EUROIMMUN EUROLINE ENA PROFILE 9AG (IGG)

{0} 1 # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION MEMORANDUM A. 510(k) Number: k140224 B. Purpose for Submission: New device C. Measurand: IgG antibodies specific for nRNP/Sm, Sm, SS-A, Ro-52, SS-B, Scl-70, Jo-1, CENP B, and Ribosomal P-proteins D. Type of Test: Manual and automated qualitative immunoblot assay E. Applicant: EUROIMMUN US Inc. F. Proprietary and Established Names: EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) G. Regulatory Information: 1. Regulation section: 21CFR§866.5100 – Antinuclear Antibody Immunological Test System 2. Classification: Class II 3. Product code: | EUROIMMUN EUROLINE ENA Profile Autoantibodies | Product Code | | --- | --- | | Anti-nRNP/Sm | LKO, Anti-RNP Antibody, Antigen and Control | | Anti-Sm | LKP, Anti-Sm Antibody, Antigen and Control | {1} | Anti-SS-A | LLL, Extractable Antinuclear antibody, Antigen and Control | | --- | --- | | Anti-Ro-52 | | | Anti-SS-B | | | Anti-Scl-70 | | | Anti-Jo-1 | | | Anti-CENP B | LJM, Antinuclear Antibody (Enzyme-labeled), Antigen, Controls | | Anti-Ribosomal P-proteins | MQA, Anti-Ribosomal P Antibodies | 4. Panel: Immunology (82) H. Intended Use: 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 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. 2. Indication(s) for use: Same as intended use 3. Special conditions for use statement(s): For Prescription use only. 4. Special instrument requirements: CanoScan LiDE Series flatbed scanner using ScanGear and EUROLineScan softwares for automated read (refer to k113439). I. Device Description: The EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) consists of antigen coated line blot strips, a positive control (human IgG; 100x concentrate), alkaline phosphatase-labelled goat anti-human IgG conjugate (10X concentrate), sample buffer (ready-to-use), wash buffer (10X concentrate), Nitro blue tetrazolium {2} chloride/5-Bromo-4-chloro-3-indolylphosphate (NBT/BCIP) substrate solution (ready-to-use), 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) and 510(k) number(s): | EUROIMMUN Anti-nRNP/Sm ELISA (IgG) | k123261 | | --- | --- | | EUROIMMUN Anti-Sm ELISA (IgG) | k123261 | | EUROIMMUN Anti-SS-A ELISA (IgG) | k123261 | | INOVA Quanta Lite™ SS-A 52 ELISA | k063565 | | EUROIMMUN Anti-SS-B ELISA (IgG) | k123261 | | EUROIMMUN Anti-Scl-70 ELISA (IgG) | k123261 | | EUROIMMUN Anti-Jo-1 ELISA (IgG) | k123261 | | EUROIMMUN Anti-Centromeres ELISA (IgG) | k123261 | | EUROIMMUN Anti-Ribosomal P-Proteins ELISA (IgG) | k123261 | 2. Comparison with predicate(s): | Similarities | | | | | --- | --- | --- | --- | | Item | Device | | Predicate(s) | | Intended Use | Qualitative detection of IgG antibodies against 9 different antigens on the same strip: nRNP/Sm, Sm, SS-A, Ro-52, SS-B, Scl-70, Jo-1, CENP B and ribosomal P-proteins in human serum to aid in the diagnosis of autoimmune diseases, systemic sclerosis, poly-/dermatomyositis, mixed connective tissue disease and Sjögren’s syndrome, in conjunction with other laboratory and clinical findings.. | | Same | | Capture antigens | nRNP/Sm | Purified U1-nRNP complex;U1-nRNP contains RNP as well as Sm reactive proteins | Same | | | Sm | Purified Sm antigen | Same | | | SS-A | Purified SS-A antigen | Same | | | Ro-52 | Recombinant Ro-52 antigen | Same | | | SS-B | Purified SS-B antigen | Same | | | Scl-70 | Purified Scl-70 antigen | Same | | | Jo-1 | Purified Jo-1 antigen | Same | | | CENP B | Recombinant centromeres protein B | Same | | | Ribosomal P-proteins | Purified ribosomal P antigen | Same | | Assay format | Qualitative, positive/negative | | Same | {3} | Differences | | | | --- | --- | --- | | Item | Device | Predicate(s) | | Assay configuration | All analytes on the same membrane test strip and tested concurrently | Each analyte is coated on polystyrene microwells and tested individually | | Result readout method | Manual visual readout or automated instrument readout | Instrument readout | | Instrument | A flatbed scanner with EUROLineScan software | Spectrophotometer | | Sample Type | Serum | EUROIMMUN: Serum and plasma (EDTA, Li-heparin, Citrate) INOVA: Same | | Sample dilution | 1:101 | EUROIMMUN: 1:201 INOVA: 1:101 | | Detection antibody | Alkaline phosphatase-labeled anti-human IgG | Horseradish peroxidase-labeled anti-human IgG | | Controls | One positive control | EUROIMMUN: Two controls (positive, negative) INOVA: