EliA anti-TG Immunoassay, EliA anti-TPO Immunoassay, EliA Thyroid Positive Control 250, EliA Thyroid Positive Control 2500/5000

{0} 1 # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION MEMORANDUM A. 510(k) Number: K151799 B. Purpose for Submission: New device C. Measurand: IgG Antibodies to TG (Thyroglobulin) and TPO (Thyroid peroxidase) D. Type of Test: Fully automated, quantitative, Immunofluorescence E. Applicant: Phadia AB, Sweden F. Proprietary and Established Names: EliA™ anti-TG Immunoassay EliA™ anti-TPO Immunoassay EliA™ Thyroid Positive Control 250 EliA™ Thyroid Positive Control 2500/5000 G. Regulatory Information: 1. Regulation section: 21 CFR § 866.5870: Thyroid autoantibody immunological test system 21 CFR § 862.1660: Quality control material (assayed and unassayed) 2. Classification: Class II (Assay) Class I (Controls) 3. Product code: JZO: System, Test, Thyroid Autoantibody JJY: Multi-Analyte Controls, All Kinds (assayed) {1} 4. Panel: Immunology (82) H. Intended Use: 1. Intended use(s): a. EliA™ anti-TG Immunoassay EliA™ anti-TG is intended for the in vitro quantitative measurement of IgG antibodies directed to thyroglobulin (TG) in human serum and plasma (Li-heparin, EDTA) as an aid in the clinical diagnosis of autoimmune thyroiditis and Graves' disease in conjunction with other laboratory and clinical findings. EliA anti-TG uses the EliA IgG method on the instruments Phadia 250 and Phadia 2500/5000. b. EliA™ anti-TPO Immunoassay EliA™ anti-TPO is intended for the in vitro quantitative measurement of IgG antibodies directed to thyroid peroxidase (TPO) in human serum and plasma (Li-heparin, EDTA,) as an aid in the clinical diagnosis of autoimmune thyroiditis and Graves' disease in conjunction with other laboratory and clinical findings. EliA anti-TPO uses the EliA IgG method on the instruments Phadia 250 and Phadia 2500/5000. c. EliA™ Thyroid Positive Control 250 and 2500/5000 EliA™ Thyroid Positive Control 250 and EliA Thyroid Positive Control 2500/5000 is intended for laboratory use in monitoring the performance of in vitro measurement of antibodies against thyroid peroxidase (TPO) and thyroglobulin (TG) with Phadia 250 and Phadia 2500/5000 using the EliA IgG method. d. EliA™ Thyroid Negative Control 250 and 2500/5000 EliA™ IgG/IgM/IgA Negative Control 250 and EliA™ IgG/IgM/IgA Negative Control 2500/5000 is intended for laboratory use in monitoring the performance of in vitro measurement of autoantibodies with Phadia 250 and Phadia 2500/5000 using the EliA™ IgG or IgM or IgA method. 2. Indication(s) for use: Same as above 3. Special conditions for use statement(s): For prescription use only 4. Special instrument requirements: For use on the Phadia® 250 or Phadia® 2500/5000 instruments 2 {2} I. Device Description: The new devices belong to a fully integrated and automated system for immunodiagnostic testing. It comprises a Fluorescence-Immunoassay test system using EliA single wells as the solid phase and is intended to be performed on the Phadia 250 and Phadia 2500/5000 instruments. The EliA™ reagents are available as modular packages, each purchased separately. All packages except the positive and negative controls are required to carry out an EliA™ anti-TG or anti-TPO test. 1. Antibody-Coated Wells a. EliA™ anti-TG wells are coated with a human thyroglobulin antigen – four