AMP Kit (List No. 09N43-095). Alinity m EBV CTRL Kit (List No. 09N43-085), Alinity m EBV CAL Kit (List No. 09N43-075) 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: August 31, 2021 Received: September 1, 2021
Dear Gina Sammarco:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's
1
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 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.
For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely,
Maria Garcia, Ph.D. Assistant Director Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health 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) K212778
Device Name Alinity m EBV (09N43)
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 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 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) |
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Section 5: 510(k) Summary
Table of Contents
| 5.0 | 510(k) Summary | Page 2 |
|---|---|---|
| 5.1 | Submitter | 2 |
| 5.2 | Device Information | 3 |
| 5.3 | Predicate Device | 3 |
| 5.4 | Device Description | 4 |
| 5.5 | Intended Use | 6 |
| 5.6 | Similarities and Differences to Predicate Devices | 6 |
| 5.7 | Performance Data | 10 |
| 5.7.1 Specific Performance Characteristics | 10 | |
| 5.7.2 Clinical Reproducibility | 26 | |
| 5.7.3 Clinical Performance | 29 | |
| 5.8 | Conclusions Drawn from the Studies | 35 |
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5.0 510(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).
5.1 Submitter
| Applicants Name and Address: | Abbott Molecular Inc.
1300 E. Touhy Ave
Des Plaines, IL 60018 |
|------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Contact Person: | Gina Sammarco
Manager Regulatory Affairs
Abbott Molecular, Inc.
1300 E. Touhy Avenue
Des Plaines, IL 60018
Phone: 224-361-7627
Fax: 224-361-7646 |
| Date Prepared: | August 31, 2021 |
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| Trade Name | Regulation Name | Product
Code | Regulation
Number | Class |
|---------------|---------------------------------------------------------------------------|-----------------|----------------------|-------|
| Alinity m EBV | Quantitative viral nucleic acid test
for transplant patient management | QLX | 866.3183 | II |
Common Name: Nucleic acid test for quantitation of EBV DNA
5.3 Predicate Device
| Device Name | Predicate Device | De Novo | Cleared |
|---|---|---|---|
| Alinity m EBV | cobas® EBV | DEN200015 | July 30, 2020 |
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5.4 Device Description
Alinity m EBV is an in vitro polymerase chain reaction (PCR) assay for the quantitation of EBV DNA in human plasma.
The steps of the Alinity m EBV assay 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 multi-well amplification trays (AMP Trays) containing lyophilized, unit-dose PCR amplification/detection reagents and multi-well activation trays (ACT Trays) containing liquid, unit-dose activation reagents (MgCl2, TMAC, KCl, and ProClin). 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 a negative control, a low positive control, and a high positive control, each supplied as liquid in single-use tubes. The intended storage condition for the Alinity m EBV CTRL Kit is —15°C to —25°C.
- . Alinity m EBV CAL Kit (List No. 09N43-075) consisting of two levels of calibrators (CAL A and CAL B), each supplied as liquid in single-use tubes. The intended storage condition for the Alinity m EBV CAL Kit is -15°C to -25°C.
EBV DNA from specimens 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 the liquid unit-dose Alinity m EBV activation reagent and lyophilized unit-dose Alinity m EBV amplification/detection reagents and transferred into a reaction
7
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.
An EBV calibration curve is required for the quantitation of EBV targets. Two levels of calibrators are processed through sample preparation and real-time 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 remain satisfactory. During each control event, a negative control, a low-positive control, and a high-positive control are processed through sample preparation and real-time PCR procedures that are identical to those used for specimens.
At the beginning of the Alinity m EBV sample preparation process, a lyophilized unit -dose of Internal Control on the AMP Tray is rehydrated by the Alinity m System and delivered into each sample preparation reaction. The Internal Control 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 assay validity.
The Alinity m EBV amplification and detection reagents include primers and probes that amplify and detect dual targets in the EBV genome. Amplification and detection of the two EBV targets ensures sensitive detection of the viral genome even at low levels.
The Alinity m EBV assay also utilizes the following accessories:
- . Alinity m EBV Application Specification File, List No. 09N43-05A
- . 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
8
- . 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
- . 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
ર્સ્ટ Intended Use
Alinity m EBV AMP Kit
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.
5.6 Similarities and Differences to Predicate Devices
The primary functional components of the Alinity m EBV assay are substantially equivalent to other legally marketed nucleic acid amplification tests (NAAT) intended for the quantitative detection of EBV DNA.
9
The Alinity m EBV assay has the same general intended use as the predicate device. Although there are some technological differences between the Alinity m EBV assay and the predicate device, these differences do 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 EBV assay and the predicate device are shown in Table 1.
