This is an indirect immunoenzyme antibody test for the semi-quantitative detection of antinuclear antibodies in human serum, with specific identification of autoantibodies to the SS-A/Ro antigen. This test system is to be used as an aid in the detection of antibodies associated with systemic rheumatic disease. The results from this assay can be used as an aid in the diagnosis of autoimmune diseases.

Device Description

This is an indirect immunoenzyme antibody test for the semi-quantitative detection of antinuclear antibody in human serum. The transfected HEP-2 cell line allows for the identification of anti-SS-A/Ro antibodies because of the unique staining pattern that these antibodies show on this cell line.

AI/ML Overview

The provided text describes a 510(k) submission for the HEp-2000® Colorzyme® ANA-Ro Test System, which is an indirect immunoenzyme antibody test. The document focuses on demonstrating substantial equivalence to a predicate device (RELISA® SS-A/SS-B Antibody Test System, K955603) rather than defining explicit acceptance criteria with numerical thresholds. However, we can infer some criteria from the performance descriptions.

1. Table of Acceptance Criteria and Reported Device Performance

Acceptance Criteria (Inferred)	Reported (Device) Performance
For Normal Samples: Minimal false positives and consistency with non-transfected HEp-2 cells in negative results.	Out of 500 normal samples, 36 (7.2%) were positive on both the device and non-transfected HEp-2 cells at 1:40 dilution. 34 out of 36 positive samples showed identical staining patterns. The 2 discrepant samples were confirmed to have anti-SS-A/Ro antibodies. Samples negative on ANA tests were also negative in ELISA.
For Samples with only Ro/SS-A Antibodies: High detection rate and distinctive staining pattern for Ro/SS-A.	Out of 46 samples with confirmed anti-SS-A/Ro antibodies: 100% were positive with the device (distinctive Ro/SS-A staining pattern). 78% were positive with non-transfected HEp-2 cells (speckled pattern).
For Samples with Other Autoantibodies (without Ro/SS-A): Consistency in staining patterns with non-transfected HEp-2 cells.	Out of 230 samples with various autoantibodies: 333 staining patterns were identical on both substrates. 29 samples showed distinctive "SS-A/Ro" on the device. 23 of these 29 showed speckled on non-transfected HEp-2. The 6 "discrepant" samples (positive on device, negative on non-transfected HEp-2) all had SS-A/Ro antibodies confirmed by ELISA and Western blot.
Titer Comparisons for Ro/SS-A Antibodies: Higher titer values compared to non-transfected HEp-2 cells.	Samples containing anti-Ro/SS-A antibodies show higher titer values on the device than on non-transfected HEp-2 cells due to overexpression.
Titer Comparisons for Other Autoantibodies: No significant titer differences compared to non-transfected HEp-2 cells.	Sera with other autoantibody specificities do not show significant titer differences between the device and non-transfected HEp-2 cells.
Reproducibility: Consistent qualitative results and minimal quantitative variation across lots and runs.	For 10 characterized samples run on 3 different lots on 3 occasions: No negative sample showed positive results. All titer values were within one two-fold dilution of the established mean titer value.
Confirmation of SS-A/Ro Antibodies (Clinical Utility): Distinctive SS-A/Ro pattern should correlate strongly with confirmed SS-A/Ro antibodies.	In a selected population of 349 ANA-positive samples, 239 showed the distinctive SS-A/Ro staining pattern with the device. 238 (99.6%) of these had positive ELISA tests for SS-A/Ro. 79 additional samples with strong speckled/homogeneous patterns also gave positive SS-A/Ro ELISA. Conclusion: Distinctive pattern is confirmatory, but its absence doesn't rule out SS-A/Ro.

2. Sample Sizes and Data Provenance

Normal Samples: 500 healthy blood donors (242 females, 258 males). Provenance: Not explicitly stated, likely clinical lab setting. Retrospective or Prospective: Not explicitly stated, but "were tested in parallel" suggests a batch analysis, which could be either.
Sera from Patients with Only Ro/SS-A Antibodies: 46 patients with SLE or Sjögren's Syndrome. Provenance: Not explicitly stated, likely clinical lab setting. Retrospective or Prospective: Not explicitly stated.
Sera from Patients with Autoantibodies other than SS-A/Ro: 230 patients with a variety of rheumatic and non-rheumatic diseases. Provenance: Not explicitly stated, likely clinical lab setting. Retrospective or Prospective: Not explicitly stated.
Reproducibility Titer: 10 samples (CDC controls and characterized in-house sera). Provenance: CDC controls and in-house laboratory. Retrospective or Prospective: Not explicitly stated, likely retrospective for established controls.
Confirmation SS-A/Ro Antibodies: 349 patients with known positive ANA tests (from a large rheumatology reference laboratory). Provenance: Clinical rheumatology reference laboratory. Retrospective or Prospective: Not explicitly stated, but "known positive ANA tests" suggests retrospective selection.