Three controls (high positive, low positive, negative) | | Substrate | Nitro blue tetrazolium chloride/5-Bromo-4-chloro-3-indolylphosphate | 3, 5',5',5' Tetramethylbenzidine | | Interpretation of results | Positive/negative compared to reaction control card (visual readout) or based on grey scale unit (automated readout) | EUROIMMUN: RU/mL (dsDNA: IU/mL) or ratios INOVA: U/mL | 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) EN 13640:2002: Stability Testing of In Vitro Diagnostic Reagents L. Test Principle: The principle of the EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) is that of an enzyme linked immunosorbent assay (ELISA), using a membrane as the solid phase instead of microtiter wells. Different purified antigens have been coated and applied in easy to read lines (bands) onto a membrane strip. {4} Autoantibodies in patient samples bind to the bands and are detected via a secondary antibody linked to an enzyme. The 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 flatbed 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 color reaction if the incubation was performed correctly and so represents a function test on each single strip. 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 either the control band or the strip incubated with the positive control is negative, test results are invalid and should be repeated. The qualitative results are reported for each individual antibody separately as positive or negative. The intensity of the reaction is not reported but only as a means to distinguish between a positive and negative reaction and not as an indication of disease status. The interpretation of the test results does not include a combined score or diagnosis. ## M. Performance Characteristics (if/when applicable): ### 1. Analytical performance: Test results were evaluated by visual or automated reading of the test strips. For visual evaluation, the reaction intensity of each antigen band on the test strip was compared to the two color bars on the reaction control card that correspond to the intensities just below (negative) and just above (positive) the cut-off. For automated evaluation, the test strips are scanned using a flatbed scanner and evaluated with EUROLineScan with a cut-off of 11 grey scale units. #### a. Precision/Reproducibility: Assay reproducibility was determined by testing 6 or 7 samples that cover the complete range of results (negative, positive and near to cut-off). The intra-assay reproducibility is based on 20 replicates tested in one day using the same kit lot. 5 {5} Intra-assay reproducibility: | Antigen band | Sample characterization | Sample number (n) | Visual evaluation result | | | --- | --- | --- | --- | --- | | | | | % positive | % negative | | nRNP/Sm | pos | 2 | 100 | 0 | | | pos (near to cut-off) | 2 | 100 | 0 | | | neg (near to cut-off) | 2 | 0 | 100 | | | neg | 1 | 0 | 100 | | Sm | pos | 2 | 100 | 0 | | | pos (near to cut-off) | 2 | 100 | 0 | | | neg | 2 | 0 | 100 | | SS-A | pos | 2 | 100 | 0 | | | pos (near to cut-off) | 2 | 100 | 0 | | | neg (near to cut-off) | 1 | 0 | 100 | | | neg | 2 | 0 | 100 | | Ro-52 | pos | 2 | 100 | 0 | | | pos (near to cut-off) | 1 | 100 | 0 | | | pos (near to cut-off) | 1 | 60 | 40 | | | neg | 2 | 0 | 100 | | SS-B | pos | 2 | 100 | 0 | | | pos (near to cut-off) | 1 | 100 | 0 | | | neg (near to cut-off) | 1 | 30 | 70 | | | neg | 2 | 0 | 100 | | Scl-70 | pos | 2 | 100 | 0 | | | pos (near to cut-off) | 1 | 50 | 50 | | | neg (near to cut-off) | 1 | 0 | 100 | | | neg (near to cut-off) | 1 | 45 | 55 | | | neg | 2 | 0 | 100 | | Jo-1 | pos | 2 | 100 | 0 | | | pos (near to cut-off) | 1 | 100 | 0 | | | pos (near to cut-off) | 1 | 85 | 15 | | | neg | 2 | 0 | 100 | | CENP B | pos | 2 | 100 | 0 | | | pos (near to cut-off) | 1 | 80 | 20 | | | pos (near to cut-off) | 1 | 60 | 40 | | | neg | 2 | 0 | 100 | | Ribosomal P-proteins | pos | 2 | 100 | 0 | | | pos (near to cut-off) | 2 | 100 | 0 | | | neg | 2 | 0 | 100 | The inter-assay reproducibility is based on 5 different runs using the same kit lot, each run on a different day with 3 replicates per run performed by the same technician. Evaluation was performed both visually using the reaction control card and automatically using the EUROLineScan software with a cutoff of 11 grey scale units. The results are shown in the table below. {6} Inter-assay reproducibility: | Antigen | Sample Characterization | n | Visual evaluation result | | EUROLineScan result | | | --- | --- | --- | --- | --- | --- | --- | | | | | % pos | % neg | % pos | % neg | | nRNP /Sm | pos | 4 | 100 | 0 | 100 | 0 | | | neg (near to cut-off) | 1 | 0 | 100 | 0 | 100 | | | Neg | 2 | 0 | 100 | 0 | 100 | | Sm | pos | 3 | 100 | 0 | 100 | 0 | | | pos (near to cut-off) | 1 | 100 | 0 | 100 | 0 | | | neg (near to cut-off) | 1 | 0 | 100 | 0 | 100 | | | Neg | 1 | 0 | 100 | 0 | 100 | | SS-A | pos | 2 | 100 | 0 | 100 | 0 | | | pos (near to cut-off) | 2 | 100 | 0 | 100 | 0 | | | neg (near to cut-off) | 1 | 0 | 100 | 0 | 100 | | | Neg | 2 | 0 | 100 | 0 | 100 | | Ro-52 | pos | 2 | 100 | 0 | 100 | 0 | | | pos (near to cut-off) | 2 | 100 | 0 | 100 | 0 | | | Neg | 2 | 0 | 100 | 0 | 100 | | SS-B | pos | 2 | 100 | 0 | 100 | 0 | | | pos (near to cut-off) | 2 | 100 | 0 | 100 | 0 | | | neg | 2 | 0 | 100 | 0 | 100 | | Scl-70 | pos | 2 | 100 | 0 | 100 | 0 | | | pos (near to cut-off) | 2 | 100 | 0 | 100 | 0 | | | neg (near to cut-off) | 1 | 0 | 100 | 0 | 100 | | | neg | 2 | 0 | 100 | 0 | 100 | | Jo-1 | pos | 2 | 100 | 0 | 100 | 0 | | | pos (near to cut-off) | 2 | 100 | 0 | 100 | 0 | | | neg | 2 | 0 | 100 | 0 | 100 | | CENP B | pos | 2 | 100 | 0 | 100 | 0 | | | pos (near to cut-off) | 2 | 100 | 0 | 100 | 0 | | | neg | 2 | 0 | 100 | 0 | 100 | | Ribosomal P-proteins | pos | 2 | 100 | 0 | 100 | 0 | | | pos (near to cut-off) | 2 | 100 | 0 | 100 | 0 | | | neg | 2 | 0 | 100 | 0 | 100 | The lot-to-lot reproducibility was tested in 6 different runs (each run in duplicates) using 3 different kit lots. The reproducibility data showed no positive sample was found negative and vice versa. {7} Inter-lot reproducibility: | Antigen band | Sample Characterization | Sample number (n) | Visual evaluation result | | | --- | --- | --- | --- | --- | | | | | % positive | % negative | | nRNP/Sm | pos | 1 | 100 | 0 | | | neg (near to cut-off) | 1 | 8.3 | 91.7 | | | Neg | 1 | 0 | 100 | | Sm | pos (near to cut-off) | 1 | 91.7 | 8.3 | | | neg (near to cut-off) | 1 | 25 | 75 | | | Neg | 1 | 0 | 100 | | SS-A | pos | 1 | 100 | 0 | | | pos (near to cut-off) | 1 | 83.3 | 16.7 | | | Neg | 1 | 0 | 100 | | Ro-52 | pos | 1 | 100 | 0 | | | pos (near to cut-off) | 1 | 91.7 | 8.3 | | | Neg | 1 | 0 | 100 | | SS-B | pos | 1 | 100 | 0 | | | pos (near to cut-off) | 1 | 100 | 0 | | | Neg | 1 | 0 | 100 | | Scl-70 | pos | 1 | 100 | 0 | | | pos (near to cut-off) | 1 | 100 | 0 | | | Neg | 1 | 0 | 100 | | Jo-1 | pos | 1 | 100 | 0 | | | pos (near to cut-off) | 1 | 100 | 0 | | | Neg | 1 | 0 | 100 | | CENP B | pos | 1 | 100 | 0 | | | pos (near to cut-off) | 1 | 75 | 25 | | | Neg | 1 | 0 | 100 | | Ribosomal P-proteins | pos | 1 | 100 | 0 | | | neg (near to cut-off) | 1 | 25 | 75 | | | Neg | 1 | 0 | 100 | The inter-reader reproducibility of 3 samples that cover the complete range of results (negative, positive and near to cut-off) for each antigen was evaluated in 6 runs, each run performed in duplicates by 2 different technicians. No deviation was observed between the individual readings. b. Linearity/assay reportable range: Not applicable c. Traceability, Stability, Expected values (controls, calibrators, or methods): Traceability: There are no recognized standards or reference materials for these analytes. {8} Controls: Controls are derived from human serum and match pre-specified performance criteria. ## Stability: ### i. Real-time and accelerated stability The stability studies were performed in accordance with EN 13640:2002 (Stability Testing of *In Vitro Diagnostic Reagents*) to demonstrate unopened kit shelf-life stability (from the date of manufacture when stored at recommended temperature 2-8°C) and opened kit shelf-life stability. The acceptance criteria are that results do not differ more than one result level [pos, pos (near the cutoff), neg (near the cutoff) and neg] from the reference run (real-time stability study: day 0; accelerated stability study and transportation simulation: 4°C stored sample). For real-time stability study, three lots of all kit components stored at recommended storage temperature 2-8°C were evaluated at different occasions with three samples per antigen band. Original sealed kits and opened kits were demonstrated to be stable for up to 18 and 12 months, respectively, when stored at 2-8°C. For accelerated study, three lots of all kits components were stored for 7 days at 37°C and evaluated with 3 samples