K Number
K140224
Device Name
EUROIMMUN EUROLINE ENA PROFILE 9AG (IGG)
Manufacturer
Date Cleared
2014-12-09

(314 days)

Product Code
Regulation Number
866.5100
AI/MLSaMDIVD (In Vitro Diagnostic)TherapeuticDiagnosticis PCCP Authorized
Intended Use
The EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) kit is an immune lineblot strip test intended for the qualitative detection of IgG class antibodies against nRNP/Sm, Sm, SS-A, Ro-52, SS-B, Scl-70, Jo-1, CENP B and ribosomal Pproteins in human serum. Detection of these antibodies is used as an aid in the diagnosis of autoimmune diseases such as systemic lupus erythematosus, systemic sclerosis, poly-idermatomyositis, mixed connective tissue disease and Sjögren's syndrome, in conjunction with other laboratory and clinical findings. The EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) test kit is intended to be used in a clinical, reference or hospital laboratory. This kit is not designed for point-of-care testing.
Device Description
The EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) consists of antigen coated test strips, a positive control, alkaline phosphatase-labelled anti-human IgG conjugate, sample buffer concentrate, NBT/BCIP substrate solution, incubation tray and test instruction. Evaluation protocol, reaction control card as well as further accessories for use with EUROLineScan are available separately.
More Information

No
The summary describes a traditional immunoassay kit and does not mention any AI or ML components. While it mentions "accessories for use with EUROLineScan are available separately," the summary itself does not describe the EUROLineScan device or its potential AI/ML capabilities.

No
The device is intended for the qualitative detection of antibodies as an aid in the diagnosis of autoimmune diseases, not for therapeutic purposes.

Yes
The device is described as "intended for the qualitative detection of IgG class antibodies...in human serum," and "Detection of these antibodies is used as an aid in the diagnosis of autoimmune diseases." This clearly indicates its role in diagnosis.

No

The device description explicitly lists physical components such as antigen coated test strips, controls, conjugates, buffers, and substrate solution, indicating it is a hardware-based in vitro diagnostic test kit.

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 kit is "intended for the qualitative detection of IgG class antibodies... in human serum." This is a classic description of an in vitro diagnostic test, as it involves testing a sample taken from the human body outside of the body to diagnose or aid in the diagnosis of a disease.
  • Sample Type: The test uses "human serum," which is a biological sample taken from a patient.
  • Purpose: The detection of these antibodies is used "as an aid in the diagnosis of autoimmune diseases." This directly aligns with the purpose of IVD devices, which are used to provide information for the diagnosis, monitoring, or treatment of diseases.
  • Care Setting: The intended use in a "clinical, reference or hospital laboratory" further supports its classification as an IVD, as these are typical settings where IVD tests are performed.
  • Device Description: The components listed (antigen coated test strips, controls, conjugates, buffers, etc.) are all standard components of IVD kits.

N/A

Intended Use / Indications for Use

The EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) kit is an immune lineblot strip test intended for the qualitative detection of IgG class antibodies against nRNP/Sm, Sm, SS-A, Ro-52, SS-B, Scl-70, Jo-1, CENP B and ribosomal Pproteins in human serum. Detection of these antibodies is used as an aid in the diagnosis of autoimmune diseases such as systemic lupus erythematosus, systemic sclerosis, poly-idermatomyositis, mixed connective tissue disease and Sjögren's syndrome, in conjunction with other laboratory and clinical findings. The EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) test kit is intended to be used in a clinical, reference or hospital laboratory. This kit is not designed for point-of-care testing.

Product codes (comma separated list FDA assigned to the subject device)

LJM

Device Description

The EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) consists of antigen coated test strips, a positive control, alkaline phosphatase-labelled anti-human IgG conjugate, sample buffer concentrate, NBT/BCIP substrate solution, incubation tray and test instruction. Evaluation protocol, reaction control card as well as further accessories for use with EUROLineScan are available separately.

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

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Indicated Patient Age Range

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Intended User / Care Setting

The EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) test kit is intended to be used in a clinical, reference or hospital laboratory.

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)

Analytical Performance (Precision/Reproducibility):

  • Intra-Assay Reproducibility was investigated in 20 replicates on one day using the same kit lot.
  • Inter-Assay Reproducibility was investigated in 5 different runs, each run on a different day with 3 replicates per run, using the same kit lot.
  • Inter-Lot Reproducibility was tested in 6 different runs using different kit lots. Each run was performed in duplicates according to the package insert.
  • Inter-Observer Reproducibility was investigated by determinations of different samples covering the whole range of antigens of the EUROLINE ENA Profile 9 Ag (IgG). The samples were tested in 6 different runs. Each run was performed in duplicates according to the package insert and visually evaluated by two different technicians independently.
  • Key Results: Reproducibility was found to be sufficient as no positive sample was found negative and vice versa.

Analytical Performance (Analytical Specificity - Cross-Reactivity):

  • The reactivity of the EUROLINE ENA Profile 9 Ag (IgG) was verified using the CDC ANA reference panel. All samples were found in line with the CDC characterization.

Analytical Performance (Analytical Specificity - Interference):

  • Different samples were spiked with potential interfering substances in 3 different concentrations. No interference was observed with hemolytic, lipemic or icteric samples for concentrations of up to 500 mg/dl for hemoglobin, 2000 mg/dl for triglycerides and 40 mg/dl for bilirubin.