3. Number of Experts and Qualifications for Ground Truth

The document does not specify the "number of experts" used to establish ground truth in the context of interpretation of the device results (e.g., reading patterns). However, the ground truth for the presence of specific antibodies (SS-A/Ro) was established by ELISA testing and Western immunoblotting, which are established laboratory methodologies. The interpretation of these confirmatory tests would be performed by trained laboratory personnel, but no specific number or qualifications of "experts" are provided beyond implied laboratory competence.

4. Adjudication Method for the Test Set

No explicit adjudication method (e.g., 2+1, 3+1) for the test set results interpreted by the device is mentioned. The assessment often involved comparing the device's staining patterns to those on non-transfected HEp-2 cells, with discrepancies resolved by ELISA and Western immunoblotting (which serve as the ultimate ground truth).

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

No MRMC comparative effectiveness study involving human readers with and without AI assistance was reported. This device is an in vitro diagnostic test system that produces a staining pattern for visual interpretation, not an AI model providing automated interpretations or assistance. The comparison is between two different laboratory substrates.

6. Standalone (Algorithm Only) Performance Study

This is an in vitro diagnostic test kit that produces a visual result (staining pattern) to be interpreted by a human. It does not employ an AI algorithm in the standalone sense. The results are visual patterns on cells.

7. Type of Ground Truth Used

The ground truth used was primarily a combination of:

Confirmatory Laboratory Tests: ELISA testing and Western immunoblotting for the presence of specific autoantibodies (e.g., anti-SS-A/Ro).
Clinical Diagnoses: Patients were selected based on diagnoses like SLE or Sjögren's Syndrome, and "known history of rheumatic diseases" for normal samples.
Established Reference Materials: CDC controls and well-characterized in-house sera were used for reproducibility.

8. Sample Size for the Training Set

The document describes studies for validating the device's performance, but it does not mention a "training set" in the context of an AI/machine learning model. The HEp-2000® cell line itself is "transfected" to overexpress the SS-A/Ro antigen, but this is a biological modification, not an algorithmic training process with a data set.

9. How Ground Truth for the Training Set Was Established

As there is no "training set" for an AI algorithm, this question is not applicable to the described device. The "ground truth" equivalent for the development of the HEp-2000® cell line would be the biological understanding of how the transfected cells respond to SS-A/Ro antibodies compared to non-transfected cells, which is based on established scientific principles in immunology and cell biology.

Summary

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DEC 1 8 1998

K983924

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510(k) SUMMARY*

Date Prepared: August 10, 1998 Contact Person: Eric S. Hoy, Ph.D. Name of Device:

· Trade Name HEp-2000® Colorzyme® ANA-Ro Test System
· Common Name HEp-2000® Colorzyme® ANA-Ro Test System
· Classification Name Antinuclear Antibody (21 CFR 866.5100)

Legally marketed device with which this device has been shown to be equivalent: RELISA® SS-A/SS-B Antibody Test System, K955603

Description:

Intended Use:

This test system is for in vitro diagnostic use for the detection of antinuclear antibodies in human serum, with specific identification of autoantibodies to the SS-A/Ro antigen. This test system is to be used as an aid in the detection of antibodies associated with systemic rheumatic disease. The results from this assay can be used as an aid in the diagnosis of autoimmune diseases.

Summary of Technological Characteristics Compared to the Predicate Device:

This device is an indirect immunoenzyme antibody test for the detection of antinuclear antibodies. This 510(k) submission is an extension of K950979. In this new submission we have shown that the transfected HEp-2000® cell line produces a distinctive pattern that is confirmatory for the presence of antibodies to the SS-A/Ro autoantigen. We have compared the present device to an ELISA assay which is confirmatory for antibodies to the SS-A/Ro autoantigen (K955603) .

