Immunoassay for the in vitro quantitative determination of c1-fetoprotein in human serum and in the management of patients with non-seminomatous germ cell tumors.

The electrochemiluminescence immunoassay "ECLIA" is intended for use on the cobas e immunoassay analyzers.

Device Description

Immunoassay for the in vitro quantitative determination of αι-fetoprotein in human serum and plasma to aid in the management of patients with non-seminomatous germ cell tumors.

The electrochemiluminescence immunoassay "ECLIA" is intended for use on cobas e immunoassay analyzers.

Elecsys AFP utilizes a sandwich test principle and has a total test duration of 18 minutes.

1st incubation: 10 uL of sample, a biotinylated monoclonal AFP specific antibody, and . a monoclonal AFP specific antibody labeled with a ruthenium complex® react to form a sandwich complex.
2nd incubation: After addition of streptavidin-coated microparticles, the complex . becomes bound to the solid phase via interaction of biotin and streptavidin.
The reaction mixture is aspirated into the measuring cell where the microparticles are . magnetically captured onto the surface of the electrode. Unbound substances are then removed with ProCell/ProCell M. Application of a voltage to the electrode then induces chemiluminescent emission which is measured by a photomultiplier.
Results are determined via a calibration curve which is instrument-specifically . generated by 2 point calibration and a master curve provided via the reagent barcode or e barcode.
a) Tris(2,2'-bipyridyl)ruthenium(II)-complex (Ru(bpy) )

The reagent rackpack (M, R1, R2) is labeled as AFP:

M: Streptavidin-coated microparticles (transparent cap), 1 bottle. 12 mL: Streptavidin-. coated microparticles 0.72 mg/mL; preservative.
R1: Anti-AFP-Ab~biotin (gray cap), 1 bottle, 17 mL: Biotinylated monoclonal anti-. AFP antibodies (mouse) 4.5 mg/L; phosphate buffer 100 mmol/L, pH 6.0; preservative.
R2: Anti-AFP-Ab~Ru(bpy) (black cap), 1 bottle, 17 mL: Monoclonal anti-AFP . antibodies (mouse) labeled with ruthenium complex 12.0 mg/L; phosphate buffer 100 mmol/L, pH 6.0; preservative.

AI/ML Overview

The provided text describes a 510(k) premarket notification for the Elecsys AFP immunoassay. While it details numerous non-clinical tests conducted to prove the device's performance, it does not describe a study involving a "human-in-the-loop" or "algorithm-only" performance study in the context of an AI/ML device. The device described is an "immunological test system" and its performance is evaluated based on quantitative analytical metrics rather than the type of diagnostic performance (e.g., sensitivity, specificity, accuracy) typically associated with AI/ML devices interpreting medical images or complex data for diagnostic purposes.

Therefore, many of the requested items (e.g., number of experts, adjudication methods, MRMC studies, effect size of human reader improvement with AI) are not applicable or cannot be extracted from the provided text for this specific device.

However, I can extract information related to the acceptance criteria and performance of the described immunoassay device based on the non-clinical tests performed.

Here's a breakdown of the available information based on your request, focusing on what can be extracted and noting what cannot (due to the nature of the device and the provided document):

Device: Elecsys AFP Immunoassay (K220176)

Purpose of the study (as described in the document): To demonstrate substantial equivalence of the updated Elecsys AFP assay to a predicate device (K981282) by improving biotin tolerance and reducing streptavidin interference. The studies are non-clinical evaluations of the analytical performance of the immunoassay.

1. Table of Acceptance Criteria and Reported Device Performance

The document states that "All predefined acceptance criteria was met" for the various tests. While the specific numerical acceptance criteria (e.g., acceptable range for precision, linearity, interference limits) are not fully detailed, the reported performance is often given or implied by the "met" statement and the claimed range/limits.