Analytical Performance (Method Comparison):

  • Study Type: Comparison study with clinically characterized samples.
  • Sample Size:
    • nRNP/Sm: 936 samples
    • Sm: 917 samples
    • SS-A: 1036 samples
    • Ro-52: 276 samples
    • SS-B: 1036 samples
    • Scl-70: 663 samples
    • Jo-1: 626 samples
    • CENP B: 670 samples
    • ribosomal P-proteins: 615 samples
    • Patient samples include SLE, MCTD, SSc, PM/DM, Sjögren's, RA. Control samples include gastrointestinal diseases, liver diseases, thyroid diseases, muscle diseases, skin diseases, renal, cancer, infectious diseases, and artificial samples.
  • Key Results: The results of the study show agreement percentages for each analyte when compared to predicate ELISA kits.
    • nRNP/Sm: Positive Agreement 100.0% (99.2% - 100.0%), Negative Agreement 88.5% (81.5% - 93.6%), Overall Agreement 98.5% (97.5% - 99.2%)
    • Sm: Positive Agreement 99.8% (99.2% - 100.0%), Negative Agreement 82.9% (66.4% - 93.4%), Overall Agreement 99.1% (98.3% - 99.6%)
    • SS-A: Positive Agreement 97.6% (96.4% - 98.6%), Negative Agreement 99.5% (97.1% - 100.0%), Overall Agreement 98.1% (97.0% - 98.8%)
    • Ro-52: Positive Agreement 99.5% (97.5% - 100.0%), Negative Agreement 68.4% (54.8% - 80.1%), Overall Agreement 93.1% (89.5% - 95.8%)
    • SS-B: Positive Agreement 96.7% (95.4% - 97.7%), Negative Agreement 96.9% (95.7% - 97.9%)
    • Scl-70: Positive Agreement 98.5% (97.1% - 99.4%), Negative Agreement 96.5% (91.3% - 99.0%), Overall Agreement 98.2% (96.9% - 99.1%)
    • Jo-1: Positive Agreement 97.6% (96.0% - 98.7%), Negative Agreement 100.0% (92.6% - 100.0%), Overall Agreement 97.8% (96.3% - 98.8%)
    • CENP B: Positive Agreement 99.2% (98.1% - 99.7%), Negative Agreement 96.2% (87.0% - 99.5%), Overall Agreement 99.0% (97.9% - 99.6%)
    • Ribosomal P-proteins: Positive Agreement 99.6% (98.7% - 100.0%), Negative Agreement 84.6% (71.9% - 93.1%), Overall Agreement 98.4% (97.0% - 99.2%)

Clinical Studies (Sensitivity & Specificity):

  • Study Type: Clinical studies in cooperation with different sites.
  • Sample Size: 1279 clinically characterized samples.
    • SLE: 210 samples
    • SSc: 211 samples (Diffuse SSc: 96, Limited SSc: 113)
    • PM/DM: 149 samples
    • MCTD: 53 samples
    • Sjögren's: 88 samples
    • RA: 164 samples
    • Fibromyalgia: 15 samples
    • Gastro-intestinal Diseases: 22 samples
    • Liver Diseases: 10 samples
    • Thyroid Diseases: 23 samples
    • Muscle Diseases: 10 samples
    • Skin Diseases: 11 samples
    • Renal Diseases: 10 samples
    • Cancer: 25 samples
    • Infectious Diseases: 69 samples
  • Key Results: Prevalence of antibodies in different disease panels. Examples:
    • SLE: Anti-nRNP/Sm 19.5%, Anti-Sm 8.6%, Anti-SS-A 43.3%, Anti-Ro-52 35.2%, Anti-SS-B 20.0%, Anti-rib. P-prot. 4.8%.
    • SSc: Anti-Scl-70 53.1%, Anti-CENP B 18.0%.
    • PM/DM: Anti-Jo-1 24.2%, Anti-Ro-52 45.0%.
    • MCTD: Anti-nRNP/Sm 96.2%.
    • Sjögren's: Anti-SS-A 76.1%, Anti-Ro-52 77.3%, Anti-SS-B 47.7%.

Expected Values/Reference Range:

  • Sample Size: 173 samples from US asymptomatic blood donors (79 men, 94 women, mean age 38.3 y, age range 19-50 y).
  • Key Results: The prevalence of the assay in the healthy group was 2.9%.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

See "Summary of Performance Studies" for detailed agreement percentages, which include measures of sensitivity and specificity implicitly. For example, for nRNP/Sm, the Positive Agreement corresponds to the sensitivity, and the Negative Agreement corresponds to the specificity.

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

K123261, K063565

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

K113439

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

Not Found

§ 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).

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Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

December 9, 2014

EUROIMMUN US, INC. C/O MR. MICHAEL LOCKE DIRECTOR OF REGULATORY AFFAIRS 1100 THE AMERICAN ROAD MORRIS PLAINS. NJ 07976

Re: K140224

Trade/Device Name: EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) Regulation Number: 21 CFR §866.5100 Regulation Name: Antinuclear Antibody Immunological Test System Regulatory Class: Class II Product Code: LKO, LKP, LLL, LJM, MQA Dated: November 5, 2014 Received: November 6, 2014

Dear Mr. Locke:

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); medical device reporting (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 Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

1

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled. "Misbranding by reference to premarket notification" (21CFR 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.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely vours.

