ImmuLisa Enhanced RNA POL III Antibody ELISA

K172078 · Immco Diagnostics, Inc. · NYO · Mar 30, 2018 · Immunology

Device Facts

• Record ID: K172078
• Device Name: ImmuLisa Enhanced RNA POL III Antibody ELISA
• Applicant: Immco Diagnostics, Inc.
• Product Code: NYO · Immunology
• Decision Date: Mar 30, 2018
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.5100
• Device Class: Class 2

Indications for Use

An enzyme linked immunoassay (EUSA) for the qualitative detection of anti-RNA POL III IgG antibodies in human serum as an aid in diagnosis (scleroderma) in conjunction with other laboratory and clinical findings.

Device Story

The ImmuLisa Enhanced RNA POL III Antibody ELISA is a solid-phase immunoassay used in clinical laboratories to detect anti-RNA POL III IgG antibodies in human serum. The device utilizes microwells coated with recombinant RNA POL III antigen. Patient serum, calibrators, and controls are incubated in the wells; specific antibodies bind to the antigen. An enzyme-labeled anti-human IgG conjugate is added, followed by a TMB substrate. A spectrophotometer measures the resulting color change at 450 nm. Results are expressed in ELISA units per milliliter (EU/ml) and reported as positive or negative. The test aids clinicians in diagnosing systemic scleroderma, particularly in identifying patients at risk for diffuse skin involvement and hypertensive renal crisis. It is intended for prescription use.

Clinical Evidence

Clinical performance was evaluated using 281 systemic sclerosis samples and 549 autoimmune/infectious disease controls. Method comparison against the predicate showed 96.2% positive percent agreement and 99.4% negative percent agreement (borderline considered positive). Clinical sensitivity for systemic sclerosis was 23.1% (95% CI 18.4-28.6%) and specificity was 98.2% (95% CI 96.6-99.1%). Precision, linearity, and interference studies were conducted; LoD was 3.2 EU/ml. No significant interference was observed for common substances including hemoglobin, bilirubin, and various medications.

Technological Characteristics

Solid-phase ELISA; 96-well microtiter plates coated with recombinant RNA POL III antigen; HRP-conjugated anti-human IgG detection; TMB substrate; spectrophotometric readout at 450 nm; linear range 3.2–160 EU/ml; qualitative/semi-quantitative output.

Indications for Use

Indicated for the qualitative or semi-quantitative detection of anti-RNA POL III IgG antibodies in human serum as an aid in the diagnosis of systemic scleroderma in conjunction with other laboratory and clinical findings.

Regulatory Classification

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).

Predicate Devices

• INOVA QUANTA Lite RNA POL III ELISA

Submission Summary (Full Text)

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Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

IMMCO Diagnostics, Inc. Mr. Kevin Lawson Chief Regulatory Officer 60 Pineview Dr. Buffalo, New York 14228 March 30, 2018

Re: K172078

Trade/Device Name: ImmuLisa Enhanced RNA POL III Antibody ELISA Regulation Number: 21 CFR 866.5100 Regulation Name: Antinuclear antibody immunological test system Regulatory Class: Class II Product Code: NYO Dated: July 7, 2017 Received: July 10, 2017

Dear Kevin Lawson:

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. 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 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); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR

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Part 820); 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 http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). 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 (http://www.fda.gov/DICE) for more information or contact DICE by email ([e-mail]) or phone [phone] or 301-796-7100).

Sincerely,

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For

Lea Carrington Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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# Indications for Use

510(k) Number (if known) K172078

Device Name ImmuLisa™ Enhanced RNA POL III Antibody ELISA

Indications for Use (Describe)

An enzyme linked immunoassay (ELISA) for the qualitative or semi-quantitative detection of anti-RNA POL III IgG antibodies in human serum as an aid in diagnosis of systemic scleroderma) in conjunction with other laboratory and clinical findings.

| Type of Use (Select one or both, as applicable) | |
|--------------------------------------------------------------------------------|-------------------------------------------------------------------------------|
| <div> <span> Prescription Use (Part 21 CFR 801 Subpart D) </span> </div> | <div> <span> Over-The-Counter Use (21 CFR 801 Subpart C) </span> </div> |

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Image /page/3/Picture/0 description: The image shows the logo for Immco Diagnostics, a Trinity Biotech Company. The logo features a cluster of blue and green circles on the left, followed by the word "immco" in blue, with a small circle above the "o". Below "immco" is the word "DIAGNOSTICS" in green, and below that is the text "A Trinity Biotech Company" in a smaller font size.