10
| Table 1. Similarities and Differences Between Alinity m EBV and Predicate Device | ||
|---|---|---|
| Device & | ||
| Predicate | ||
| Device(s): | Alinity m EBV | |
| K212778 | cobas EBV test | |
| DEN200015 | ||
| General Device | ||
| Characteristic | ||
| Similarities | ||
| Assay Type | Quantitative | Quantitative |
| Specimen Types | EDTA Plasma | EDTA Plasma |
| Sample | ||
| Preparation | ||
| Procedure | Automated liquid handling and robotic manipulation platform. | Automated liquid handling and robotic manipulation platform. |
| Amplification | ||
| Technology | Real-time polymerase chain reaction (PCR) | PCR |
| Assay Controls | Negative Control Low Positive Control High Positive Control Internal Control | Negative Control Low Positive Control High Positive Control DNA Quantitation Standard (DNA-QS) |
| General Device | ||
| Characteristic | ||
| Differences | ||
| 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. | cobas EBV is an in vitro nucleic acid amplification test for the | |
| quantitation of Epstein-Barr virus (EBV) DNA in human EDTA | ||
| plasma on the cobas 6800/8800 Systems. | ||
| 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. | cobas 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 response to treatment. | ||
| The results from cobas EBV are intended to be read and analyzed | ||
| by a qualified licensed healthcare professional in conjunction with | ||
| clinical signs and symptoms and relevant laboratory findings. | ||
| Negative test results do not preclude EBV infection or EBV | ||
| disease. Test results must not be the sole basis for patient | ||
| management decisions. cobas EBV is not intended for use as a | ||
| screening test for donors of blood or blood products or human | ||
| cells, tissues, and cellular and tissue-based products (HCT/Ps). | ||
| Assay Targets | 2 highly conserved regions of the EBV genome | EBV EBNA-1 gene |
| EBV BMRF gene |
CONFIDENTIAL
11
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5.7 Performance Data
The following performance data were provided in support of the safety, effectiveness and substantial equivalence determination of the device.
5.7.1 Specific Performance Characteristics
5.7.1.1 Traceability to the WHO Standard
Primary calibrators and assay product calibrators with known concentrations were used throughout product development and product manufacturing to establish traceability to the 1st World Health Organization (WHO) International Standard for Epstein-Barr virus for Nucleic Acid Amplification Techniques (NIBSC code: 09/260). The concentrations tested for the WHO standard were 3.00 Log IU/mL and 5.00 Log IU/mL. The target concentrations tested for the primary calibrators were 3.00 Log IU/mL and 7.00 Log IU/mL. The Alinity m EBV product calibrators and controls were also tested along with the primary calibrators and the WHO standard. All of the tested material had observed EBV concentrations similar to the target concentrations, and were linear across the assay's quantitation range, as presented in Figure 1.
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Image /page/13/Figure/1 description: This image is a scatter plot showing the relationship between the target concentration and the Alinity m EBV observed mean. The x-axis represents the target concentration in Log IU/mL, and the y-axis represents the Alinity m EBV observed mean in Log IU/mL. The plot includes data points for WHO, Primary Calibrator, Product Calibrator, and Control, and a dashed line representing the equation y = 1.00x - 0.02 with an R-squared value of 0.999.
Limit of Detection 5.7.1.2
The limit of detection (LoD) was determined for EBV type 1 by testing dilutions of the 1 st 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 4 lots of amplification reagents across multiple days. The results, representative of the analytical sensitivity performance of Alinity m EBV, are summarized in Table 2 (Lot 1), Table 3, (Lot 2), Table 4 (Lot 3), and Table 5 (Lot 4) .
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Probit analysis of the data using the least sensitive lot (Lot 3) determined that the concentration of EBV DNA in plasma detected with 95% probability (LoD by Probit) was 19.56 IU/mL with a 95% confidence interval (CI) of (13.09 IU/mL, 39.39 IU/mL) (Table 4).
| Table 2. Alinity m EBV Limit of Detection (LoD) - Lot 1 | |||
|---|---|---|---|
| EBV DNA | |||
| (IU/mL) | No. of Valid | ||
| Replicates | No. of Detected | ||
| Replicates | Detection | ||
| Rate (%) | |||
| 100.00 | 24 | 24 | 100.0 |
| 50.00 | 24 | 24 | 100.0 |
| 20.00 | 24 | 24 | 100.0 |
| 15.00 | 24 | 24 | 100.0 |
| 12.50 | 24 | 23 | 95.8 |
| 10.00 | 24 | 22 | 91.7 |
| 7.50 | 24 | 22 | 91.7 |
| 5.00 | 24 | 14 | 58.3 |
| 2.50 | 24 | 6 | 25.0 |
| 1.25 | 24 | 9 | 37.5 |
| Table 3. Alinity m EBV Limit of Detection (LoD) - Lot 2 | |||
|---|---|---|---|
| EBV DNA | |||
| (IU/mL) | No. of Valid | ||
| Replicates | No. of Detected | ||
| Replicates | Detection | ||
| Rate (%) | |||
| 100.00 | 24 | 24 | 100.0 |
| 50.00 | 24 | 24 | 100.0 |