Maria M. Chan -S

Maria M. Chan, Ph.D. 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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DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration

Indications for Use

510(k) Number (if known) K140224

Device Name

EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) kit

Indications for Use (Describe)

The EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) kit is an immune lineblot strip test intended for the qualitative detection of IgG class antibodies against nRNP/Sm, Sm, SS-A, Ro-52, SS-B, Scl-70, Jo-1, CENP B and ribosomal Pproteins in human serum. Detection of these antibodies is used as an aid in the diagnosis of autoimmune diseases such as systemic lupus erythematosus, systemic sclerosis, poly-idermatomyositis, mixed connective tissue disease and Sjögren's syndrome, in conjunction with other laboratory and clinical findings. The EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) test kit is intended to be used in a clinical, reference or hospital laboratory. This kit is not designed for point-of-care testing.

Type of Use (Select one or both, as applicable)

X Prescription Use (Part 21 CFR 801 Subpart D)

Over-The-Counter Use (21 CFR 801 Subpart C)

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ATTACHMENT 1

510(k) SUBSTANTIAL EQUIVALENCE SAFETY & EFFECTIVENESS SUMMARY

(as required by 21 CFR § 807.92)

A. 510(k) Number:

K140224

B. Purpose for Submission:

New analyte(s)/new device {software previously cleared under EUROIMMUN EUROLINE Profile Autoimmune Liver Disease 8 Ag (IgG) (K113439)]

C. Measurand:

Autoantibodies against nRNP/Sm, Sm, SS-A, Ro-52, SS-B, Scl-70, Jo-1, CENP B, and ribosomal Pproteins

D. Type of Test:

Qualitative Immunoblot (Solid Phase ELISA)

E. Applicant:

EUROIMMUN US INC.

F. Proprietary and Established Names:

EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG)

G. Regulatory Information:

    1. Regulation: 21 CFR 866.5100- Antinuclear Antibody Immunological Test System
    1. Classification: Class II
  • Product code: 3. LJM
    1. Panel: Immunology

H. Intended Use(s):

    1. Intended Use(s):
      The EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) kit is an immune lineblot strip test intended for the qualitative detection of IgG class antibodies against nRNP/Sm. Sm. SS-A. Ro-52. SS-B. Scl-70. Jo-1, CENP B and ribosomal P-proteins in human serum. Detection of these antibodies is used as an aid in the diagnosis of autoimmune diseases such as systemic lupus erythematosus, systemic sclerosis, poly-/dermatomyositis, mixed connective tissue disease and Sjögren's syndrome, in conjunction with other laboratory and clinical findings. The EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) test kit is intended to be used in a clinical, reference or hospital laboratory. This kit is not designed for point-of-care testing.
    1. Indication(s) for Use: Same as Intended Use(s).
    1. Special Conditions for Use Statement(s):

Image /page/3/Picture/27 description: The image shows a close-up of a small, rectangular object with a geometric pattern. The object appears to be a component or part of a larger device, possibly electronic. The pattern consists of small squares arranged in a grid, with alternating colors of green and black. The object is encased in a frame with a notch at the top.

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EUROIMMUN=====================================================================================================================================================================

The EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) test kit is intended to be used in a clinical, reference or hospital laboratory. This kit is not designed for point-of-care testing. For Prescription Use Only.

    1. Special Instrument Requirement(s):
      Not Applicable.

L Device Description:

The EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) consists of antigen coated test strips, a positive control, alkaline phosphatase-labelled anti-human IgG conjugate, sample buffer concentrate, NBT/BCIP substrate solution, incubation tray and test instruction. Evaluation protocol, reaction control card as well as further accessories for use with EUROLineScan are available separately.

J. Substantial Equivalence Information:

    1. Predicate Device Name(s):
      EUROIMMUN and Inova Quanta Lite ELISAs (see table below).
    1. Predicate 510(k) Number(s):

| EUROIMMON
EUROLINE ENA
Profile 9 Ag (IgG)

AntigensPredicate Device(s)510(k) No.
nRNP/SmEUROIMMUN Anti-nRNP/Sm ELISA (IgG)K123261
SmEUROIMMUN Anti-Sm ELISA (IgG)K123261
SS-AEUROIMMUN Anti-SS-A ELISA (IgG)K123261
Ro-52Inova Quanta Lite™ SS-A 52 ELISAK063565
SS-BEUROIMMUN Anti-SS-B ELISA (IgG)K123261
Scl-70EUROIMMUN Anti-Scl-70 ELISA (IgG)K123261
Jo-1EUROIMMUN Anti-Jo-1 ELISA (IgG)K123261
CENP BEUROIMMUN Anti-Centromeres ELISA (IgG)K123261
Ribosomal P-proteinsEUROIMMUN Anti-ribosomal P-proteins ELISA (IgG)K123261
    1. Comparison to Predicate Device(s):