per antigen band. The same lots stored at 4°C were used as reference. Reagents were shown to be stable for 7 days at 37°C. ### ii. Transport stability To simulate transportation of kits from Europe to US, one lot of an example EUROLINE test kit with equal composition and technology as the EUROLINE ENA Profile 9 Ag (IgG) was stored for 7 days at 4°C and -20°C and evaluated with 4 different samples. This simulation showed that no deviation was obtained between the results of the differently stored kits for the different antigen bands. ### d. Detection limit: Not applicable ### e. Analytical specificity: #### i) Endogenous Interference: Interference testing was performed for each antigen using at least 6 samples that cover the complete range of results (negative, positive and near to cut-off) for each antigen. Each sample was spiked with two different levels of endogenous interfering substances, namely hemoglobin, triglycerides and bilirubin. Evaluation of test results 9 {9} was performed visually and no significant interference was observed for concentrations up to 500 mg/dL for hemoglobin, 2000 mg/dL for triglycerides and 40 mg/dL for bilirubin. ii) Cross-reactivity: Analytical cross-reactivity was investigated using the ANA reference panel (n=12) of the Center for Disease Control and Prevention (CDC, Atlanta, USA). The ANA reference panel includes: native DNA, SS-B, RNP/SS-B/SS-A, RNP, Sm, U3-RNP, SS-A, Centromere, Scl-70, Jo-1, PM-Scl, and ribosomal P-proteins. Each reference sample showed the expected reactivity against the EUROLINE ENA antigen bands. f. Assay cut-off: The cut-off is predefined by the visual evaluation. A sample is positive if the respective band is clearly visible. The cut-off for each antigen on the EUROLINE was set to the lowest limit of a clearly visible band. To confirm the assay cut-off, 1118 clinically characterized samples sent in to a clinical laboratory for determination of ANA (338 men and 780 women with an average age of 57 years and age range between 18 and 99 years), as well as 150 control samples (79 men and 71 women with an average age of 37 years and age range: 18 – 67 years) were tested. The results showed that negative and positive results can clearly be discriminated by the assay. 2. Comparison studies: a. Method comparison with predicate device: A comparison study was performed with clinically characterized samples obtained from different sources in Europe and North America. The samples were tested with the EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG) and with the ELISA kits (k123261 or k063565) as the predicate devices. In addition, 5 to 12 artificial samples with antibody concentrations close to the cut-off were created for each antigen by mixing positive samples with negative sample as diluent of the same matrix. The total numbers of samples tested for each antigen differ as shown in the table below. 10 {10} | Antigen | Total test | Gender | Mean age and range | Patient samples | | --- | --- | --- | --- | --- | | nRNP/Sm | 936 | 738 female; 133 male; 65 unknown | 50 (2-91) years; 69 unknown | 225 Systemic lupus erythematosus, 53 Mixed connective tissue diseases and 658 control samples* | | * 211 Systemic sclerosis, 88 Sjögren's syndrome, 164 rheumatoid arthritis, 15 fibromyalgia, 22 gastrointestinal diseases, 10 liver diseases, 23 thyroid diseases, 10 muscle diseases, 11 skin diseases, 10 renal, 25 cancer and 69 infectious diseases | | | | | | Sm | 917 | 715 female; 137 male; 65 unknown | 50 (2-91) years; 69 unknown | 259 Systemic lupus erythematosus and 658 control samples* | | * 211 Systemic sclerosis, 88 Sjögren's syndrome, 164 rheumatoid arthritis, 15 fibromyalgia, 22 gastrointestinal diseases, 10 liver diseases, 23 thyroid diseases, 10 muscle diseases, 11 skin diseases, 10 renal, 25 cancer and 69 infectious diseases | | | | | | SS-A | 1036 | 806 female; 165 male; 65 unknown | 51 (2-91) years; 69 unknown | 229 Systemic lupus erythematosus, 88 Sjögren's syndrome and 658 control samples* | | * 211 Systemic sclerosis, 88 Sjögren's syndrome, 164 rheumatoid arthritis, 15 fibromyalgia, 22 gastrointestinal diseases, 10 liver diseases, 23 thyroid diseases, 10 muscle diseases, 11 skin diseases, 10 renal, 25 cancer and 69 infectious diseases | | | | | | Ro-52 | 276 | 158 female; 53 male; 65 unknown | 44 (2-91) years; 69 unknown | 81 Systemic lupus erythematosus and 195 control samples* | | * 