| 20.00 | 24 | 23 | તે જેવી જેવી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં લોકોનો મુખ્ય વ્યવસાય ખેતી, ખેતમજૂરી તેમ જ પશુપાલન છે. આ ગામમાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી તેમ જ દૂધની ડેરી જેવી સવલ |
| 15.00 | 24 | 24 | 100.0 |
| 12.50 | 24 | 24 | 100.0 |
| 10.00 | 24 | 23 | તે જેવી જેવી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં લોકોનો મુખ્ય વ્યવસાય ખેતી, ખેતમજૂરી તેમ જ પશુપાલન છે. આ ગામમાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી તેમ જ દૂધની ડેરી જેવી સવલ |
| 7.50 | 24 | 21 | 87.5 |
| 5.00 | 24 | 17 | 70.8 |
| 2.50 | 24 | 13 | 54.2 |
| 1.25 | 24 | 9 | 37.5 |
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| Table 4. Alinity m EBV Limit of Detection (LoD) - Lot 3 (Least Sensitive Lot) | |||
|---|---|---|---|
| EBV DNA | |||
| (IU/mL) | No. of Valid | ||
| Replicates | No. of Detected | ||
| Replicates | Detection | ||
| Rate (%) | |||
| 100.00 | 24 | 24 | 100.0 |
| 50.00 | 24 | 24 | 100.0 |
| 20.00 | 24 | 24 | 100.0 |
| 15.00 | 23 | 22 | 95.7 |
| 12.50 | 23 | 20 | 87.0 |
| 10.00 | 24 | 21 | 87.5 |
| 7.50 | 24 | 19 | 79.2 |
| 5.00 | 24 | 16 | 66.7 |
| 2.50 | 24 | 9 | 37.5 |
| 1.25 | 24 | 7 | 29.2 |
| Table 5. Alinity m EBV Limit of Detection (LoD) - Lot 4 | |||
|---|---|---|---|
| EBV DNA | |||
| (IU/mL) | No. of Valid | ||
| Replicates | No. of Detected | ||
| Replicates | Detection | ||
| Rate (%) | |||
| 100.00 | 24 | 24 | 100.0 |
| 50.00 | 24 | 24 | 100.0 |
| 20.00 | 24 | 24 | 100.0 |
| 15.00 | 24 | 22 | 91.7 |
| 12.50 | 24 | 23 | 95.8 |
| 10.00 | 24 | 22 | 91.7 |
| 7.50 | 24 | 20 | 83.3 |
| 5.00 | 24 | 19 | 79.2 |
| 2.50 | 24 | 13 | 54.2 |
| 1.25 | 23 | 5 | 21.7 |
The claimed LoD of Alinity m EBV is 20 IU/mL (1.30 Log IU/mL) in plasma.
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5.7.1.3 Limit of Detection for EBV Type 2
Cultured virus for EBV type 2 was diluted to 3 different concentrations in EBV negative human plasma. Testing was performed using one lot of amplification reagents across multiple days. The results, representative of the analytical sensitivity performance of Alinity m EBV for EBV type 2, are summarized in Table 6.
Alinity m EBV detected 95% or greater of EBV samples at and above 15 IU/mL (1.18 Log IU/mL) in plasma. These results demonstrate the ability of Alinity m EBV to detect EBV type 2 at the claimed LoD.
| Table 6. Alinity m EBV Type 2 Limit of Detection (LoD) | |||
|---|---|---|---|
| EBV DNA | |||
| (IU/mL) | No. of Valid | ||
| Replicates | No. of Detected | ||
| Replicates | Detection | ||
| Rate (%) | |||
| 50.00 | 24 | 24 | 100.0 |
| 20.00 | 23 | 22 | 95.7 |
| 15.00 | 24 | 23 | 95.8 |
5.7.1.4 Linear Range
The quantitation range of Alinity m EBV is from the lower limit of quantitation (LLQ) of 50 IU/mL (1.70 Log IU/mL) to the upper limit of quantitation (ULoQ) of 200,000,000 IU/mL (8.30 Log IU/mL).
Linearity of Alinity m EBV was assessed by testing a dilution series of EBV type 1 in negative human plasma, consisting of 16 panel levels targeted in the range of 10 IU/mL to 400,000,000 IU/mL (1.00 Log IU/mL to 8.60 Log IU/mL). Panel levels with concentrations from 10 IU/mL to 1,500 IU/mL (1.00 Log IU/mL to 3.18 Log IU/mL) were prepared using clinical specimen, while panel levels with concentrations from 15 IU/mL to 400,000,000 IU/mL (1.18 Log IU/mL to 8.60 Log IU/mL) were prepared using synthetic DNA. Panel quantitation values were traceable to the 1st World Health Organization (WHO) International Standard for Epstein-Barr Virus for Nucleic Acid Amplification Techniques (NIBSC code: 09/260).
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Alinity m EBV was linear across the quantitation range from 50 IU/mL to 200.000.000 IU/mL (1.70 Log IU/mL to 8.30 Log IU/mL). Representative results for Alinity m EBV linearity performance are shown in Figure 2.
Image /page/17/Figure/1 description: The figure is a scatter plot that shows the correlation between Alinity m EBV (Log IU/mL) and Expected Concentration (Log IU/mL). The x-axis represents the Expected Concentration (Log IU/mL), and the y-axis represents the Alinity m EBV (Log IU/mL). The data points are clustered around a straight line, indicating a strong positive correlation. The correlation coefficient, r, is 0.999, which further confirms the strong positive correlation between the two variables.
Figure 2. Linearity a
3 The markers in the plot represent the mean Alinity m EBV concentration (in Log IU/mL) for each panel level.
Linearity of EBV Type 2 5.7.1.5
Linearity of Alinity m EBV for EBV type 2 was confirmed by testing a dilution series in negative human plasma, consisting of 16 panel levels targeted in the range of 10 IU/mL to 400,000,000 IU/mL (1.00 Log IU/mL to 8.60 Log IU/mL). Panel levels with concentrations from 10 IU/mL to 1,500 IU/mL (1.00 Log IU/mL to 3.18 Log IU/mL) were prepared using a cultured virus, while panel levels with concentrations from
18
15 IU/mL to 400,000,000 IU/mL (1.18 Log IU/mL to 8.60 Log IU/mL) were prepared using synthetic DNA. Panel quantitation values were traceable to the 1st World Health Organization (WHO) International Standard for Epstein-Barr Virus for Nucleic Acid Amplification Techniques (NIBSC code: 09/260).