Similarities

ItemNew devicePredicate device(s)
Intended UseQualitative detection of IgG class antibodies against 8 different antigens: nRNP/Sm, Sm, SS-A, Ro-52, SS-B, Scl-70, Jo-1, CENP B and rib. P-proteins in human specimens.Same (when combined)
Reaction PrincipleEnzyme immunoassay: enzyme labeled bound patient antibodies are detected with a chromogenic substrate that is converted to a visible colored product at the reaction site.Same
AntigensnRNP/SmPurified U1-nRNP complex; U1-nRNP contains RNP as well as Sm reactive proteinsSame
SmPurified Sm antigenSame
SS-APurified SS-A antigenSame
Ro-52Recombinant Ro-52 antigenSame
SS-BPurified SS-B antigenSame
Scl-70Purified Scl-70 antigenSame
Jo-1Purified Jo-1 antigenSame
CENP BRecombinant centromere protein BSame
Ribosomal P-proteinsPurified ribosomal P antigenSame

Image /page/4/Picture/15 description: The image shows a close-up of a small, rectangular object with a gray border. Inside the border, there are two square patterns, each composed of four smaller squares arranged in a 2x2 grid. The squares in the grid alternate between green and black colors, creating a checkerboard-like pattern within each of the two larger squares. The object appears to be a component or part of a larger device or system.

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Image /page/5/Picture/0 description: The image shows the word "EUROIMMUN" in black letters. There is a green rectangle with four black dots inside it between the words "EUROIMMUN" and "US". The word "US" is also in black letters. The words and the rectangle are underlined with a black line.

Samples

Differences

Serum

Same

ItemNew devicePredicate device(s)
Assay FormatQualitativeSemi-Quantitative
Technology/
ProcedureStandard ELISA technique (solid phase ELISA): sample
incubation with antigen coated strips, followed by a wash
step, incubation with an anti-human IgG enzyme
conjugate; wash step, incubation with substrate, wash
step, air drying and evaluation.Standard ELISA technique (solid phase
ELISA): sample incubation with antigen
coated wells, followed by a wash step,
incubation with an anti-human IgG
enzyme conjugate; wash step,
incubation with substrate, stopping of
the reaction with stop solution,
photometric reading.
Sample Dilution1:101 DilutionEUROIMMUN: 1:201 dilution
Inova: Serum, 1:101 dilution
ControlsPositive ControlEUROIMMUN: 2 controls (positive,
negative)
Inova: 3 controls (high positive, low
positive, negative)
ConjugateAlkaline phosphatase-labeled anti-human IgGAnti-human IgG labeled with
horseradish peroxidase
SubstrateBCIP/NBTTMB
Reported ResultsPositive/Negative (Qualitative)EUROIMMUN: RU/ml or Ratios

K. Standard/Guidance Document Referenced (if applicable):

Guidance for Industry and FDA Staff: Recommendations for Anti-Nuclear Antibody (ANA) Test System Premarket (510(k)) Submissions (January 22, 2009)

Evaluation of Stability of In Vitro Diagnostic Reagents; Approved Guideline. CLSI document EP25-A. Wayne, PA: Clinical and Laboratory Standards Institute; September 2009.

L. Test Principle:

The procedure follows the EUROLINE technique that was cleared previously via the FDA 510(k) processes under 510(k) No. K113439. The assay can be evaluated visually or by means of the EUROLineScan software which was cleared under the K113439 process.

The EUROLINE uses different purified and recombinant antigens that have been coated and applied in easy to read lines onto a membrane. Antibodies are detected via a secondary antibody linked to an enzyme. The principle of the EUROLINE is that of an enzyme linked immunosorbent assay (ELISA), using a membrane as the solid phase instead of microtiter wells.

Patient samples are diluted 1:101 in sample buffer, 1.5 ml of diluted patient sample are added to the test strip lying in the incubation channel and incubated for 30 minutes at room temperature. After incubation the test strips are washed with wash buffer to remove unbound antibodies and 1.5 ml of the anti-human IgG enzyme conjugate reagent is added to each channel. After an additional 30-minutes incubation at room temperature, the test strips are again washed with wash buffer to remove any unbound enzyme conjugate and 1.5 ml of the substrate solution is added. The strips are incubated for 10 minutes at room temperature and then aspirated and washed with dist. water. The test strips can be evaluated visually by comparison of the band intensity with the reaction control card or they can be digitized by use of a flatbet scanner and evaluated with the computer software EUROLineScan.

The control band on the strips contains (non-specific) anti-human IgG, which reacts with the sample IgG to show a strong color reaction if the incubation was performed correctly and so represents a function test on each single strip. If the control band is negative, the test is invalid and should be repeated.

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EUROIMMUN---------------------------------------------------------------------------------------------------------------------------------------------------------------------

The positive control contains a mixture of the targeted antibodies which bind to the antigen coated on the blot strips. A strip incubated with the positive control shows a positive result. If the positive control is negative, the test results are invalid and should be repeated.

The qualitative results are reported for each individual antibody separately. The interpretation of the test results does not include a combined score or diagnosis.

M. Performance Characteristics (where applicable):

1. Analytical Performance:

  • Precision/Reproducibility a.
    Reproducibility was investigated by repeated determinations of different samples covering the whole range of antigens of the EUROLINE ENA Profile 9 Ag (IgG).

  • · Intra-Assay Reproducibility was investigated in 20 replicates on one day using the same kit lot.

  • · Inter-Assay Reproducibility was investigated in 5 different runs, each run on a different day with 3 replicates per run, using the same kit lot.