## ImmuLisa™ Enhanced RNA POL III Antibody ELISA 510(k) Summary

| Submitter: | Immco Diagnostics, Inc. |
|------------------------------|----------------------------------------------------------------------------|
| Address: | 60 Pineview Dr., Buffalo, NY 14228 |
| Phone Number: | 716-691-0091 ext. 110 |
| Contact: | Kevin Lawson |
| Summary Prepared: | 3-22-2018 |
| Device Name: | ImmuLisa Enhanced™ Anti-RNA POL III Antibody ELISA |
| Common Name: | Anti-RNA POL III Antibody ELISA |
| Product Code: | Autoantibodies, anti-ribonucleic acid polymerase (RNAP) III antibody [NYO] |
| Substantially Equivalent to: | INOVA QUANTA Lite™ RNA POL III ELISA |

General Description: Antinuclear antibodies (ANA) occur in sera of patients with various connective tissue disorders such as systemic lupus erythematosus (SLE), mixed connective tissue disease (MCTD), systemic sclerosis / scleroderma (SSc), polymyositis and Siögren's syndrome. These ANA are directed against nucleding DNA, nucleohistone and various extractable nuclear antigens (ENA) such as RNP, Sm, SS-A (Ro), SS-B (La), Centromerase I), Jo-1 and RNA polymerase III (RNA POL III). These antigens are macromolecular complexes of protein and RNA.

Systemic sclerosis is associated with many autoantibodies. The antibody has been observed in a large percentage of patients with limited cutaneous systemic sclerosis (ICSC) or CREST synaud's phenomenon, esophageal motility abnormalities, sclerodactylia and telangietasia) variant of systemic sclerosis or scleroderma. The anti-Scl-70 antibodies are associated with systemic sclerosis with the risk of diffuse skin involvement. The anti-RNA Pol III antibodies are also predictive of diffuse skin involvement, as well as a predictor for the development of hypertensive renal crisis. Anti-RNA Pol III antibodies are frequently found without the presence of anti-Scl-70 or anti-centromere antibodies in patients with systemic sclerosis.

This test is performed as a solid phase immunoassay. Microwells are coated with recombinant RNA POLIII antigen. Controls, calibrators and patient sera are incubated in the antigen coated wells to allow specific antibodies present in the RNA POL III antigen. Bound antibodies are detected by adding an enzyme labeled anti-human IgG conjugate. Specific enzyme substrate (TMB) is then added and the presence of antibodies is detected by a spectrophotometer at 450 nm. Results are expressed in ELISA units per milliliter (EU/ml) and reported as positive or negative.

Intended Use: An enzyme linked immunoassay (EUSA) for the qualitative detection of anti-RNA POL III IgG antibodies in human serum as an aid in diagnosis (scleroderma) in conjunction with other laboratory and clinical findings.

Similarities and Differences: Both kits use recombinant RNA POL III coated on 96 well plates to detect IgG RNA POL III antibody with HRP anti-human lgG conjugate and TMB substrate. The IMMCO kit utilizes a 5 point calibrator curve with a borderline/indeterminate range of 20-25 EU/ml; while the INOVA kit uses a single as a calibrator and has a cutoff of 20 units with no borderline range.

# Non-clinical Tests:

Method Comparison: Both kits were tested with well-characterized systemic sclerosis subjects and disease controls. Semiquantitative results are presented below. Qualitative results are the borderline considered positive results.

| | Other RNA POL III Ab ELISA | | |
|----------------------------------|----------------------------|----------|-------|
| | Positive | Negative | Total |
| IMMCO<br>RNA POL III Ab<br>ELISA | Positive<br>41 | 1 | 42 |
| | Borderline<br>9 | 1 | 10 |
| | Negative<br>2 | 359 | 361 |
| | Total<br>52 | 361 | 413 |

Borderline considered positive

60 Pineview Drive Buffalo, NY 14228-2120 Toll free [phone]

#### www.immco.com

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Image /page/4/Picture/0 description: The image shows the logo for Immco Diagnostics, a Trinity Biotech Company. The logo features a cluster of blue and green dots on the left, followed by the word "immco" in blue, and the word "DIAGNOSTICS" in green. Below that, the words "A Trinity Biotech Company" are written in a smaller font.