Alinity m EBV was linear across the quantitation range from 50 IU/mL to 200,000,000 IU/mL (1.70 Log IU/mL to 8.30 Log IU/mL) for EBV type 2.
Representative results for Alinity m EBV linearity performance for type 2 and for type 1 (Section 5.7.1.4) are shown in Figure 3.
Image /page/18/Figure/3 description: The image shows the title of a figure. The title is "Figure 3. Linearity for EBV Type 1 and Type 2 ª". The title is written in a clear, sans-serif font and is centered on the page. The figure number is 3.
Image /page/18/Figure/4 description: The image is a scatter plot comparing expected concentration to Alinity m EBV. The x-axis is labeled 'Expected Concentration (Log IU/mL)' and ranges from 0 to 9. The y-axis is labeled 'Alinity m EBV (Log IU/mL)' and ranges from 0 to 9. There are two lines plotted on the graph, one for Type 1 and one for Type 2, both with r=0.999.
a The markers in the plot represent the mean Alinity m EBV concentration (in Log IU/mL) for each panel level.
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Alinity m EBV was demonstrated to be linear across the quantitation range for EBV type 1 and type 2 from 50 IU/mL to 200.000.000 IU/mL (1.70 Log IU/mL to 8.30 Log IU/mL).
5.7.1.6 Precision
Precision of Alinity m EBV was determined by analyzing a 9-member plasma panel. Panel members with concentrations targeted to 1.30 Log IU/mL and 2.00 Log IU/mL (20 IU/mL and 100 IU/mL) were prepared with positive clinical sample, panel members targeted in the range of 2.70 Log IU/mL to 5.00 Log IU/mL (500 IU/mL to 100,000 IU/mL) were prepared using cultured virus, and panel members with targeted concentrations greater than 5.00 Log IU/mL were prepared using synthetic DNA. Each panel member was tested in 4 replicates, twice each day for 12 days, on 3 Alinity m Systems operated by 3 operators (1 operator per instrument), using 3 AMP kit lots (1 lot per instrument), for a total of 288 replicates per panel member.
The representative precision study results in Table 8 demonstrated that Alinity m EBV within-laboratory standard deviation (SD) was less than or equal to 0.25 Log IU/mL for EBV DNA panels targeted in the range of 2.70 Log IU/mL to 8.30 Log IU/mL (500 IU/mL to 200.000.000 IU/mL), and less than or equal to 0.50 Log IU/mL for EBV DNA panels targeted in the range of 1.30 Log IU/mL to less than 2.70 Log IU/mL (20 IU/mL to less than 500 IU/mL).
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| Table 7. Precision | |||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Mean | |||||||||||||||
| Concentration | |||||||||||||||
| (Log IU/mL) | Within-Run | ||||||||||||||
| Component | Between-Run | ||||||||||||||
| Component | Between-Day | ||||||||||||||
| Component | Within- | ||||||||||||||
| Laboratoryc | Between- | ||||||||||||||
| Instrument | |||||||||||||||
| Componentd | Totale | ||||||||||||||
| Panel | Na | SDb | % CV | SD | % CV | SD | % CV | SD | % CV | SD | % CV | SD | % CV | ||
| 9 | 287 | 8.22 | 0.04 | 0.5 | 0.04 | 0.5 | 0.00 | 0.0 | 0.06 | 0.7 | 0.05 | 0.6 | 0.08 | 0.9 | |
| 8 | 287 | 8.04 | 0.05 | 0.6 | 0.04 | 0.4 | 0.00 | 0.0 | 0.06 | 0.7 | 0.06 | 0.8 | 0.09 | 1.1 | |
| 7 | 288 | 7.05 | 0.05 | 0.7 | 0.02 | 0.3 | 0.00 | 0.0 | 0.05 | 0.7 | 0.02 | 0.4 | 0.06 | 0.8 | |
| 6 | 284 | 6.09 | 0.05 | 0.8 | 0.03 | 0.4 | 0.00 | 0.0 | 0.06 | 0.9 | 0.04 | 0.7 | 0.07 | 1.1 | |
| 5 | 287 | 5.05 | 0.05 | 0.9 | 0.03 | 0.6 | 0.01 | 0.1 | 0.05 | 1.1 | 0.05 | 0.9 | 0.07 | 1.4 | |
| 4 | 287 | 4.06 | 0.04 | 1.1 | 0.03 | 0.7 | 0.02 | 0.4 | 0.05 | 1.3 | 0.06 | 1.4 | 0.08 | 1.9 | |
| 3 | 288 | 2.77 | 0.07 | 2.7 | 0.07 | 2.7 | 0.00 | 0.0 | 0.11 | 3.8 | 0.08 | 3.0 | 0.13 | 4.9 | |
| 2 | 286 | 2.20 | 0.13 | 5.7 | 0.13 | 6.1 | 0.00 | 0.0 | 0.18 | 8.3 | 0.07 | 3.1 | 0.20 | 8.9 | |
| 1 | 283 | 1.43 | 0.25 | 17.2 | 0.07 | 5.2 | 0.01 | 0.6 | 0.26 | 18.0 | 0.05 | 3.7 | 0.26 | 18.4 |
ª Number of valid replicates with detectable viral load
b Standard deviations (SD) are in Log IU/mL.