  • · Inter-Lot Reproducibility was tested in 6 different runs using different kit lots. Each run was performed in duplicates according to the package insert.

  • · Inter-Observer Reproducibility was investigated by determinations of different samples covering the whole range of antigens of the EUROLINE ENA Profile 9 Ag (IgG). The samples were tested in 6 different runs. Each run was performed in duplicates according to the package insert and visually evaluated by two different technicians independently.

Reproducibility was found to be sufficient as no positive sample was found negative and vice versa.

  • b. Linearity/Assay Reportable Range:
    Not Applicable.

  • C. Traceability, Stability, Expected Values (Controls, Calibrators or Methods):
    A recognized standard or reference material for antibodies against nRNP/Sm. Sm, SS-A, Ro-52, SS-B, Scl-70, Jo-1, CENP B, dsDNA, nucleosomes, histones and ribosomal P-proteins is not available.

Stability studies were conducted in accordance with the international standard DIN EN 13640:2002: Stability testing of in vitro diagnostics reagents (which was aligned by CLSI EP25-A). A real-time stability study was performed on original sealed and opened products to establish the shelf life claim and opened reagent stability claims. An accelerated stability study was performed for preliminary claims and transportation simulation at elevated temperatures. Another transportation under freeze conditions was also performed. Three production lots of the EUROLINE ENA Profile 9 Ag (IgG) and an additional production lot for testing under "freeze" conditions were tested with three samples per antigen band. Acceptance criterium was that the results do not differ more than one result level from the reference run (real-time study: day 0; accelerated study and transportation simulation: 4°C stored sample). The studies support a minimum shelf life claim of 18 months for original sealed components and a stability claim of 12 months for opened components.

  • Detection Limit: d.
    Not Applicable.

  • Analytical Specificity: e.
    Cross-Reactivity: The reactivity of the EUROLINE ENA Profile 9 Ag (IgG) was verified using the CDC ANA reference panel. All samples were found in line with the CDC characterization.

Interference: Different samples were spiked with potential interfering substances in 3 different concentrations. No interference was observed with hemolytic, lipemic or icteric samples for

Image /page/6/Picture/23 description: The image shows a close-up of a small, rectangular object with a symmetrical design. The object appears to have a border or frame around a central area containing two square sections. Each square section is divided into a 2x2 grid, with alternating colors of green and black. The arrangement of the green and black squares is different in each of the two sections.

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EUROIMMUN-

concentrations of up to 500 mg/dl for haemoglobin, 2000 mg/dl for triglycerides and 40 mg/dl for bilirubin.

  • f. Assay Cut-off:
    The cut-off intensity is defined as the lowest limit of a clearly visible band.

    1. Analytical Performance:
    • a. Method Comparison

A comparison study was performed with clinically characterized samples obtained from different sources. The samples were tested with the EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) and with the ELISA kits (k123261or K063565) as the predicate devices. For each parameter the same clinical disease groups were included as for the respective predicate ELISA. The results of the study are shown in the tables below. 95% C.I.'s were calculated by the exact method.

n = 936*PredicateNegative / Positive / Overall Agreement
positivenegative% (95% C.I.)
nRNP/Smpositive1080100.0% (99.2% - 100.0%)
negative1481488.5% (81.5% - 93.6%)
98.5% (97.5% - 99.2%)

Patient samples: 225 SLE, 53 MCTD; Control samples; 22 gastrointestinal diseases, 10 liver diseases, 10 liver diseases, 23 thyroid diseases, 10 muscle diseases, 11 skin diseases, 10 renal, 25 cancer, 69 infectious diseases

Sex: 133 men, 738 women, 65 unknown; Age: mean 50 y, range 2 to 91 y, 69 unknown
--------------------------------------------------------------------------------------------------
n = 917*PredicateNegative / Positive / Overall Agreement
positivenegative% (95% C.I.)
Smpositive29299.8% (99.2% - 100.0%)
negative688082.9% (66.4% - 93.4%)
99.1% (98.3% - 99.6%)

Patient samples: 259 SLE; Control samples: 211 SSc, 88 SS, 164 RA, 15 fibromyalgia, 22 gastrointestinal diseases, 23 thyroid diseases, 10 muscle diseases, 11 skin diseases, 10 renal, 25 cancer, 69 infectious diseases

Sex: 137 men, 715 women, 65 unknown; Age: mean 50 y, range 2 to 91 y, 69 unknown

n = 1036*PredicateNegative / Positive / Overall Agreement
SS-Apositivenegative% (95% C.I.)
positive1892097.6% (96.4% - 98.6%)
negative182699.5% (97.1% - 100.0%)
98.1% (97.0% - 98.8%)

Patient samples: 229 SLE, 88 SS; Control samples: 211 SSc, 15 fbromyalgia, 22 gastrointestinal diseases, 10 liver diseases, 23 thyroid diseases. 10 muscle diseases. 11 skin diseases. 10 renal. 25 cancer. 69 infectious diseases Sex: 165 men, 806 women, 65 unknown; Age: mean 51 y, range 2 to 91 y, 69 unknown

n = 276*PredicateNegative / Positive / Overall Agreement
positivenegative% (95% C.I.)
Ro-52positive39199.5% (97.5% - 100.0%)
negative1821868.4% (54.8% - 80.1%)
93.1% (89.5% - 95.8%)