Test Category	Specific Test	Acceptance Criteria (Implied/Stated)	Reported Device Performance/Conclusion
Precision	Repeatability (within-run)	Predefined criteria (CLSI EP05-A3)	All predefined acceptance criteria was met.
	Intermediate Precision (within-laboratory)	Predefined criteria (CLSI EP05-A3)	All predefined acceptance criteria was met.
Reproducibility	Lot-to-lot Reproducibility	Predefined criteria	All predefined acceptance criteria was met.
Analytical Sensitivity	Limit of Blank (LoB)	Determined according to CLSI EP17-A2	LoB claim in labeling: 0.75 IU/mL
	Limit of Detection (LoD)	Determined according to CLSI EP17-A2	LoD claim in labeling: 1.5 IU/mL
	Limit of Quantitation (LoQ)	Determined according to CLSI EP17-A2	LoQ claim in labeling: 1.5 IU/mL
Analytical Range	Linearity	Evaluated according to CLSI EP06-A (linear in defined range)	Linearity confirmed in range 0.58 IU/mL - 1129 IU/mL. Measuring range claimed: 1.50-1000 IU/mL.
	High-Dose Hook Effect (HDHE)	No hook effect observed up to a certain concentration	No hook effect seen up to 1,000,000 (concentration units not specified, likely referring to IU/mL).
Interference	Endogenous Substances (7 types)	Predefined criteria (acceptable interference limits)	All predefined acceptance criteria was met. Proposed labeling claims: Biotin ≤ 1200 ng/mL, etc.
	Exogenous/Pharmaceutical Compounds (27 types)	Predefined criteria (no interference observed)	All predefined acceptance criteria was met. No interference was observed.
Method Comparison	Candidate vs. Current Assay	Acceptable agreement (implied by regression and tau values)	Linear regression: y = -0.148 + 0.969x, r = 0.999. Passing/Bablok: y = 0.101 + 0.965x, tau = 0.985.
Sample Type	Anticoagulant Effect	Acceptable values compared to native serum	All predefined acceptance criteria was met, supporting use of serum, Li-Heparin, K2-EDTA, K3-EDTA plasma.
Stability	On-board Reagent Stability	Acceptable performance for claimed duration	Stable for up to 28 days (4 weeks) on analyzers.
	Lot Calibration Frequency	Acceptable performance for claimed duration	Calibration recommended every 4 weeks (1 month).

2. Sample Size Used for the Test Set and Data Provenance

Precision (Repeatability, Intermediate Precision): Not explicitly stated, but CLSI EP05-A3 typically involves multiple runs over several days with specific sample replicates.
Lot-to-lot Reproducibility: Used "three reagent lots."
Limit of Blank/Detection/Quantitation: Not explicitly stated, but CLSI EP17-A2 involves specific sample types and replicates.
Linearity: Evaluated on "one cobas e 601 analyzer" using samples spanning the linearity range, but the number of distinct samples or replicates is not specified beyond "0.58 IU/mL - 1129 IU/mL".
High-Dose Hook Effect: Assessed on "one cobas e 601 analyzer" in "two-fold determination" for "both samples."
Interference (Endogenous): "Seven endogenous substances were evaluated."
Interference (Exogenous): "17 commonly used and ten specially used pharmaceutical compounds" were evaluated.
Method Comparison: "A total of 181 serum sample concentrations were between 1.58 and 966 IU/mL."
Anticoagulant Effect: "native, single-donor samples drawn into serum, Li-Heparin, K2-EDTA, and K3-EDTA plasma primary tubes." (Number of samples or donors not specified).
Data Provenance: The document does not specify the country of origin for the samples/data or if they were retrospective or prospective. Given the nature of analytical validation studies for IVD devices, they are typically prospective studies performed in a controlled laboratory setting.

3. Number of Experts Used to Establish Ground Truth and Qualifications:

Not Applicable. This is an immunoassay, not an AI/ML device relying on expert interpretation for ground truth. The "ground truth" for these analytical studies is derived from established laboratory methods, reference materials, and defined analytical ranges.

4. Adjudication Method for the Test Set:

Not Applicable. No human adjudication process as typically seen in AI/ML diagnostic studies.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was done:

No. This is not an AI/ML device; therefore, an MRMC study comparing human readers with and without AI assistance is not relevant or was not performed.

6. If a Standalone (algorithm only without human-in-the-loop performance) was done:

Not Applicable (in the context of AI/ML). The device itself is an automated immunoassay. Its performance is inherently "standalone" in that it produces a quantitative result without human "interpretation" of a complex output, but this is a very different concept than an AI algorithm's standalone diagnostic performance. The document describes the analytical performance of the automated assay system.

7. The Type of Ground Truth Used:

The "ground truth" for these types of analytical studies is based on:
- Reference Methods/Materials: For linearity, analytical range, LoB/LoD/LoQ determinations, often using diluted/spiked samples with known concentrations.
- Comparative Analysis: The "method comparison" compares the new assay's results against the existing (predicate) assay.
- Defined Limits/Spiked Samples: For interference studies, known amounts of interferents are added to samples.
- Established CLSI Guidelines: The studies explicitly state adherence to CLSI (Clinical and Laboratory Standards Institute) guidelines (e.g., EP05-A3, EP17-A2, EP06-A) for methodology, which dictate how "truth" is assessed for analytical performance.

8. The Sample Size for the Training Set:

Not Applicable. This is not an AI/ML device that requires a "training set" in the computational sense. The device's performance is based on its chemical and technological design, and its validation involves analytical testing rather than machine learning on a dataset.

9. How the Ground Truth for the Training Set was Established:

Not Applicable. As above, no "training set" in the AI/ML context.