carriers (16 wells each), ready to use. b. EliA™ anti-TPO wells are coated with a human recombinant thyroid peroxidase antigen – four carriers (16 wells each), ready to use. 2. EliA™ IgG Conjugate 50 or 200: β-Galactosidase labeled anti-IgG (mouse monoclonal antibodies) in PBS containing BSA and 0.06% sodium azide – six wedge shaped bottles, 5 mL each, ready to use; or six wedge-shaped bottles, 19 mL each, ready to use. 3. EliA™ Sample Diluent PBS containing BSA, detergent and 0.095% sodium azide – six bottles, 48 mL each, ready to use; or six bottles, 400 mL each, ready to use. 4. EliA™ Thyroid Positive Control 250 or 2500/5000 Human serum containing IgG antibodies to TG and TPO in PBS containing BSA, detergent and 0.095% sodium azide – six single use vials, 0.3 mL each, ready to use. 5. EliA™ Negative Control 250 or 2500/5000: Human sera from healthy donors in PBS containing BSA, detergent and 0.095% sodium azide – six single-use vials, 0.3 mL each, ready to use. 6. EliA™ IgG Calibrator Strips: Human IgG (0, 4, 10, 20, 100, 600 μg/L) in PBS containing BSA, detergent and 0.095% sodium azide – five strips, six single-use vials per strip, 0.3 mL each, ready to use. 7. EliA™ IgG Curve Control Strips: Human IgG (20 μg/L) in PBS containing BSA, detergent and 0.095% sodium azide – five strips, six single-use vials per strip, 0.3 mL each, ready to use. 3 {3} 8. EliA™ IgG Calibrator Well: Coated with mouse monoclonal antibodies – four carriers (12 wells each), ready to use. J. Substantial Equivalence Information: 1. Predicate device name(s) and 510(k) numbers: VarelisA TG Antibodies (K003414) VarelisA TPO Antibodies (K993585) Negative controls (K091845) Calibrators (K061165) 2. Comparison with predicate: | Similarities: EliA™ anti-TG Immunoassay | | | | --- | --- | --- | | Item | New Device | VarelisA TG Antibodies K003414 | | Intended Use | EliA anti-TG is intended for the in vitro quantitative measurement of IgG antibodies directed to thyroglobulin (TG) in human serum and plasma (heparin, EDTA) as an aid in the clinical diagnosis of autoimmune thyroiditis and Graves' disease. EliA anti-TG uses the EliA IgG method on the instruments Phadia 250 and Phadia 2500/5000. | The VarelisA TG (Thyroglobulin) Antibodies EIA kit is designed for the quantitative and qualitative determination of thyroglobulin antibodies in serum or plasma to aid in the diagnosis of thyroid diseases such as autoimmune thyroiditis and Graves' disease. | | Assay Type | ELISA | Same | | Type of test | Quantitative | Same | | Antigen | Human Thyroglobulin | Same | | Differences: EliA™ anti-TG Immunoassay | | | | --- | --- | --- | | Item | New Device | VarelisA TG Antibodies K003414 | | Instrument | Phadia 250 and 2500/5000, fully automated immunoassay analyzers | ELISA-Reader | | Instrumentation | Phadia 250 and 2500/5000 are fully Automated immunoassay analyzers | ELISA-Reader | | Reaction Temperature | 37°C (controlled) | Room temperature | | Assay Range | 12–4794 IU/mL | 3–3000 IU/mL | | Incubation times | Diluted patient samples: 30 minutes Conjugate: 28 minutes Development Solution: 39 minutes | Positive and negative controls, diluted patient samples: 30 minutes. Conjugate: 30 minutes Substrate: 10 minutes (in dark) | | Detection antibody (conjugate) | IgG conjugate: anti-human IgG β-Galactosidase (mouse monoclonal | IgG conjugate: anti-human