Patient samples: 81 SLE; Control samples: 12 fibromyalgia, 22 gastrointestinal diseases, 23 thyroid diseases, 10 muscle diseases, 11 skin diseases, 10 renal, 25 cancer, 69 infectious diseases

Sex: 53 men, 158 women, 65 unknown; Age: mean 44 y, range 2 to 91 y, 69 unknown

n = 1036*PredicateNegative / Positive / Overall Agreement
positivenegative% (95% C.I.)
SS-Bpositive713296.7% (95.4% - 97.7%)
negative093396.9% (95.7% - 97.9%)

Patient samples: 229 SLE, 88 SS; Control samples: 211 SSc, 15 fibromyalgia, 22 gastrointesinal diseases, 23 thyroid diseases, 10 muscle diseases, 11 skin diseases, 10 renal, 25 cancer, 69 infectious diseases

Image /page/7/Picture/21 description: The image shows a close-up of a graphic with two square shapes inside of a larger rectangular shape. Each square is divided into four smaller squares, with a checkerboard pattern of green and black. The squares are positioned side by side within the larger shape, which has a thick, dark outline.

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EUROIMMUN

Sex: 165 men, 806 women, 65 unknown; Age: mean 51 y, range 2 to 91 y, 69 unknown

n = 663*PredicateNegative / Positive / Overall Agreement
positivenegative% (95% C.I.)
Scl-70positive111898.5% (97.1% - 99.4%)
negative454096.5% (91.3% - 99.0%)
98.2% (96.9% - 99.1%)

Patient samples: 211 SSc; Control samples, 22 gastrointestinal diseases, 10 liver diseases, 10 liver diseases, 10 muscle diseases, 11 skin diseases, 10 renal, 25 cancer, 69 infectious diseases, 5 artificial samples mixed to cut-off range Sex: 111 men, 482 women, 70 unknown, 5 artificial; Age: mean 54 y, range 2 to 91 y, 74 unknown

n = 626*PredicateNegative / Positive / Overall Agreement
positivenegative% (95% C.I.)
Jo-1positive481497.6% (96.0% - 98.7%)
negative0564100.0% (92.6% - 100.0%)
97.8% (96.3% - 98.8%)

Petient samples: 169 PM/DM; Control samples; 22 gastrointestinal diseases, 10 liver diseases; 20 thyroid diseases, 10 muscle diseases, 11 skin diseases, 10 renal, 25 cancer, 69 infectious diseases, 10 artificial samples mixed to cut-off range Sex: 130 men, 421 women, 75 unknown; Age: mean 52 y, range 2 to 91 y, 79 unknown

n = 670*PredicateNegative / Positive / Overall Agreement
positivenegative% (95% C.I.)
CENP Bpositive51599.2% (98.1% - 99.7%)
negative261296.2% (87.0% - 99.5%)
99.0% (97.9% - 99.6%)

Patient samples: 211 SSc; Control samples, 22 gastrointestinal disease, 10 liver diseases, 10 liver diseases, 10 musce diseases, 11 skin diseases, 10 renal, 25 cancer, 69 infectious diseases, 12 artificial samples mixed to cut-off range Sex: 111 men, 482 women, 77 unknown; Age: mean 54 y, range 2 to 91 y, 81 unknown

n = 615*PredicateNegative / Positive / Overall Agreement
ribosomal
P-proteinspositivenegative% (95% C.I.)
positive44299.6% (98.7% - 100.0%)
negative856184.6% (71.9% - 93.1%)
98.4% (97.0% - 99.2%)

Patient samples: 273 SLE; Control samples, 22 gastrointestinal diseases, 10 liver diseases, 10 hver diseases, 10 muscle diseases, 11 skin diseases, 10 renal, 25 cancer, 69 infectious diseases, 2 artificial samples mixed to cut-off range Sex: 89 men, 434 women, 92 unknown; Age: mean 45 y, range 2 to 91 y, 97 unknown

  • b. Matrix Comparison:
    Not Applicable.

3. Clinical Studies:

  • Sensitivity & Specificity: a.
    Clinical studies were performed in cooperation with different sites (see below). In total 1279 clinically characterized samples were investigated for antibodies against nRNP/Sm, Sm, SS-A, Ro-52, SS-B, Scl-70, Jo-1, CENP B and ribosomal P-proteins (IgG). The results obtained with the EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) are shown in the tables below. 95% C.I. are calculated by the exact method.

Image /page/8/Picture/16 description: The image shows a close-up of a small, rectangular object with a black outline. Inside the object are two square shapes, each divided into four smaller squares. The squares are colored in a checkerboard pattern of green and black. The object appears to be a component or part of a larger device or system.