15 fibromyalgia, 22 gastrointestinal diseases, 10 liver diseases, 23 thyroid diseases, 10 muscle diseases, 11 skin diseases, 10 renal, 25 cancer, 69 infectious diseases | | | | | | SS-B | 1036 | 806 female; 165 male; 65 unknown | 51 (2-91) years; 74 unknown | 229 Systemic lupus erythematosus, 88 Sjögren's syndrome and 719 control samples* | | * 211 Systemic sclerosis, 149 polymyositis/dermatomyositis, 164 rheumatoid arthritis, 15 fibromyalgia, 22 gastrointestinal diseases, 10 liver diseases, 23 thyroid diseases, 10 muscle diseases, 11 skin diseases, 10 renal, 25 cancer, 69 infectious diseases. | | | | | | Scl-70 | 663 | 482 female; 111 male; 70 unknown; 5 artificial | 54 (2-91) years; 74 unknown | 211 Systemic sclerosis and 452 control samples* | | * 88 Sjögren's syndrome, 164 rheumatoid arthritis, 15 fibromyalgia, 22 gastrointestinal diseases, 10 liver diseases, 23 thyroid diseases, 10 muscle diseases, 11 skin diseases, 10 renal, 25 cancer, 69 infectious diseases, 5 artificial samples mixed to cut-off range | | | | | | | | mixed, 164 rheumatoid arthritis, 15 fibromyalgia, 22 gastrointestinal diseases, 10 liver diseases, 23 thyroid diseases, 10 muscle diseases, 11 skin diseases, 10 renal, 25 cancer, 69 infectious diseases. | | | | Scl-70 | 663 | 482 female; 111 male; 70 unknown; 5 artificial | 54 (2-91) years; 74 unknown | 211 Systemic sclerosis and 452 control samples* | {11} | Antigen | Total test | Gender | Mean age and range | Patient samples | | --- | --- | --- | --- | --- | | Jo-1 | 626 | 421 female; 130 male; 75 unknown; 10 artificial | 52 (2-91) years; 79 unknown | 169 polymyositis/ dermatomyositis and 457 control samples* | | * 88 Sjögren's syndrome, 164 rheumatoid arthritis, 15 fibromyalgia, 22 gastrointestinal diseases, 10 liver diseases, 23 thyroid diseases, 10 muscle diseases, 11 skin diseases, 10 renal, 25 cancer, 69 infectious diseases, 10 artificial samples mixed to cut-off range. | | | | | | CENP B | 670 | 482 female; 111 male; 77 unknown; 12 artificial | 54 (2-91) years; 81 unknown | 211 Systemic sclerosis and 459 control samples* | | * 88 Sjögren's syndrome, 164 rheumatoid arthritis,15 fibromyalgia, 22 gastrointestinal diseases, 10 liver diseases, 23 thyroid diseases, 10 muscle diseases, 11 skin diseases, 10 renal, 25 cancer, 69 infectious diseases, 12 artificial samples mixed to cut-off range. | | | | | | Ribosomal P-proteins | 615 | 434 female; 89 male; 92 unknown; 2 artificial | 45 (2-91) years; 97 unknown | 273 Systemic lupus erythematosus and 342 control samples* | | * 55 Sjögren's syndrome, 90 rheumatoid arthritis, 15 fibromyalgia, 22 gastrointestinal diseases, 10 liver diseases, 23 thyroid diseases, 10 muscle diseases, 11 skin diseases, 10 renal, 25 cancer, 69 infectious diseases, 2 artificial samples mixed to cut-off range. | | | | | The results and the positive and negative agreement between the device and the predicates are shown in the tables below. $95\%$ C.I.'s were calculated by the exact method. | | Predicate ELISA | | Positive Agreement Negative Agreement Overall Agreement % (95% C.I.) | | | --- | --- | --- | --- | --- | | | | Positive | | Negative | | n=936 | | | | 88.5% (81.5% - 93.6%)100% (99.2% - 100%)98.5% (97.5% - 99.2%) | | nRNP/Sm | positive | 108 | 0 | | | | negative | 14 | 814 | | | | | | | | | n = 917 | | | | 82.9% (66.4% - 93.4%)99.8% (99.2% - 100%)99.1% (98.3% - 99.6%) | | Sm | positive | 29 | 2 | | | | negative | 6 | 880 | | | | | | | | | n = 1036 | | | | 99.5% (97.1% - 100%)97.6% (96.4% - 98.6%)98.1% (97.0% - 98.8%) | | SS-A | positive | 189 | 20 | | | | negative | 1 | 826 | | | | | | | | {12} | | Predicate ELISA | | Positive Agreement Negative Agreement Overall Agreement % (95% C.I.) | | | --- | --- | --- | --- | --- | | | | Positive | | Negative | | n = 276 | | | | 68.4% (54.8% - 80.1%) | | Ro-52 | positive | 39 | 1 | 99.5% (97.5% - 100%) | | | negative | 18 | 218 | 93.1% (89.5% - 95.8%) | | | | | | | | n = 1036 | | | | 100% (94.9% - 100%) | | SS-B | positive | 71 | 32 | 96.7% (95.4% - 97.7%) | | | negative | 0 | 933 | 96.9% (95.7% - 97.9%) | | | | | | | | n = 663 | | | | 96.5% (91.3% - 99.0%) | | Scl-70 | positive | 111 | 8 | 98.5% (97.1% - 99.4%) | | | negative | 4 | 540 | 98.2% (96.9% - 99.1%) | | | | | | | | n = 626 | | | | 100% (92.6% - 100%) | | Jo-1 | positive | 48 | 14 | 97.6% (96.0% - 98.7%) | | | negative | 0 | 564 | 97.8% (96.3% - 98.8%) | | | | | | | | n = 670 | | | | 96.2% (87.0% - 99.5%) | | CENP B | positive | 51 | 5 | 99.2% (98.1% - 99.7%) | | | negative | 2 | 612 | 99.0% (97.9% - 99.6%) | | | | | | | | n = 615 | | | | 84.6% (71.9% - 93.1%) | | Ribosomal P-proteins | positive | 44 | 2 | 99.6% (98.7% - 100%) | | | negative | 8 | 561 | 98.4% (97.0% - 99.2%) | b. Matrix comparison: Serum is the only test matrix. 