· Within-Laboratory includes Within-Run, Between-Run, and Between-Day components.
d Alinity m System, AMP Kit lot, and Operator are confounding effect is represented by Instrument.
e Total includes Within-Run, Between-Run, Between-Day, and Between-Instrument Components.
Alinity m EBV 510(k) Section 5 May 2022 K212778 510k summary FINAL.docx CONFIDENTIAL
21
| Table 8. Precision | |||||||
|---|---|---|---|---|---|---|---|
| CV(%)c | |||||||
| Panel | Na | Mean | |||||
| Concentrationb | |||||||
| (IU/mL) | Within-Run | ||||||
| Component | Between-Run | ||||||
| Component | Between-Day | ||||||
| Component | Between- | ||||||
| Instrument | |||||||
| Componentd | Totale | ||||||
| 9 | 287 | 169,046,883 | 10.0 | 9.5 | 0.0 | 10.9 | 17.7 |
| 8 | 287 | 111,475,841 | 10.7 | 8.2 | 0.0 | 14.9 | 20.2 |
| 7 | 288 | 11,275,291 | 10.8 | 4.3 | 0.0 | 5.7 | 13.0 |
| 6 | 284 | 1,231,685 | 11.4 | 6.1 | 0.0 | 9.4 | 16.0 |
| 5 | 287 | 112,523 | 10.7 | 6.6 | 1.2 | 10.9 | 16.8 |
| 4 | 287 | 11,596 | 10.0 | 6.3 | 3.5 | 13.3 | 18.2 |
| 3 | 288 | 620 | 17.4 | 17.2 | 0.0 | 19.5 | 31.8 |
| 2 | 286 | 176 | 29.5 | 31.5 | 0.0 | 16.0 | 47.5 |
| 1 | 283 | 32 | 61.3 | 17.2 | 2.0 | 12.3 | 66.2 |
Number of valid replicates with detectable viral load
b Titer data are considered to be log-normally distributed and the mean values for titer data are calculated as exp(mean * In(10) + [SD * In(10) (22).
& Titer data are considered to be log-normally distributed and %CV (%) = sqrt(10"[SD^2 * In(10)] = 1) * 100.
d Alinity m System, AMP Kit lot, and Operator are confounding effect is represented by Instrument.
e Total includes Within-Run, Between-Run, Between-Day, and Between-Instrument Components.
Alinity m EBV 510(k) Section 5 May 2022 K212778 510k summary FINAL.docx CONFIDENTIAL
22
5.7.1.7 Lower Limit of Quantitation
The lower limit of quantitation (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 by 2 methods:
- Total Analytical Error (TAE) = |bias| + 2 × SD, and .
- . Total Error (TE) = SORT(2) × 2 × SD.
The results of the calculations are shown in Table 9.
Panel members were 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 plasma.
The results of these analyses support a claimed LLoQ of 50.00 IU/mL (1.70 Log IU/mL) for Alinity m EBV, with an acceptable level of accuracy and precision, ie, TAE and TE less than or equal to 1.00 Log IU/mL.
| Table 9. Total Error | |||||
|---|---|---|---|---|---|
| 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) | |||||
| 1.30 | 1.34 | 0.04 | 0.27 | 0.59 | 0.78 |
| 1.70 | 1.77 | 0.07 | 0.22 | 0.51 | 0.62 |
| 2.00 | 2.12 | 0.12 | 0.14 | 0.40 | 0.40 |
ª Mean concentration - target concentration
Analytical Specificity – Potential Cross-Reactants 5.7.1.8
The analytical specificity of Alinity m EBV was evaluated with a panel of microorganisms (see Table 10) in EBV negative plasma, positive plasma targeted to 60 IU/mL EBV DNA, and positive plasma targeted to 10,000 IU/mL EBV DNA. Microorganisms were tested at a final concentration of 105 Units/mL for viruses and fungi or 106 Units/mL for bacteria. No cross-reactivity or interference in the performance of the Alinity m EBV assay was observed in the presence of the tested microorganisms.
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| Table 10. Potential Cross-Reactants | ||
|---|---|---|
| Viruses | Bacteria | Fungi |
| Adenovirus 2 | Actinomyces israelii | Aspergillus niger |
| BK polyomavirus | Clostridium perfringens | Candida albicans (CA) |
| Cytomegalovirus (CMV) | Enterococcus faecalis | Cryptococcus neoformans |
| Enterovirus Type 71 | Escherichia coli | |
| Hepatitis A Virus (HAV) | Klebsiella pneumoniae | |
| Hepatitis B Virus (HBV) | Listeria monocytogenes | |
| Hepatitis C Virus (HCV) | Morganella morganii | |
| Herpesvirus 6A | Mycobacterium smegmatis | |
| Herpesvirus 6B | Mycoplasma pneumoniae | |
| Herpesvirus 7 | Pseudomonas aeruginosa | |
| Herpesvirus 8 | ||
| (Kaposi's sarcoma associated virus) | Salmonella enterica | |
| Human immunodeficiency virus 1 (HIV-1) | Staphylococcus aureus (SA) | |
| Human immunodeficiency virus 2 (HIV-2) | Staphylococcus epidermidis | |
| Human papilloma virus 16 | ||
| (HPV-16) | Streptococcus pneumoniae | |
| Human papilloma virus 18 | ||
| (HPV-18) | ||
| Herpes Simplex Virus-1 (HSV-1) | ||
| Human T-lymphotropic virus type 1 | ||
| (HTLV-1) | ||
| Mumps orthorubulavirus | ||
| Parvo virus B19 | ||
| Simian Virus 40 | ||
| Vaccinia virus (VACV) | ||
| Varicella-Zoster virus (VZV) |
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5.7.1.9 Analytical Specificity - Potentially Interfering Substances
The effects of endogenous substances and the presence of high levels of therapeutic drugs commonly prescribed in transplant patients were evaluated. Potential interference on Alinity m EBV performance in plasma was assessed by testing 8 negative samples, 8 positive samples targeted to 60 IU/mL and 8 positive samples targeted to 10,000 IU/mL EBV DNA.