9

EUROIMMUNฐ‎ ———— US

EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG)

| No. | Panel | n
(men, women, unknown) | Mean age
(age range, unknown) | Anti-nRNP/ Sm | Anti-Sm | Anti-SS-A | Anti-Ro-52 | Anti-SS-B | Anti-Scl-70 | Anti-Jo-1 | Anti-CENP B | Anti-rib. P-prot. |
|-----|----------------------------|----------------------------|----------------------------------|------------------------------|----------------------------|------------------------------|------------------------------|------------------------------|-------------------------------|------------------------------|------------------------------|---------------------------|
| 1 | SLE | 210
(14, 196) | 41 y
(16 – 80 y) | 41 (19.5%)
(14.4 - 25.5%) | 18 (8.6%)
(5.2 - 13.2%) | 91 (43.3%)
(36.5 - 50.3%) | 74 (35.2%)
(28.8 - 42.1%) | 42 (20.0%)
(14.8 - 26.1%) | 0 (0.0%)
(0.0 - 1.7%) | 0 (0.0%)
(0.0 - 1.7%) | 3 (1.4%)
(0.3 - 4.1%) | 10 (4.8%)
(2.3 - 8.6%) |
| 2 | SSc | 211
(23, 188) | 59 y
(22 – 88 y) | 3 (1.4%)
(0.3 - 4.1%) | 0 (0.0%)
(0.0 - 1.7%) | 22 (10.4%)
(6.7 - 15.4%) | 46 (21.8%)
(16.4 - 28.0%) | 10 (4.7%)
(2.3 - 8.5%) | 112 (53.1%)
(46.1 - 60.0%) | 2 (0.9%)
(0.1 - 3.4%) | 38 (18.0%)
(13.1 - 23.9%) | 0 (0.0%)
(0.0 - 1.7%) |
| 3a | Diffuse SSc | 96 | | | | | | | 58 (60.4%)
(49.9 - 70.3%) | | 6 (6.3%)
(2.3 - 13.1%) | |
| 3b | Limited SSc | 113 | | | | | | | 6 (5.3%)
(2.0 - 11.2%) | | 84 (74.3%)
(65.3 - 82.1%) | |
| 4 | PM/DM | 149
(38, 111) | 56 y
(14 – 85 y) | 7 (4.7%)
(1.9 - 9.4%) | 1 (0.7%)
(0.0 - 3.7%) | 16 (10.7%)
(6.3 - 16.9%) | 67 (45.0%)
(36.8 - 53.3%) | 4 (2.7%)
(0.7 - 6.7%) | 1 (0.7%)
(0.0 - 3.7%) | 36 (24.2%)
(17.5 - 31.8%) | 4 (2.7%)
(0.7 - 6.7%) | 0 (0.0%)
(0.0 - 2.4%) |
| 5 | MCTD | 53
(7, 46) | 44 y
(20 – 69 y) | 51 (96.2%)
(87.0 - 99.5%) | 3 (5.7%)
(1.2 - 15.7%) | 4 (7.5%)
(2.1 - 18.2%) | 10 (18.9%)
(9.4 - 32.0%) | 0 (0.0%)
(0.0 - 6.7%) | 0 (0.0%)
(0.0 - 6.7%) | 0 (0.0%)
(0.0 - 6.7%) | 1 (1.9%)
(0.0 - 10.1%) | 1 (1.9%)
(0.0 - 10.1%) |
| 6 | Sjögren's | 88
(4, 84) | 52 y
(19 – 78 y) | 1 (1.1%)
(0.0 - 6.2%) | 1 (1.1%)
(0.0 - 6.2%) | 67 (76.1%)
(65.9 - 84.6%) | 68 (77.3%)
(67.1 - 85.5%) | 42 (47.7%)
(37.0 - 58.6%) | 0 (0.0%)
(0.0 - 4.1%) | 0 (0.0%)
(0.0 - 4.1%) | 3 (3.4%)
(0.7 - 9.6%) | 0 (0.0%)
(0.0 - 4.1%) |
| 7 | RA | 164
(43, 121) | 54 y
(19 – 80 y) | 0 (0.0%)
(0.0 - 2.2%) | 0 (0.0%)
(0.0 - 2.2%) | 7 (4.3%)
(1.7 - 8.6%) | 9 (5.5%)
(2.5 - 10.2%) | 3 (1.8%)
(0.4 - 5.3%) | 1 (0.6%)
(0.0 - 3.4%) | 1 (0.6%)
(0.0 - 3.4%) | 2 (1.2%)
(0.1 - 4.3%) | 0 (0.0%)
(0.0 - 2.2%) |
| No. | Panel | n
(men, women, unknown) | Mean age
(age range, unknown) | Anti-nRNP/ Sm | Anti-Sm | Anti-SS-A | Anti-Ro-52 | Anti-SS-B | Anti-Scl-70 | Anti-Jo-1 | Anti-CENP B | Anti-rib. P-prot. |
| 8 | Fibromyalgia | 15
(0, 15) | 58 y
(42 – 76 y) | 1 (6.7%)
(0.2 – 31.9%) | 0 (0.0%)
(0.0 – 21.8%) | 1 (6.7%)
(0.2 – 31.9%) | 1 (6.7%)
(0.2 – 31.9%) | 0 (0.0%)
(0.0 – 21.8%) | 0 (0.0%)
(0.0 – 21.8%) | 0 (0.0%)
(0.0 – 21.8%) | 1 (6.7%)
(0.2 – 31.9%) | 0 (0.0%)
(0.0 – 21.8%) |
| 9 | Gastro-intestinal Diseases | 22
(8, 7, 7) | 36 y
(2 – 74 y, 8) | 0 (0.0%)
(0.0 – 15.4%) | 0 (0.0%)
(0.0 – 15.4%) | 0 (0.0%)
(0.0 – 15.4%) | 1 (4.5%)
(0.1 – 22.8%) | 0 (0.0%)
(0.0 – 15.4%) | 0 (0.0%)
(0.0 – 15.4%) | 0 (0.0%)
(0.0 – 15.4%) | 0 (0.0%)
(0.0 – 15.4%) | 0 (0.0%)
(0.0 – 15.4%) |
| 10 | Liver Diseases | 10
(3, 20) | 50 y
(19 – 73 y) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) | 4 (40.0%)
(12.2 – 73.8%) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) |
| 11 | Thyroid Diseases | 23
(3, 20) | 50 y
(19 – 73 y) | 0 (0.0%)
(0.0 – 14.8%) | 0 (0.0%)
(0.0 – 14.8%) | 0 (0.0%)
(0.0 – 14.8%) | 0 (0.0%)
(0.0 – 14.8%) | 0 (0.0%)
(0.0 – 14.8%) | 0 (0.0%)
(0.0 – 14.8%) | 0 (0.0%)
(0.0 – 14.8%) | 0 (0.0%)
(0.0 – 14.8%) | 0 (0.0%)
(0.0 – 14.8%) |
| 12 | Muscle Diseases | 10
(3, 6, 1) | 62 y
(31 – 91 y, 1) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) |
| 13 | Skin Diseases | 11
(2, 7, 2) | 61 y
(40 – 84 y, 4) | 0 (0.0%)
(0.0 – 28.5%) | 0 (0.0%)
(0.0 – 28.5%) | 0 (0.0%)
(0.0 – 28.5%) | 0 (0.0%)
(0.0 – 28.5%) | 0 (0.0%)
(0.0 – 28.5%) | 1 (9.1%)
(0.2 – 41.3%) | 0 (0.0%)
(0.0 – 28.5%) | 0 (0.0%)
(0.0 – 28.5%) | 0 (0.0%)
(0.0 – 28.5%) |
| 14 | Renal Diseases | 10
(5, 1, 4) | 54 y
(34 – 80 y, 4) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) | 0 (0.0%)
(0.0 – 30.8%) |
| 15 | Cancer | 25 | | 0 (0.0%)
(0.0 – 13.7%) | 0 (0.0%)
(0.0 – 13.7%) | 0 (0.0%)
(0.0 – 13.7%) | 1 (4.0%)
(0.1 – 20.4%) | 0 (0.0%)
(0.0 – 13.7%) | 0 (0.0%)
(0.0 – 13.7%) | 1 (4.0%)
(0.1 – 20.4%) | 0 (0.0%)
(0.0 – 13.7%) | 0 (0.0%)
(0.0 – 13.7%) |
| No. | Panel | n
(men, women, unknown) | Mean age
(age range, unknown) | Anti-nRNP/ Sm | Anti-Sm | Anti-SS-A | Anti-Ro-52 | Anti-SS-B | Anti-Scl-70 | Anti-Jo-1 | Anti-CENP B | Anti-rib. P-prot. |
| 16 | Infectious
Diseases | 69
(20, 33, 16) | 37 y
(16 – 62 y, 17) | 0 (0.0%)
(0.0–5.2%) | 0 (0.0%)
(0.0–5.2%) | 1 (1.4%)
(0.0–7.8%) | 1 (1.4%)
(0.0–7.8%) | 0 (0.0%)
(0.0–5.2%) | 0 (0.0%)
(0.0–5.2%) | 0 (0.0%)
(0.0–5.2%) | 0 (0.0%)
(0.0–5.2%) | 0 (0.0%)
(0.0–5.2%) |