1. Clinical studies: a. Clinical Sensitivity/Clinical Specificity: In total 1149 clinically characterized samples obtained from different study sites were analyzed by the EUROIMMUN EUROLINE ENA Profile 9 Ag (IgG). {13} | Panel | Source* | n (men, women) | Mean age and range in years | | --- | --- | --- | --- | | Systemic lupus erythematosus | 1 | 210 (14,196) | 41 (16-80) | | Systemic sclerosis (SSc) | 1,2 | 209 (23,188) | 59 (22 - 88) | | Diffuse SSc | 1 | 96 | | | Limited SSc | 1 | 113 | | | Polymyositis/ Dermatomyositis | 2 | 149 (38,111) | 56 (14 - 85) | | Mixed connective tissue diseases | 1,3 | 53 (7,46) | 44 (20 - 69) | | Sjörgren's syndrome | 1 | 88 (4,84) | 52 (19 -78) | | Rheumatoid arthritis | 1 | 164 (43,121) | 54 (19 - 80) | | Fibromyalgia | 4 | 15 (0, 15) | 58 (42 -76) | | Gastrointestinal diseasesa | 4 | 22 (8,7, 7 unknown) | 36 (2 -74); 8 unknown | | Liver diseasesb | 5 | 10 | | | Thyroid diseasesc | 6 | 23 (3,20) | 50 (19 - 73) | | Muscle diseasesd | 4 | 10 (3, 6, 1 unknown) | 62 (31- 91); 1 unknown | | Skin diseasese | 4 | 11 (2,7, 2 unknown) | 61 (40 - 84); 4 unknown | | Renal diseasesf | 4 | 10 (5, 1, 4 unknown) | 54 (34 - 80); 4 unknown | | Cancersg | 7 | 25 | | | Infectious diseasesh | 8 | 69 (20, 33, 16 unknown) | 37 (16 - 62); 17 unknown | | * 1. Prof. Hiepe, Dr. Egerer, University Clinic Charité, Berlin, Germany; 2. Prof. Vencovsky, Institute of Rheumatology, Charles University in Prague, Czech Republic; 3. Dr. Mc Hugh, Royal National Hospital for Rheumatic Diseases, Barth, UK; 4. Rheumatological practice, Germany; 5. Prof. D. Bogdanos, Institute of Liver Studies, School of Medicine, Kings's College Hospital, London, UK; 6. Nuclear medical practice, Germany; 7. University Clinic Charité, Berlin, Germany; 8. Dr. Donglai Ma, EUROIMMUN Canada; Medical Center Laboratory, Germany: University of Lübeck, Germany. a Crohn's disease, (n = 5), Ulcerative colitis (n = 5), Celiac disease (n = 5), Diabetes Type I (n = 5), Reflux (n = 2); b Autoimmune hepatitis (n = 5), Primary biliary liver cirrhosis (n = 5); c Grave's disease (n = 12), Hashimoto's disease (n = 11); d Multiple sclerosis (n = 5), Myasthenia gravis (n = 5); e Autoimmune dermatoses (n = 6), Psoriasis (n = 5); f Renal insufficiency (n = 5), Goodpasture syndrome (n = 5); g Prostate carcinoma (n = 5), Ovarian carcinoma (n = 5), Mammary carcinoma (n = 5), Bronchial carcinoma (n = 5), Cervical carcinoma (n =5); h Borreliosis (n = 30), Hepatitis C virus (n = 15), Hepatitis B virus (n = 2), Herpes simplex virus (n = 4), Rubella (n = 15), Measles (n = 1), Parvovirus (n = 2) | | | | The results of the clinical studies are shown in the following tables: {14} | nRNP/Sm | | | | --- | --- | --- | | Panel | Sensitivity (%) | 95% CI | | Systemic lupus erythematosus | 19.5% (41/210) | 14.4 –25.5% | | Mixed connective tissue diseases | 96.2% (51/53) | 87.0 – 99.5% | | | | | | Panel | Specificity (%) | 95% CI | | Systemic sclerosis | 98.6% (208/211) | 95.9 – 99.7% | | Polymyositis/dermatomyositis | 95.3% (142/149) | 90.6 – 98.1% | | Sjögren’s syndrome | 98.9% (87/88) | 93.8 – 100.0% | | Rheumatoid arthritis | 100% (164/164) | 97.8 – 100% | | Fibromyalgia | 93.3% (14/15) | 68.1 – 99.8% | | Gastrointestinal diseases | 100% (22/22) | 84.6 – 100% | | Liver diseases | 100% (10/10) | 69.2 – 100% | | Thyroid diseases | 100% (23/23) | 85.2 – 100% | | Muscle diseases | 100% (10/10) | 69.2 – 100% | | Skin diseases | 100% (11/11) | 71.5 – 100% | | Renal diseases | 100% (10/10) | 69.2 – 100% | | Cancer | 100% (25/25) | 86.3 – 100% | | Infectious diseases | 100% (69/69) | 94.8 – 100% | | Total | 98.5% (795/807) | 97.4 – 99.2% | | Sm | | | | --- | --- | --- | | Panel | Sensitivity (%) | 95% CI | | Systemic lupus erythematosus | 8.36% (18/210) | 5.2 – 13.2% | | | | | | Panel | Specificity (%) | 95% CI | | Systemic sclerosis | 100% (211/211) | 98.3 – 100% | | Polymyositis/dermatomyositis | 99.3% (148/149) | 96.3 – 98.8% | | Sjögren’s syndrome | 98.8% (87/88) | 93.8 – 100% | | Rheumatoid arthritis | 100% (164/164) | 97.8 – 100% | | Fibromyalgia | 100% (15/15) | 