Positive Percent Agreement: 96.2% (95% Cl 85.7% - 99.3%) Negative Percent Agreement: 99.4% (95% CI 97.8% - 99.9%) 99.0% (95% Cl 97.8% - 99.7%) Overall Agreement:

Borderline considered negative

Positive Percent Agreement: 78.8% (95% Cl 64.9% - 88.4%) Negative Percent Agreement: 99.7% (95% Cl 98.2% - 100.0%)

Overall Agreement: 97.1% (95% Cl 95.0% - 98.3%)

### Cross Reactivity

Sets of clinical samples were tested to on the Immulisa™ RNA POLIII EUSA. Populations included systemic sclerosis sera as well as autoimmune and infectious disease controls.

| Population | n | n Positive* | % Positive |
|-------------------------------|-----|-------------|------------|
| Systemic sclerosis (SSC) | 281 | 65 | 23.1% |
| Diffuse cutaneous SSc (dcSSC) | 105 | 61 | 58.1% |
| Limited cutaneous SSc (IcSSC) | 176 | 4 | 2.3% |
| Systemic lupus erythematosus | 40 | 0 | 0.0% |
| Sjögren's syndrome | 41 | 0 | 0.0% |
| Myositis | 40 | 1 | 2.5% |
| Rheumatoid arthritis | 40 | 1 | 2.5% |
| HSV-1 | 20 | 0 | 0.0% |
| HSV-2 | 20 | 0 | 0.0% |
| Lyme | 20 | 0 | 0.0% |
| Toxoplasmosis | 20 | 0 | 0.0% |
| CMV | 20 | 0 | 0.0% |
| Rubella | 20 | 0 | 0.0% |
| Syphilis | 20 | 0 | 0.0% |
| Hepatitis C | 20 | 1 | 5.0% |
| Autoimmune hepatitis | 13 | 0 | 0.0% |
| Acute renal failure | 29 | 0 | 0.0% |
| Breast Cancer | 7 | 1 | 14.3% |
| Colorectal Cancer | 10 | 0 | 0.0% |
| Ovarian Cancer | 10 | 0 | 0.0% |
| Pancreatic Cancer | 10 | 0 | 0.0% |
| Esophageal reflux | 31 | 1 | 3.2% |
| Hypertension | 27 | 1 | 3.7% |
| MCTD | 19 | 1 | 5.3% |
| Morphea | 2 | 0 | 0.0% |
| Pulmonary Hypertension | 29 | 0 | 0.0% |
| Psoriasis | 27 | 3 | 11.1% |
| Raynauds | 13 | 0 | 0.0% |
| Vitiligo | 1 | 0 | 0.0% |

#### Precision

Precision was tested with positive specimens selected throughout the range of the assay. Seven patients were run in duplicate, twice per day for 20 days (n=80 replicates per sample). Assays were run with two different operators / equipment sets.

| S# | Mean<br>EU/ml | Total<br>Imprecision<br>SD | Total<br>Imprecision<br>CV% | Within day<br>SD | Within day<br>CV% | Between days<br>SD | Between days<br>CV% | Within run<br>(Repeatability)<br>SD | Within run<br>(Repeatability)<br>CV% |
|----|---------------|----------------------------|-----------------------------|------------------|-------------------|--------------------|---------------------|-------------------------------------|--------------------------------------|
|----|---------------|----------------------------|-----------------------------|------------------|-------------------|--------------------|---------------------|-------------------------------------|--------------------------------------|

60 Pineview Drive ● Toll free [phone] ● tel. [phone] ● fax [phone]

#### www.immco.com

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Image /page/5/Picture/0 description: The image shows the logo for Immco Diagnostics, a Trinity Biotech Company. The logo features a cluster of blue and green dots on the left, followed by the word "immco" in bold, blue letters. Below "immco" is the word "DIAGNOSTICS" in smaller, green letters, and below that is the text "A Trinity Biotech Company" in a smaller, gray font.