Image /page/9/Picture/3 description: The image shows a diagram of a rectangular object with two square grids inside. Each grid is divided into four smaller squares, with alternating green and black colors. The top and left squares are green, while the bottom and right squares are black. The object has a gray outline and a small notch at the top.

10

EUROIMMUNฐ‎ ———— US

EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG)

Image /page/10/Picture/3 description: The image shows a diagram of a cartridge with two square patterns inside. Each square pattern is divided into four smaller squares, with alternating colors of green and black. The cartridge is outlined in gray and has a small notch at the top.

11

EUROIMMI

EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG)

b. Other Clinical Supportive Data (when a. and b. are not applicable): Not Applicable.

4. Clinical Cut-off:

See Assay Cut-off.

5. Expected Values/Reference Range:

To establish the reference range of antibodies against nRNP/Sm, Sm, SS-A, Ro-52, SS-B, Sc1-70, Jo-1, CENP B and ribosomal P-proteins (IgG), 173 samples from US asymptomatic blood donors (79 men, 94 women, mean age 38.3 y, age range 19-50 y) were tested with the EUROLINE ENA Profile 9 Ag (IgG). The prevalence of the assay in the healthy group was 2.9%.

N. Performance Characteristics (where applicable):

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

O. Performance Characteristics (where applicable):

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

Name/Title

12.1.2014

Date

Michael Locke/Dir. Of Regulatory Affairs

Michael A. Locke
Signature

Image /page/11/Picture/15 description: The image shows a schematic diagram of an electrical component or connector. The diagram depicts a rectangular outline with two sets of four squares inside. The squares are colored in a checkerboard pattern, with one set having green and black squares, and the other set having black and green squares.