97.8 – 100% | | Gastrointestinal diseases | 100% (22/22) | 84.6 – 100% | | Liver diseases | 100% (10/10) | 69.2 – 100% | | Thyroid diseases | 100% (23/23) | 85.2 – 100% | | Muscle diseases | 100% (10/10) | 69.2 – 100% | | Skin diseases | 100% (11/11) | 71.5 – 100% | | Renal diseases | 100% (10/10) | 69.2 – 100% | | Cancer | 100% (25/25) | 86.3 – 100% | | Infectious diseases | 100% (69/69) | 94.8 – 100% | | Total | 99.4% (855/860) | 98.6 – 99.8% | {15} 16 | SS-A | | | | --- | --- | --- | | Panel | Sensitivity (%) | 95% CI | | Systemic lupus erythematosus | 43.3% (91/210) | 36.5 – 50.3% | | Sjögren’s syndrome | 76.1% (67/88) | 65.9 – 84.6% | | | | | | Panel | Specificity (%) | 95% CI | | Systemic sclerosis | 89.6% (189/211) | 84.6 – 93.3% | | Polymyositis/dermatomyositis | 89.3% (133/149) | 83.1 – 93.7% | | Mixed connective tissue diseases | 92.5% (49/53) | 81.8 – 97.9% | | Rheumatoid arthritis | 95.7% (157/164) | 91.4 – 98.3% | | Fibromyalgia | 93.3% (14/15) | 68.1 – 99.8% | | Gastrointestinal diseases | 100% (22/22) | 84.6 – 100% | | Liver diseases | 100% (10/10) | 69.2 – 100% | | Thyroid diseases | 100% (23/23) | 85.2 – 100% | | Muscle diseases | 100% (10/10) | 69.2 – 100% | | Skin diseases | 100% (11/11) | 71.5 – 100% | | Renal diseases | 100% (10/10) | 69.2 – 100% | | Cancer | 100% (25/25) | 86.3 – 100% | | Infectious diseases | 98.6% (68/69)* | 92.2 –100% | | Total | 93.4% (721/772) | 91.4 – 95.0% | | *Borreliosis | | | | Ro-52 | | | | --- | --- | --- | | Panel | Sensitivity (%) | 95% CI | | Systemic lupus erythematosus | 35.2% (74/210) | 28.8 – 42.1% | | Sjögren’s syndrome | 77.3% (68/88) | 67.1 – 85.5% | | | | | | Panel | Specificity (%) | 95% CI | | Systemic sclerosis | 78.2% (165/211) | 72.0 – 83.6% | | Polymyositis/dermatomyositis | 55.0% (82/149) | 46.7 – 63.2% | | Mixed connective tissue diseases | 81.8% (43/53) | 68.0 – 90.6% | | Rheumatoid arthritis | 94.5% (155/164) | 89.8 – 97.5% | | Fibromyalgia | 93.3% (14/15) | 68.1 – 99.8% | | Gastrointestinal diseases | 95.5% (21/22)* | 77.2 – 99.9% | | Liver diseases | 60% (6/10)** | 26.2 – 87.8% | | Thyroid diseases | 100% (23/23) | 85.2 – 100% | | Muscle diseases | 100% (10/10) | 69.2 – 100% | | Skin diseases | 100% (11/11) | 71.5 – 100% | | Renal diseases | 100% (10/10) | 69.2 – 100% | | Cancer | 96.0% (24/25)^ | 79.6 – 99.9% | | Infectious diseases | 98.6% (68/69)^^ | 92.2 –100% | | Total | 81.9% (632/772) | 79.0 – 84.5% | | * Crohn's disease; ** 4 Autoimmune hepatitis; ^ Ovarian carcinoma; ^^ Hepatitis C virus | | | {16} | SS-B | | | | --- | --- | --- | | Panel | Sensitivity (%) | 95% CI | | Systemic lupus erythematosus | 20.0% (42/210) | 14.8 – 26.1% | | Sjögren’s syndrome | 47.7% (42/88) | 37.0 – 58.6% | | | | | | Panel | Specificity (%) | 95% CI | | Systemic sclerosis | 95.3% (201/211) | 91.5 – 97.7% | | Polymyositis/dermatomyositis | 97.3% (145/149) | 93.3 – 99.3% | | Mixed connective tissue diseases | 100% (53/53) | 93.3 – 100% | | Rheumatoid arthritis | 98.2% (161/164) | 94.7 – 99.6% | | Fibromyalgia | 100% (15/15) | 78.2 – 100% | | Gastrointestinal diseases | 100% (22/22) | 84.6 – 100% | | Liver diseases | 100% (10/10) | 69.2 – 100% | | Thyroid diseases | 100% (23/23) | 85.2 – 100% | | Muscle diseases | 100% (10/10) | 69.2 – 100% | | Skin diseases | 100% (11/11) | 71.5 – 100% | | Renal diseases | 100% (10/10) | 69.2 – 100% | | Cancer | 100% (25/25) | 86.3 – 100% | | Infectious diseases | 100% (69/69) | 94.8 – 100% | | Total | 97.7% (754/772) | 96.3 – 98.6% | | Scl-70 | | | | --- | --- | --- | | Panel | Sensitivity (%) | 95% CI | | Systemic sclerosis (SSc) | 30.6% (64/209) | 24.4 – 37.4% | | Diffuse SSc | 60.4% (58/96) | 49.9 – 70.3 | | Limited SSc | 5.3% (6/113) | 2.0 – 11.2 | | | | | | Panel | Specificity (%) | 95% CI | | Systemic lupus erythematosus | 100% (210/210) | 98.3 – 100% | | Polymyositis/dermatomyositis | 99.3% (148/149) | 96.3 – 100% | | Mixed connective tissue diseases | 100% (53/53) | 93.3 – 100% | | Sjögren’s syndrome | 100% (88/88) | 95.9 – 100% | | Rheumatoid arthritis | 99.4% (163/164) | 96.6 – 100% | | Fibromyalgia | 100% (15/15) | 78.2 – 100% | | Gastrointestinal diseases | 100% (22/22) | 84.6 – 100% | | Liver diseases | 100% (10/10) | 69.2 – 100% | | Thyroid diseases | 100% (23/23) | 85.2 – 100% | | Muscle diseases | 100% (10/10) | 69.2 – 100% | | Skin diseases | 90.9% (10/11)* | 58.7 – 99.8% | | Renal diseases | 100% (10/10) | 69.2 – 100% | | Cancer | 100% (2525) | 86.3 – 100% | | Infectious diseases | 100% (69/69) | 94.8 – 100% | | Total | 99.7% (856/859) | 99.0 – 99.9% | | * Psoriasis | | | {17} | CENP B | | | | --- | --- | --- | | Panel | Sensitivity (%) | 95% CI | | Systemic sclerosis (SSc) | 18.0% (90/209) | 36.3 – 50.1% | | Limited SSc | 74.3% (84/113) | 65.3 – 82.1% | | Diffuse SSc | 6.3% (6/96) | 2.3 – 13.1% | | | | | | Panel | Specificity (%) | 95% CI | | Diffuse SSc | 93.7% (90/96) | 86.9 – 97.7% | | Systemic lupus erythematosus | 98.6% (207/210) | 95.9 – 99.7% | | Polymyositis/dermatomyositis | 97.3% (145/149) | 93.3 – 99.3% | | Mixed connective tissue diseases | 98.1% (52/53) | 89.9 – 100% | | Sjögren’s syndrome | 96.6% (85/88) | 90.4 – 99.3% | | Rheumatoid arthritis | 98.8% (162/164) | 95.7 – 99.9% | | Fibromyalgia | 93.3% (14/15) | 68.1 – 99.8% | | Gastrointestinal diseases | 100% (22/22) | 84.6 – 100% | | Liver diseases | 100% (10/10) | 69.2 – 100% | | Thyroid diseases | 100% (23/23) | 85.2 – 100% | | Muscle diseases | 100% (10/10) | 69.2 – 100% | | Skin diseases | 100% (11/11) | 71.5 – 100% | | Renal diseases | 100% (10/10) | 69.2 – 100% | | Cancer | 96% (24/25)* | 79.6 – 99.9% | | Infectious diseases | 100% (69/69) | 94.8 – 100% | | Total | 99.6% (917/921) | 98.9 – 99.9% | | *Ovarian carcinoma | | | | Jo-1 | | | | --- | --- | --- | | Panel | Sensitivity (%) | 95% CI | | Polymyositis/dermatomyositis | 24.2% (36/149) | 17.5 – 31.8% | | | | | | Panel | Specificity (%) | 95% CI | | Systemic lupus erythematosus | 100% (210/210) | 98.3 – 100% | | Systemic sclerosis | 99.1% (209/211) | 96.6 – 99.9% | | Mixed connective tissue diseases | 100% (53/53) | 93.3 – 100% | | Sjögren’s syndrome | 100% (88/88) | 95.9 – 100% | | Rheumatoid arthritis | 99.4% (163/164) | 96.6 – 100% | | Fibromyalgia | 100% (15/15) | 78.2 – 100% | | Gastrointestinal diseases | 100% (22/22) | 84.6 – 100% | | Liver diseases | 100% (10/10) | 69.2 – 100% | | Thyroid diseases | 100% (23/23) | 85.2 – 100% | | Muscle diseases | 100% (10/10) | 69.2 – 100% | | Skin diseases | 100% (11/11) | 71.5 – 100% | | Renal diseases | 100% (10/10) | 69.2 – 100% | | Cancer | 96% (24/25)* | 79.6 – 99.9% | | Infectious diseases | 100% (69/69) | 94.8 – 100% | | Total | 99.6% (917/921) | 98.9 – 99.9% | {18} | Ribosomal P Proteins | | | | --- | --- | --- | | Panel | Sensitivity (%) | 95% CI | | Systemic lupus erythematosus | 4.8% (10/210) | 2.8 – 8.6% | | | | | | Panel | Specificity (%) | 95% CI | | Systemic sclerosis | 100% (211/211) | 98.3 – 100% | | Polymyositis/dermatomyositis | 100% (149/149) | 97.6 – 100% | | Mixed connective tissue diseases | 98.1% (52/53) | 89.9 – 100% | | Sjögren’s syndrome | 100% (88/88) | 95.9 – 100% | | Rheumatoid arthritis | 100% (164/164) | 97.8 – 100% | | Fibromyalgia | 100% (15/15) | 78.2 – 100% | | Gastrointestinal diseases | 100% (22/22) | 84.6 – 100% | | Liver diseases | 100% (10/10) | 69.2 – 100% | | Thyroid diseases | 100% (23/23) | 85.2 – 100% | | Muscle diseases | 100% (10/10) | 69.2 – 100% | | Skin diseases | 100% (11/11) | 71.5 – 100% | | Renal diseases | 100% (10/10) | 69.2 – 100% | | Cancer | 100% (25/25) | 86.3 – 100% | | Infectious diseases | 100% (69/69) | 94.8 – 100% | | Total | 99.8% (858/860) | 99.2 – 100% | b. Other clinical supportive data (when a. is not applicable): Not applicable. 4. Clinical cut-off: See Assay cut-off. 5. Expected values/Reference range: The levels of autoantibodies against nRMP/Sm, Sm, SS-A, Ro-52, SS-B, Scl-70, Jo-1, CENP B and ribosomal P-proteins were analyzed in a panel of 173 samples from US asymptomatic blood donors of mixed age and sex (79 men, 94 women with a mean age of 38.3 years and age range between 19 and 50 years). The result provided in the table below showed that the $\%$ samples found positive for each autoantibody ranges from 0 to 3.5. It is recommended that each laboratory determine its own normal range based on the population and equipment used. {19} | n=173 | EUROLINE ENA Profile 9 Ag (IgG) | | | --- | --- | --- | | | Positive | % | | nRNP/Sm | 0 | 0 | | Sm | 5 | 2.9 | | SS-A | 6 | 3.5 | | Ro-52 | 6 | 3.5 | | SS-B | 1 | 0.6 | | Scl-70 | 1 | 0.6 | | Jo-1 | 0 | 0 | | CENP B | 1 | 0.6 | | Ribosomal P-proteins | 0 | 0 | # N. Proposed Labeling: The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10. # O. Conclusion: The submitted information in this premarket notification is complete and supports a substantial equivalence decision.