Summary

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September 15, 2022

Roche Diagnostics Jamie Ferguson Regulatory Affairs Principal 9115 Hague Road Indianapolis, Indiana 46250

Re: K220176

Trade/Device Name: Elecsys AFP Regulation Number: 21 CFR 866.6010 Regulation Name: Tumor-Associated Antigen Immunological Test System Regulatory Class: Class II Product Code: LOJ Dated: January 20, 2022 Received: January 21, 2022

Dear Jamie Ferguson:

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

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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,

Ying Mao, Ph.D. Branch Chief Division of Immunology and Hematology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K220176

Device Name Elecsys AFP

Indications for Use (Describe)

Immunoassay for the in vitro quantitative determination of c1-fetoprotein in human serum and in the management of patients with non-seminomatous germ cell tumors.

The electrochemiluminescence immunoassay "ECLIA" is intended for use on the cobas e immunoassay analyzers.

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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Elecsys AFP 510(k) Summary

This summary of 510(k) safety and effectiveness information is submitted in accordance with the requirements of 21 CFR 807.92.

In accordance with 21 CFR 807.87, Roche Diagnostics hereby submits official notification as required by Section 510(k) of the Federal Food, Drug and Cosmetics Act of our intention to market the device described in this Premarket Notification 510(k).

Submitter Name	Roche Diagnostics
Address	9115 Hague RoadP.O. Box 50416Indianapolis, IN 46250-0457
Contact	Contact: Jamie FergusonPhone: (317) 270-7402Email: Jamie.ferguson@roche.com
Date Prepared	September 15, 2022
Proprietary Name	Elecsys AFP
Common Name	Tumor-associated antigen immunological test system
Classification Name	Kit, Test, Alpha-Fetoprotein for Testicular Cancer
Product Codes,Regulation Numbers	LOJ, 21 CFR 866.6010
Predicate Devices	K981282, Elecsys AFP

The purpose of this Traditional 510(k) Premarket Notification is to obtain FDA review and clearance for the Elecsys AFP.

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1. DEVICE DESCRIPTION SUMMARY

Immunoassay for the in vitro quantitative determination of αι-fetoprotein in human serum and plasma to aid in the management of patients with non-seminomatous germ cell tumors.

The electrochemiluminescence immunoassay "ECLIA" is intended for use on cobas e immunoassay analyzers.

Elecsys AFP utilizes a sandwich test principle and has a total test duration of 18 minutes.

1st incubation: 10 uL of sample, a biotinylated monoclonal AFP specific antibody, and . a monoclonal AFP specific antibody labeled with a ruthenium complex® react to form a sandwich complex.
2nd incubation: After addition of streptavidin-coated microparticles, the complex . becomes bound to the solid phase via interaction of biotin and streptavidin.
The reaction mixture is aspirated into the measuring cell where the microparticles are . magnetically captured onto the surface of the electrode. Unbound substances are then removed with ProCell/ProCell M. Application of a voltage to the electrode then induces chemiluminescent emission which is measured by a photomultiplier.
Results are determined via a calibration curve which is instrument-specifically . generated by 2 point calibration and a master curve provided via the reagent barcode or e barcode.
a) Tris(2,2'-bipyridyl)ruthenium(II)-complex (Ru(bpy) )

The reagent rackpack (M, R1, R2) is labeled as AFP:

M: Streptavidin-coated microparticles (transparent cap), 1 bottle. 12 mL: Streptavidin-. coated microparticles 0.72 mg/mL; preservative.
R1: Anti-AFP-Ab~biotin (gray cap), 1 bottle, 17 mL: Biotinylated monoclonal anti-. AFP antibodies (mouse) 4.5 mg/L; phosphate buffer 100 mmol/L, pH 6.0; preservative.

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R2: Anti-AFP-Ab~Ru(bpy) (black cap), 1 bottle, 17 mL: Monoclonal anti-AFP . antibodies (mouse) labeled with ruthenium complex 12.0 mg/L; phosphate buffer 100 mmol/L, pH 6.0; preservative.

INTENDED USE/INDICATIONS FOR USE 2.

Immunoassay for the in vitro quantitative determination of a1-fetoprotein in human serum and plasma to aid in the management of patients with non-seminomatous germ cell tumors.

The electrochemiluminescence immunoassay "ECLIA" is intended for use on cobas e immunoassay analyzers.

INDICATIONS FOR USE COMPARISON 3.

Elecsys AFP (updated assay, Mat. No. 09015086160) is substantially equivalent to Elecsys AFP, cleared under K981282.

The intended use of Elecsys AFP was updated to remove analyzers that are no longer supported for use with Roche assays. The indications for use of Elecsys AFP did not change from the predicate device.

TECHNOLOGICAL COMPARISON 4.