| 1 | 10.0 | 0.7 | 7.3% | 0.7 | 7.3% | 0.0 | 0.0% | 0.5 5.1% | |
|---|-------|-----|------|-----|------|-----|------|----------|------|
| 2 | 18.2 | 0.8 | 4.4% | 0.8 | 4.4% | 0.0 | 0.0% | 0.4 | 2.2% |
| 3 | 19.6 | 0.9 | 4.8% | 0.9 | 4.7% | 0.2 | 1.0% | 0.6 | 2.8% |
| 4 | 22.0 | 1.3 | 5.8% | 1.2 | 5.4% | 0.5 | 2.3% | 1.1 | 4.9% |
| 5 | 46.2 | 3.6 | 7.8% | 3.4 | 7.3% | 1.3 | 2.7% | 2.4 | 5.3% |
| 6 | 82.8 | 5.2 | 6.2% | 5.0 | 6.1% | 1.2 | 1.4% | 3.8 | 4.6% |
| 7 | 159.0 | 9.4 | 5.9% | 9.3 | 5.8% | 1.2 | 0.7% | 5.6 | 3.5% |

## Reproducibility

Qualitative reproducibility was tested with 40 runs of samples in the negative range, ~20% below cutoff, in the moderate positive range of the assay and near the qualitative analysis method. Samples were tested in duplicate twice per day for 20 days by two different operators / equipment sets. Assay results for the cutoff specimen produced 60% qualitative (positive) agreement. The sample that was ~20% above cutoff produced 98% qualitative (positive) agreement. All other specimens produced 100% qualitative agreement.

### Limit of Detection

The limit of detection (LoD) was determined based on 60 replicates of the blank and 10 replicates each of 6 low-level (NHS) samples on kits from two different lots. LoD was determined to be 3.2 EU/ml.

### Linearity and Recovery

Linearity and recovery were tested by diluting positive specimens through the assay range in equidistant dilutions and comparing actual vs. expected results. The linear range of the assay was determined be 3.2 (LoD) – 160 EU/ml. Results are summarized below:

| Test Range<br>(EU/ml) | Slope<br>(95% CI) | Y-intercept<br>(95% CI) | R2 | % recovery |
|-----------------------|---------------------|-------------------------|--------|-------------|
| 2.5 to 33.5 | 1.02 (0.96 to 1.10) | -0.15 (-1.72 to 1.43) | 0.9943 | 96% to 117% |
| 8.0 to 66.1 | 0.98 (0.95 to 1.01) | 1.56 (0.13 to 2.98) | 0.9987 | 97% to 109% |
| 31.8 to 169.5 | 0.99 (0.88 to 1.09) | -2.68 (-14.56 to 9.20) | 0.9890 | 91% to 103% |

### Interference

Interference was studied by mixing sera with known RNA POL III antibody levels with negative serum samples spiked with potential interferents and studying deviation from expected results. No significant interference was demonstrated for the following substances at the levels indicated: hemoglobin (2 g/L), bilirubin (342 umol/L), rheumatoid factor (100 EU/ml), cholesterol (13 mmo/L), triglycerides (37 mmo/L), prednisone (0.84 umol /L), Naproxen (25 mg/M/L, Enalapril (0.86 umol/L, Sildenafil (12.9pmol/L), Amlodipine (245 umol/L), Cyclophosphamide (1437 umol/L), Omeprazole (17.4 umol/L), Metoclopramide (1.5 umol/L), Doxycycline (67.5 umol/L), Bosentan (30g/ml), Tocilizumab (549ug/ml), Mycophenolate mofetil (14.01 ug/ml) and Heparin (3000 U/L).

Clinical Study: Sets of clinical samples were tested on the IMMCO RNA POLIII ELISA as described in the Cross Reactivity section. This included 281 systemic sclerosis samples and 549 autoimmune and infectious disease controls.

Systemic sclerosis Clinical Sensitivity: 23.1% (95% C.I. 18.4-28.6%) Systemic sclerosis Clinical Specificity: 98.2% (95% C.I. 96.6-99.1%)

te samples for these studies were considered positive. NHS were excluded in sensitivity/specificity calculations.

Kein lam

Kevin J. Lawson Chief Regulatory Officer

60 Pineview Drive ● USA Toll free [phone]  •  tel. [phone]  •  fax [phone]

www.immco.com