IgG horse radish peroxidase (goat) | {4} | Differences: EliA™ anti-TG Immunoassay | | | | --- | --- | --- | | Item | New Device | VarelisA TG Antibodies K003414 | | | antibodies) | | | Signal | Fluorescence | Optical density (at 450nm) | | Calibrators | Six vials of human IgG at concentrations of 0, 4, 10, 20, 100 and 600 μg/L | Six vials of TG-specific IgG at concentrations of 0, 30, 100, 300, 1000 and 3000 IU/mL | | Cut-off | < 40 IU/mL negative 40–60 IU/mL equivocal > 60 IU/mL positive | < 60 IU/mL negative 60–100 IU/mL equivocal > 100 IU/mL positive | | Calibration curve | Option to store curve for up to 28 days and run curve controls in each assay for calibration | N/A | | Substrate | Development Solution 0.01 % 4-Methylumbelliferyl-β-D-galactoside and < 0.0010% preservative (mixture of 5-chloro-2-methyl-2H-isothiazol-3-one [EC no. 247-500-7] and 2-Methyl-2H-isothiazol-3-one [EC no. 220-239-6] at a 3:1 ratio. | TMB Chromogen | | Sample Dilution | 1:100 | 1:101 | | Concept | Modular reagents concept (test-method specific and general reagents) | All reagents in a single kit | | Similarities: EliA™ anti-TPO Immunoassay | | | | --- | --- | --- | | Item | New Device | VarelisA TG Antibodies K993585 | | Intended Use | EliA anti-TPO is intended for the in vitro quantitative measurement of IgG antibodies directed to thyroid peroxidase (TPO) in human serum and plasma (heparin, EDTA) as an aid in the clinical diagnosis of autoimmune thyroiditis and Graves' disease. EliA anti-TPO uses the EliA IgG method on the instruments Phadia 250 and Phadia 2500/5000. | The VarelisA TPO Antibodies EIA kit is designed for the quantitative and qualitative determination of TPO (thyroid peroxidase) antibodies in serum or plasma to aid in the diagnosis of thyroid diseases such as autoimmune thyroiditis and Graves' disease. | | Assay Type | ELISA | Same | | Type of teat | Quantitative | Same | | Antigen | Human Recombinant TPO | Same | {5} | Differences: EliA™ anti-TPO Immunoassay | | | | --- | --- | --- | | Item | New Device | VarelisA TG Antibodies K993585 | | Instrumentation | Phadia 250 and 2500/5000, fully automated immunoassay analyzers | ELISA-Reader | | Reaction Temperature | 37°C (controlled) | Room temperature | | Assay Range | 4–1542 IU/mL | 1–3000 IU/mL | | Incubation times | Diluted patient samples: 30 minutes Conjugate: 28 minutes Development Solution: 39 minutes | Positive and negative controls: diluted patient samples: 30 minutes Conjugate: 30 minutes Substrate: 10 minutes (in dark) | | Detection antibody (conjugate) | IgG conjugate: anti-human IgG β-Galactosidase (mouse monoclonal antibodies) | IgG conjugate: anti-human IgG horse radish peroxidase (goat) | | Signal | Fluorescence | Optical density (at 450nm) | | Calibrators | Six vials of human IgG at concentrations of 0, 4, 10, 20, 100 and 600 μg/L | Six vials of TG-specific IgG at concentrations of 0, 30, 100, 300, 1000 and 3000 IU/mL | | Cut-off | < 25 IU/mL negative 25–35 IU/mL equivocal > 35 IU/mL positive | < 60 IU/mL negative 60–100 IU/mL equivocal > 100 IU/mL positive | | Calibration curve | Option to store curve for up to 28 days and run curve controls in each assay for calibration | N/A | | Substrate | Development Solution 0.01 % 4-Methylumbelliferyl-β-D-galactoside and <0.0010% preservative (mixture of 5-chloro-2-methyl-2H-isothiazol-3-one [EC