Description of Laboratory Data That Indicate Substantial Equivalence: NORMAL SAMPLES

Sera from 500 healthy blood donors, 242 females and 258 males, none of whom had any known history of rheumatic diseases, were tested in parallel using commercially available, non-transfected HEp-2 cells and the HEp-2000® Colorzyme® ANA-Ro Test System. In this population, 36 samples (7.2%) showed positive antinuclear antibody tests at a 1:40 dilution of serum. Patterns of staining were identical on the two substrates for 34 of the 36 positive samples. The two samples that showed differences were both from female patients, and both were confirmed to contain anti-SS-A/Ro antibodies. One of these samples showed a weak fine speckled reaction on the non-transfected HEO-2 cells and the typical "SS-A/Ro" staining on the HEp-2000® Colorzyme® ANA-Ro Test System. The other sample was negative on the non-transfected HEp-2 but showed typical "SS-A/Ro" staining on the HED-2000® Colorzyme® ANA

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Ro Test System. The SS-A/Ro specificity of these two samples was confirmed by ELISA testing and by Western immunoblotting. Samples which were negative on the ANA tests were also negative in the ELISA assay.

SERA FROM PATIENTS WITH ONLY RO/SS-A ANTIBODIES

Sera from 46 patients with SLE or Sjögren's Syndrome were tested using commercially available, non-transfected HEp-2 cells and the HEp-2000® Colorzyme® ANA-Ro Test System. All of these sera were confirmed to contain antibodies to the SS-A/Ro autoantigen by ELISA testing and by Western immunoblotting. No other autoantibodies were detected in any of these samples. Thirty six of these samples (78%) were positive (speckled pattern) with the non-transfected HEp-2 cells, and all 46 (100%) were positive (distinctive Ro/SS-A staining pattern) with the HEp-2000® Colorzyme® ANA-Ro Test System.

SERA FROM PATIENTS WITH AUTOANTIBODIES OTHER THAN SS-A/RO

Serum samples from 230 patients with a variety of rheumatic and non-rheumatic diseases were tested in parallel using commercially available, non-transfected HEp-2 cells and the HEp-2000® Colorzyme® ANA-Ro Test System. A single staining pattern was seen with 120 samples, and mixed patterns were seen with 110 samples. Among the total population of 230 samples, 333 staining patterns were identical on both substrates. Twenty nine samples showed the distinctive "SS-A/Ro" staining pattern on the HEp-2000® Test System. Twenty three of these samples showed speckled patterns on the non-transfected HEp-2 cells. The six discrepant samples (positive on HEp-2000®, but negative on nontransfected HEp-2 cells) all had SS-A/Ro antibodies, as demonstrated by the distinctive "SS-A/Ro" staining pattern, ELISA tests, and Western blot confirmation.

TITER COMPARISONS

Because of the overexpression of the SS-A/Ro autoantigen in the HEp-2000® cells, samples that contain anti-Ro/SS-A antibodies show higher titer values on these cells than the values obtained on non-transfected HEp-2 cells. Since none of the other autoantigens in the HEp-2000® cells are affected by the transfection process, sera with other autoantibody specificities do not show significant titer differences between the transfected HEp-2000® cell line and non-transfected HEp-2 cells.

REPRODUCIBILITY TITER

Ten samples, chosen from CDC controls and other well characterized in-house sera, were run on three different lot numbers of HEp-2000® slides, on three different occasions. In no case did a neqative sample show positive results. All titer values were within one two-fold dilution of the established mean titer value for all samples tested.

CONFIRMATION SS-A/Ro ANTIBODIES OF

In a large rheumatology reference laboratory, serum samples from 349 patients with known positive ANA tests were assayed using the HEp-2000® Colorzyme® In this selected population, 239 samples showed the distinctive Test System. SS-A/Ro staining pattern. Positive ELISA tests for SS-A/Ro antibodies were obtained in 238 (99.68) of these samples. An additional 79 samples showed strong speckled and/or homogeneous patterns, but gave positive ELISA tests for SS-A/Ro antibodies.. Thus, if the distinctive SS-A/Ro pattern is seen, it is confirmatory for the presence of SS-A/Ro antibodies, but the absence of the pattern does not rule out the possible presence of SS-A/Ro antibodies.