Roche Diagnostics has updated the current Elecsys AFP assay in order to improve the biotin tolerance from ≤ 60 ng/mL to ≤ 1200 ng/mL and to reduce streptavidin interference. A technical solution was implemented by adding an anti-biotin antibody to one of the reagents, which allows depletion of biotin in patient samples by binding free biotin, and by adding a streptavidin interference reducing agent to enhance the streptavidin tolerance. No other technological characteristics were changed. The information submitted in this Premarket Notification supports a substantial equivalent decision.

NON-CLINICAL AND/OR CLINICAL TESTS SUMMARY & CONCLUSIONS 5.

Precision measurements were conducted for both 21 days and 5 days with the Elecsys AFP assay to evaluate repeatability (within-run precision) and intermediate precision (within-laboratory

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precision) according the CLSI guideline EP05-A3. All predefined acceptance criteria was met for the precision experiments.

Lot-to-lot reproducibility was performed for the Elecsys AFP assay using three reagent lots. All predefined acceptance criteria was met for the lot-to-lot reproducibility experiment.

The Limit of Blank (LoB) was determined according to CLSI EP17-A2. The LoB claim in the labeling will be set to 0.75 IU/mL.

The Limit of Detection (LoD) was determined according to CLSI EP17-A2. The LoD claim in the labeling will be set to 1.5 IU/mL.

The Limit of Quantitation (LoQ) was determined according to CLSI EP17-A2. The LoQ claim in the labeling will be set to 1.5 IU/mL.

Linearity was evaluated according to CLSI EP06-A with the Elecsys AFP assay on one cobas e 601 analyzer. Linearity was confirmed in the range of 0.58 IU/mL - 1129 IU/mL, and a measuring range of 1.50-1000 IU/mL will be claimed in the labeling.

The high-dose hook effect (HDHE) of the Elecsys AFP assay was assessed on one cobas e 601 analyzer in two-fold determination. No hook effect was seen up to 1,000,000 for both samples.

Seven endogenous substances were evaluated for potential interference with the Elecsys AFP assay on the cobas e 601 analyzer. All predefined acceptance criteria was met, and the proposed labeling claims for each endogenous substance can be found below:

Biotin ≤ 1200 ng/mL Lipemia (Intralipid) ≤ 1500 mg/dL Hemoglobin < 2200 mg/dL Bilirubin ≤ 65 mg/dL Rheumatoid Factor ≤ 1500 IU/mL Serum Albumin ≤ 7 g/dL IgG < 7 g/dL

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An exogenous interference study was conducted to evaluate 17 commonly used and ten specially used pharmaceutical compounds for potential interference with the Elecsys AFP assay on the cobas e 601 analyzer. The predefined acceptance criteria was met for all drugs tested, and no interference was observed.

A method comparison was performed with the current Elecsys AFP assay (material number 04481798190) and the candidate Elecsys AFP assay (material number 09015060160), using a total of 181 serum sample concentrations were between 1.58 and 966 IU/mL. The results can be found below:

Linear regression

v = -0.148 + 0.969x r = 0.999
Passing/Bablok

y = 0.101 + 0.965x

tau = 0.985

The effect on quantitation of analyte in the presence of anticoagulants with Elecsys AFP was determined by comparing values obtained from native, single-donor samples drawn into serum, Li-Heparin, K2-EDTA, and K3-EDTA plasma primary tubes. All predefined acceptance criteria was met, supporting the labeling claim that serum, Li-Heparin, K2-EDTA and K3-EDTA plasma primary tubes are acceptable sample types.

On-board reagent stability for the Elecsys AFP assay was tested on one cobas e 601 analyzer. Elecsys AFP reagent kits can be stored on-board the analyzers for up to 28 days (4 weeks). A new calibration of the kit kept on-board is recommended every 7 days.

Lot calibration frequency for the Elecsys AFP assay was tested on one cobas e 601 analyzer. Calibrations of an Elecsys AFP reagent lot is recommended every 4 weeks (1 month). During that time period, fresh reagent kits of the same lot can be used without calibration using the calibration curve of the day 0 reagent kit.

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The information provided in this 510(k) Premarket Notification supports the determination that the updated Elecsys AFP is substantially equivalent to the predicate device, Elecsys AFP (K981282).

Regulation Number and Section

§ 866.6010 Tumor-associated antigen immunological test system.

(a)
Identification. A tumor-associated antigen immunological test system is a device that consists of reagents used to qualitatively or quantitatively measure, by immunochemical techniques, tumor-associated antigens in serum, plasma, urine, or other body fluids. This device is intended as an aid in monitoring patients for disease progress or response to therapy or for the detection of recurrent or residual disease.(b)
Classification. Class II (special controls). Tumor markers must comply with the following special controls: (1) A guidance document entitled “Guidance Document for the Submission of Tumor Associated Antigen Premarket Notifications (510(k)s) to FDA,” and (2) voluntary assay performance standards issued by the National Committee on Clinical Laboratory Standards.