No interference was observed in the presence of albumin (60 g/L), hemoglobin (10 g/L), triglycerides (16.94 mmol/L), conjugated bilirubin (475 µmol/L), unconjugated bilirubin (684 umol/L), or human genomic DNA (2 µg/mL) that were introduced in the sample.
No interference was observed in the presence of drug compounds tested in pools or individually that are listed in Table 11, at a concentration of 3 times the reported Cmax or higher.
| Table 11. Drug Compounds | |
|---|---|
| Pools Tested | Drug Compounds |
| 1 | Mycophenolic acid |
| 2 | Amoxicillin |
| Clavulanate | |
| Foscarnet | |
| Piperacillin | |
| Tazobactam sodium | |
| Vancomycin | |
| 3 | Acyclovir |
| Amlodipine besylate | |
| Atenolol | |
| Azathioprine | |
| Cefotetan | |
| Cyclosporine | |
| Everolimus | |
| Famotidine | |
| Fluconazole | |
| Lisinopril | |
| Mycophenolate mofetil | |
| Prednisone | |
| Rabeprazole | |
| Sirolimus | |
| Sulfamethoxazole | |
| Tacrolimus | |
| Trimethoprim | |
| Valacyclovir | |
| Valsartan |
25
5.7.1.10 Carryover
The carryover rate for Alinity m EBV was determined by analyzing 648 valid replicates of EBV negative samples processed from alternating positions with 647 valid replicates of high concentrated EBV positive samples greater than or equal to 20,000,000 IU/mL, across a minimum of 27 runs. The carryover resulting in a detectable concentration greater than or equal to LoD was 0.3% (95% CI: 0.1% to 1.1%). The carryover resulting in EBV detection was 1.2% (95% CI: 0.6% to 2.4%).
5.7.1.11 Alinity m EBV Testing Using Dilution Procedure
The 1:2.5 dilution procedure was evaluated by comparing quantitation of neat samples and samples tested using the Alinity m EBV dilution procedure. Five plasma panel members with EBV levels targeted in the range of 150 IU/mL to 100,000,000 IU/mL (2.18 Log IU/mL to 8.00 Log IU/mL) were tested. Each panel member was tested, neat or using the dilution procedures, in multiple replicates. For the 5 panel members, the differences in mean, ie, diluted minus neat, ranged from -0.01 Log IU/mL to 0.23 Log IU/mL.
5.7.1.12 Precision of Alinity m EBV Using Dilution Procedure
Precision of Alinity m EBV, using the dilution procedure, was determined by analyzing 3 plasma panel members. Panel members 1 and 2 were prepared by spiking cultured virus in EBV negative sample, and panel member 3 was prepared by spiking synthetic DNA in EBV negative sample. Each panel member was tested in at least 3 replicates, twice each day for 12 days, on 3 Alinity m Systems with 3 Alinity m Specimen Dilution Kit I lots and 3 Alinity m EBV AMP Kit lots by 3 operators (1 Specimen Diluent lot, 1 AMP kit lot and 1 operator per instrument), for a total of at least 216 replicates.
The results, representative of the precision of Alinity m EBV using dilution procedures, are summarized in Table 12 and Table 13.
26
| | | Mean
Concentration | Within-Run
Component | | Between-Run
Component | | Between-Day
Component | | Within-Laboratoryc | | Between-Instrument
Componentd | | Totale | |
|-------|-----|-----------------------|-------------------------|------|--------------------------|------|--------------------------|------|--------------------|------|----------------------------------|------|--------|------|
| Panel | Na | (Log IU/mL) | SDb | % CV | SD | % CV | SD | % CV | SD | % CV | SD | % CV | SD | % CV |
| 1 | 274 | 3.50 | 0.07 | 2.1 | 0.06 | 1.6 | 0.06 | 1.7 | 0.11 | 3.2 | 0.04 | 1.2 | 0.12 | 3.4 |
| 2 | 273 | 4.89 | 0.05 | 1.1 | 0.09 | 1.9 | 0.00 | 0.0 | 0.11 | 2.2 | 0.06 | 1.1 | 0.12 | 2.5 |
| 3 | 274 | 7.69 | 0.06 | 0.8 | 0.09 | 1.2 | 0.00 | 0.0 | 0.11 | 1.4 | 0.04 | 0.5 | 0.12 | 1.5 |
ª Number of valid replicates with detectable viral load.
b Standard deviations (SD) are in Log IU/mL.