In the studies outlined above, we have examined a total of 429 sera that contain SS-A/Ro antibodies as confirmed by ELISA testing and/or Western

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Immunoblots, and which showed the distinctive SS-A/Ro staining pattern on the transfected HEp-2000® cell line. We have also seen samples which contain SS-A/Ro antibodies, but do not display the distinctive SS-A/Ro staining pattern, because high levels of other autoantibodies (usually anti-DNA antibodies or anti-Sm/RNP antibodies) mask the SS-A/Ro pattern. Thus, if the distinctive SS-A/Ro pattern is seen, it is confirmatory for the presence of SS-A/Ro antibodies, but the absence of the pattern does not rule out the possible presence of SS-A/Ro antibodies.

In accordance with 21 CFR 807.92(b)(3), we conclude from these data that the present device is substantially equivalent to the predicate device.

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DEPARTMENT OF HEALTH & HUMAN SERVICES

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

DEC 1 8 1998 Eric S. Hoy, Ph.D., SI(ASCP) Chief Scientific Officer Immuno Concepts, Inc. 2280 Springlake Road, Suite 106 Dallas, Texas 75234

Re: K983924 HEP-2000® ANA-Ro Test System Trade Name: Regulatory Class: II Product Code: DHN Dated: November 3, 1998 November 4, 1998 Received:

Dear Dr. Hoy:

We have reviewed your Section 510(k) notification of intent to market the device referenced above and we 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). You may, therefore, market the device, subject to the general controls provisions of the Act. 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.

If your device is classified (see above) into either class II (Special Controls) or class III (Premarket Approval), it may be subject to such additional controls. Existing major regulations affecting your device can be found in the Code of Federal Requlations, Title 21, Parts 800 to 895. A substantially equivalent determination assumes compliance with the Current Good Manufacturing Practice requirements, as set forth in the Quality System Regulation (QS) for Medical Devices: General regulation (21 CFR Part 820) and that, through periodic QS inspections, the Food and Drug Administration (FDA) will verify such Failure to comply with the GMP regulation may result in assumptions. regulatory action. In addition, FDA may publish further announcements concerning your device in the Federal Reqister. Please note: this response to your premarket notification submission does not affect any obligation you might have under sections 531 through 542 of the Act for devices under the Electronic Product Radiation Control provisions, or other Federal laws or regulations.

Image /page/3/Picture/8 description: The image is a black and white logo for the U.S. Department of Health & Human Services. The logo features a stylized image of three human profiles facing right, stacked on top of each other. The profiles are simple and abstract, with curved lines suggesting the faces. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular fashion around the image.

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Page 2

This letter will allow you to begin marketing your device as described in your 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific advice for your device on our labeling requlation (21 CFR Part 801 and additionally 809.10 for in vitro diagnostic devices), please contact the Office of Compliance at (301) 594-4588. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at (301) 594-4639. Also, please note the regulation entitled, "Misbranding by reference to premarket notification"(21 CFR 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers Assistance at its toll-free number (800) 638-2041 or (301) 443-6597, or at its internet address "http://www.fda.gov/cdrh/dsma/dsmamain.html".

Sincerely yours,

Steven Putman

Steven I. Gutman, M.D., M.B.A. Director Division of Clinical Laboratory Devices Office of Device Evaluation Center for Devices and Radiological Health

Enclosure ..

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	.

510(k) number (if known):

Device Name: HEp-2000® Colorzyme® ANA-Ro Test System

Indications for use:

(PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of Device Evaluation (ODE)

Peter E. Malm

(Division Sign-Off)
Division of Clinical Laboratory Devices
510(k) Number K983924

Prescription Use
(Per 21 CFR 801.109) /

Over-The-Counter Use _________________________________________________________________________________________________________________________________________________________

Regulation Number and Section

§ 866.5100 Antinuclear antibody immunological test system.

(a)
Identification. An antinuclear antibody immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the autoimmune antibodies in serum, other body fluids, and tissues that react with cellular nuclear constituents (molecules present in the nucleus of a cell, such as ribonucleic acid, deoxyribonucleic acid, or nuclear proteins). The measurements aid in the diagnosis of systemic lupus erythematosus (a multisystem autoimmune disease in which antibodies attack the victim's own tissues), hepatitis (a liver disease), rheumatoid arthritis, Sjögren's syndrome (arthritis with inflammation of the eye, eyelid, and salivary glands), and systemic sclerosis (chronic hardening and shrinking of many body tissues).(b)
Classification. Class II (performance standards).