& Within-Laboratory includes Within-Run, Between-Run and Between-Day Components.
d Alinity m System, AMP Kit lot, Specimen Diluent lot, and Operator are confounding effect is represented by Instrument.
e Total includes Within-Run, Between-Run, Between-Day, and Between-Instrument Components.
| Table 13. Precision Using Dilution Procedure | |||||||
|---|---|---|---|---|---|---|---|
| Panel | Na | Mean | |||||
| Concentrationb | |||||||
| (IU/mL) | CV(%)c | ||||||
| Within-Run | |||||||
| Component | Between-Run | ||||||
| Component | Between-Day | ||||||
| Component | Between-Instrument | ||||||
| Componentd | Totale | ||||||
| 1 | 274 | 3,263 | 17.0 | 13.3 | 13.7 | 9.7 | 27.7 |
| 2 | 273 | 81,253 | 12.6 | 21.7 | 0.0 | 12.9 | 28.5 |
| 3 | 274 | 51,057,729 | 14.4 | 21.2 | 0.0 | 9.6 | 27.6 |
ª Number of valid replicates with detectable viral load
b Titer data are considered to be log-normally distributed and the mean values for titer data are calculated as exp(mean * In(10) + [SD * In(10) (22).
& Titer data are considered to be log-normally distributed and %CV (%)= sqrt(10°[SD^2 * In(10)]= 1) * 100.
d Alinity m System, AMP Kit lot, and Operator are confounding effect is represented by Instrument.
Page 24 of 35
e Total includes Within-Run, Between-Run, Between-Day, and Between-Instrument Components.
CONFIDENTIAL
27
5.7.1.13 Confirmation of the LLoQ Using Dilution Procedure
Confirmation testing for Alinity m EBV LLoQ using the dilution procedure was performed by testing 2 panel members at 50 IU/mL (LLoQ) and 60 IU/mL (near LLoQ) with a dilution factor of 1:2.5. The EBV concentrations in the panel members were targeted at 20 IU/mL and 24 IU/mL (1.30 Log IU/mL) after dilution in Specimen Diluent. Panel members were dilutions of cultured virus spiked into EBV negative plasma.
| Panel | Target
Concentration
Undiluted
(Log IU/mL) | Dilution
Factor | Target
Concentration
in Specimen
Diluent
(Log IU/mL) | Mean
Concentration a
(Log IU/mL) | Bias b
(Log IU/mL) | SD
(Log IU/mL) | TAE
(Log IU/mL) | TE
(Log IU/mL) |
|-------|-----------------------------------------------------|--------------------|------------------------------------------------------------------|----------------------------------------|-----------------------|-------------------|--------------------|-------------------|
| 1 | 1.70 | 2.5 | 1.30 | 1.71 | 0.01 | 0.17 | 0.35 | 0.48 |
| 2 | 1.78 | 2.5 | 1.38 | 1.74 | -0.04 | 0.23 | 0.50 | 0.65 |
ª Reported concentration for undiluted samples.
b Mean concentration - target concentration for undiluted samples
A minimum of 14 replicates per day of each panel level were tested using the dilution procedure in 3 runs across 3 days (one run per day). The study was performed using 1 Alinity m EBV AMP Kit lot, 1 Specimen Diluent lot, and 1 Alinity m System. Total error was estimated by TAE and TE, as shown in Table 14. The accuracy and precision at 20 IU/mL were confirmed for Alinity m EBV testing using the 1:2.5 dilution procedure.
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5.7.2 Clinical Reproducibility
Reproducibility performance of Alinity m EBV was evaluated by testing a 9-member reproducibility panel, including 8 positive panel members and 1 negative panel member. The positive panel members were prepared using an EBV positive clinical specimen, cultured virus, or plasmid DNA diluted in human EDTA plasma. The concentration levels targeted for the reproducibility panels spanned the quantitation range of the assay. A total of 3 Alinity m EBV AMP Kit lots, 3 Alinity m EBV CAL Kit lots, 3 Alinity m EBV CTRL Kit lots and 3 Alinity m Sample Prep Kit 2 lots were used. Three clinical sites each tested 2 unique reagent lot combinations (consisting of 2 Alinity m EBV AMP Kit lots along with 1 lot of each Alinity m EBV CAL Kit, Alinity m EBV CTRL Kit and Alinity m Sample Prep Kit 2) on 5 non-consecutive days for each lot combination. Six replicates of each panel member were tested on each of 5 days to ensure a minimum of 5 valid replicates for analysis. The reproducibility results are summarized in Table 15 and Table 16 (for the positive panel members) and Table 17 (for the negative panel member).
29
| Table 15. Reproducibility for Positive Panel Members | |||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Panel | Na | Target | |||||||||||||
| Concentration | |||||||||||||||
| (Log IU/mL) | Mean | ||||||||||||||
| Concentration | |||||||||||||||
| (Log IU/mL) | Within-Run/Day | ||||||||||||||
| Component | Between-Run/Day | ||||||||||||||
| Component | Within- | ||||||||||||||
| Laboratoryc | Between-Lot | ||||||||||||||
| Component | Between-Site/ | ||||||||||||||
| Instrument | |||||||||||||||
| Component | Totald | ||||||||||||||
| SDb | % CV | SD | % CV | SD | % CV | SD | % CV | SD | % CV | SD | % CV | ||||
| 1 | 178 | 8.30 | 8.40 | 0.05 | 0.5 | 0.03 | 0.3 | 0.05 | 0.6 | 0.06 | 0.7 | 0.18 | 2.1 | 0.19 | 2.3 |
| 2 | 177 | 7.00 | 7.04 | 0.04 | 0.6 | 0.00 | 0.1 | 0.04 | 0.6 | 0.04 | 0.6 | 0.12 | 1.8 | 0.14 | 2.0 |
| 3 | 179 | 6.00 | 5.76 | 0.05 | 0.8 | 0.02 | 0.3 | 0.05 | 0.9 | 0.04 | 0.7 | 0.08 | 1.3 | 0.10 | 1.8 |
| 4 | 179 | 5.00 | 5.04 | 0.06 | 1.1 | 0.04 | 0.9 | 0.07 | 1.4 | 0.04 | 0.7 | 0.09 | 1.8 | 0.12 | 2.4 |
| 5 | 179 | 4.00 | 4.01 | 0.05 | 1.2 | 0.03 | 0.9 | 0.06 | 1.5 | 0.04 | 1.0 | 0.05 | 1.3 | 0.09 | 2.2 |
| 6 | 172 | 3.00 | 3.07 | 0.07 | 2.2 | 0.03 | 1.0 | 0.07 | 2.4 | 0.02 | 0.7 | 0.04 | 1.3 | 0.09 | 2.8 |
| 7 | 179 | 1.78 | 1.64 | 0.19 | 11.4 | 0.02 | 1.0 | 0.19 | 11.5 | 0.04 | 2.4 | 0.13 | 7.6 | 0.23 | 14.0 |
| 8 | 174 | 1.30 | 1.20 | 0.28 | 23.5 | 0.05 | 4.2 | 0.29 | 23.8 | 0.00 | 0.0 | 0.10 | 8.0 | 0.30 | 25.1 |
4 Number of valid replicates with detectable viral load
6 Standard deviations (SD) are in Log IU/mL.
? Within-Laboratory includes Within-Run/Day and Between-Run/Day components.
Total includes Within-Run/Day, Between-Run/Day, Between-Lot, and Between-Site/Instrument Components.
CONFIDENTIAL
30
| CV(%)c | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Panel | Na | Mean | |||||||
| Concentrationb | |||||||||
| (IU/mL) | Within- | ||||||||
| Run/Day | |||||||||
| Component | Between- | ||||||||
| Run/Day | |||||||||
| Component | Between-Lot | ||||||||
| Component | Between-Site/ | ||||||||
| Instrument | |||||||||
| Component | Total | ||||||||
| 1 | 178 | 279,192,220 | 10.6 | 6.4 | 13.0 | 42.8 | 47.1 | ||
| 2 | 177 | 11,407,341 | 9.5 | 1.1 | 9.3 | 29.3 | 32.5 | ||
| 3 | 179 | 586,126 | 11.3 | 3.9 | 9.4 | 18.0 | 23.7 | ||
| 4 | 179 | 112,772 | 13.1 | 10.1 | 8.2 | 21.4 | 28.6 | ||
| 5 | 179 | 10,572 | 11.6 | 8.0 | 9.7 | 11.7 | 20.8 | ||
| 6 | 172 | 1,187 | 15.3 | 6.9 | 4.9 | 9.3 | 19.9 | ||
| 7 | 179 | 51 | 45.4 | 3.9 | 9.0 | 29.4 | 56.8 | ||
| 8 | 174 | 20 | 72.1 | 11.5 | 0.0 | 22.3 | 78.6 |
CONFIDENTIAL
31
| Table 17. Reproducibility for Negative Panel Member | ||||
|---|---|---|---|---|
| Expected EBV DNA Concentration | No. of Replicates | Negative Rate (%) | 95% Confidence Interval | |
| Valid | Negative | |||
| Negative | 180 | 176 | 97.8 (176/180) | (94.4, 99.1) |
5.7.3 Clinical Performance
Alinity m EBV results were compared to those of an FDA-cleared EBV nucleic acid test in a representative study. A total of 558 EDTA plasma samples were tested (neat or diluted), including 542 clinical specimens from hematopoietic stem cell transplant (HSCT) and solid organ transplant (SOT) subjects, and 16 contrived samples prepared by spiking inactivated EBV virus into the individual clinical specimens. The Alinity m EBV assay testing was performed at 3 clinical testing sites with 3 Alinity m EBV reagent kit lots.
Of the 558 clinical samples, 550 produced valid results with Alinity m EBV and the comparator assay, including 388 samples detected by Alinity m EBV and 162 samples not detected by Alinity m EBV. Out of 550 valid samples, 168 were from HSCT subjects, 379 were from SOT subjects, and 3 were from dual-transplant (HSCT/SOT) subjects. The agreement between Alinity m EBV and comparator results is shown in Table 18 (HSCT samples), Table 19 (SOT samples), and Table 20 (HSCT and SOT samples combined).
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| Table 18. HSCT Samples – Agreement Between Alinity m EBV and Comparatora | ||||||||
|---|---|---|---|---|---|---|---|---|
| Comparator EBV (Log IU/mL) | ||||||||
| Alinity m EBV | ||||||||
| (Log IU/mL) | Target | |||||||
| Not | ||||||||
| Detected |