no. 247-500-7] and 2-Methyl-2H-isothiazol-3-one [EC no. 220-239-6] at a 3:1 ratio | TMB Chromogen | | Sample Dilution | 1:100 | 1:101 | | Concept | Modular reagents concept (test-method specific and general reagents) | All reagents in a single kit | # K. Standard/Guidance Document Referenced (if applicable): CLSI EP6-A - Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline CLSI EP17-A2 - Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures; Approved Guideline Second Edition. CLSI H18-A3: Procedures for the Handling and Processing of Blood Specimens for Common Laboratory Tests; Approved Guideline {6} L. Test Principle: The EliA™ test wells are coated with a human thyroglobulin protein, or a human recombinant thyroid peroxidase protein. If present in the patient's specimen, antibodies to TG or TPO bind to their specific antigen. After washing away unbound antibodies, enzyme-labeled antibodies against human IgG antibodies (EliA IgG Conjugate) are added to form an antibody-conjugate complex. After incubation, non-bound conjugate is washed away and the bound complex is incubated with a Development Solution. After stopping the reaction, the fluorescence in the reaction mixture is measured. The higher the value of fluorescent signal detected by the instrument, the higher the amount of antibody bound and detected in the sample tested. To evaluate test results, the response for patient samples is compared directly to the response for calibrators. M. Performance Characteristics (if/when applicable): 1. Analytical performance: Note: All results below met the manufacturer's pre-specified acceptance criteria. a. Precision/Reproducibility: The precision studies were performed using eight patient serum samples selected from different parts of the claimed measuring range of the assay on both Phadia 250 and Phadia 2500/5000 instruments to establish intra- and inter-assay precision. A total of 21 runs (three instruments/seven runs each) over seven days were performed on samples with 252 replicate determinations per sample with a calibration curve in each run. Two lots of $\mathrm{EliA}^{\mathrm{TM}}$ anti-TG well were included in the study. On each instrument the same eight samples were tested. All samples were neat specimens from different patients. The study was performed on three Phadia 250 and three Phadia 2500/5000 instruments. | EliA™ anti-TG on Phadia 250 (n = 252) | | | | | | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | Mean (IU/mL) | Inter-Run | | Intra-Run | | Inter-Instrument | | Total | | | | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | | Sample 1 | 30.6 | 1.5 | 4.9 | 2.7 | 8.7 | 2.0 | 6.4 | 3.6 | 11.9 | | Sample 2 | 37.3 | 1.8 | 4.7 | 2.3 | 6.2 | 3.1 | 8.4 | 4.3 | 11.4 | | Sample 3 | 74.9 | 0.7 | 0.9 | 3.2 | 4.3 | 3.7 | 5.0 | 5.0 | 6.7 | | Sample 4 | 124.9 | 0.9 | 0.7 | 4.5 | 3.6 | 3.5 | 2.8 | 5.8 | 4.6 | | Sample 5 | 184.8 | 1.1 | 0.6 | 5.2 | 2.8 | 0.8 | 0.5 | 5.7 | 3.1 | | Sample 6 | 771.5 | 8.8 | 1.1 | 21.2 | 2.7 | 0.8 | 0.1 | 24.8 | 3.2 | | Sample 7 | 2065.1 | 29.7 | 1.4 | 67.9 | 3.3 | 38.6 | 1.9 | 88.6 | 4.3 | | Sample 8 | 4147.2 | 54.5 | 1.3 | 164.6 | 4.0 | 85.6 | 2.1 | 194.6 | 4.7 | {7} | EliA™ anti-TG on Phadia 2500/5000 (n = 252) | | | | | | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | Mean (IU/mL) | Inter-Run | | Intra-Run | | Inter-Instrument | | Total | | | | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | | Sample 1 | 31.4 | 1.0 | 3.2 | 4.3 | 13.5 | 3.4 | 10.9 | 5.6 | 17.8 | | Sample 2 | 38.9 | 0.9 | 2.3 | 4.6 | 11.9 | 4.1 | 10.6 | 6.2 | 16.0 | | Sample 3 | 76.7 | 1.5 | 1.9 | 4.0 | 5.2 | 5.4 | 7.0 | 7.1 | 9.2 | | Sample 4 | 134.5 | 2.7 | 2.0 | 7.1 | 5.3 | 8.0 | 5.9 | 11.1 | 8.3 | | Sample 5 | 193.0 | 4.0 | 2.0 | 8.4 | 4.3 | 5.3 | 2.8 | 10.8 | 5.6 | | Sample 6 | 754.2 | 9.1 | 1.2 | 39.1 | 5.2 | 7.5 | 1.0 | 40.8 | 5.4 | | Sample 7 | 1869.6 | 0.0* | 0.0* | 100.5 | 5.4 | 79.5 | 4.3 | 128.4 | 6.9 | | Sample 8 | 3797.5 | 50.9 | 1.3 | 338.9 | 8.9 | 118.1 | 3.1 | 387.5 | 10.2 | * Inter-Run standard deviation of 0.0 and %CV of 0.0 indicates that this part of the total variation could not be determined as the Intra-Run variation completely explained the total variation. | EliA™ anti-TPO on Phadia 250 (n = 252) | | | | | | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | Mean (IU/mL) | Inter-Run | | Intra-Run | | Inter-Instrument | | Total | | | | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | | Sample 1 | 15.7 | 0.4 | 2.3 | 0.9 | 5.7 | 0.9 | 5.9 | 1.4 | 8.7 | | Sample 2 | 27.4 | 0.4 | 1.5 | 1.3 | 4.6 | 0.8 | 2.9 | 1.6 | 5.7 | | Sample 3 | 39.4 | 0.2 | 0.5 | 1.5 | 3.8 | 1.8 | 4.5 | 2.4 | 6.2 | | Sample 4 | 66.7 | 0.7 | 1.0 | 2.6 | 3.9 | 1.2 | 1.7 | 3.0 | 4.5 | | Sample 5 | 218.0 | 2.8 | 1.3 | 10.0 | 4.6 | 1.4 | 0.7 | 10.6 | 4.9 | | Sample 6 | 480.1 | 4.7 | 1.0 | 22.7 | 4.7 | 6.5 | 1.4 | 24.7 | 5.2 | | Sample 7 | 814.9 | 3.2 | 0.4 | 31.9 | 3.9 | 19.6 | 2.4 | 37.8 | 4.6 | | Sample 8 | 1212.6 | 0.0* | 0.0* | 48.0 | 4.0 | 57.6 | 4.8 | 75.0 | 6.2 | * Inter-Run standard deviation of 0.0 and %CV of 0.0% indicates that this part of the total variation could not be determined as the Intra-Run variation completely explained the total variation. {8} 9 | EliA™ anti-TPO on Phadia 2500/5000 (n = 252) | | | | | | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | Mean (IU/mL) | Inter-Run | | Intra-Run | | Inter-Instrument | | Total | | | | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | | Sample 1 | 17.7 | 0.4 | 2.2 | 1.5 | 8.7 | 1.6 | 8.9 | 2.3 | 13.1 | | Sample 2 | 26.6 | 0.8 | 3.1 | 1.7 | 6.4 | 0.4 | 1.5 | 2.1 | 8.0 | | Sample 3 | 38.5 | 1.3 | 3.4 | 2.3 | 5.9 | 0.9 | 2.3 | 2.8 | 7.3 | | Sample 4 | 65.3 | 1.7 | 2.6 | 4.0 | 6.1 | 1.1 | 1.7 | 4.5 | 7.0 | | Sample 5 | 202.9 | 5.4 | 2.7 | 14.0 | 6.9 | 3.4 | 1.7 | 15.4 | 7.6 | | Sample 6 | 466.6 | 10.2 | 2.2 | 33.7 | 7.2 | 7.2 | 1.5 | 36.5 | 7.8 | | Sample 7 | 802.8 | 13.8 | 1.7 | 59.1 | 7.4 | 16.3 | 2.0 | 62.8 | 7.8 | | Sample 8 | 1340.2 | 28.0 | 2.1 | 120.2 | 9.0 | 30.8 | 2.3 | 127.5 | 9.5 | b. Linearity/assay reportable range: For EliA™ anti-TG, seven patient serum samples and for EliA™ anti-TPO, six patient samples were serially diluted using EliA Sample Diluent and tested in three replicates with one batch of EliA™ anti-TG immunoassay and EliA™ anti-TPO immunoassay and one set of system reagents on the Phadia® 250 or Phadia® 2500/5000 instruments. The observed values were graphed against the calculated values and a linear regression was performed. Results are summarized below: | EliA™ anti-TG on Phadia 250 | | | | | | --- | --- | --- | --- | --- | | Sample | Dilution Range (IU/mL) | Slope (95% CI) | Intercept (95% CI) | R² | | 1 | 2655.8–31.1 | 1.002 (0.979–1.024) | -1.46 (-24.59–21.66) | 0.9990 | | 2 | 2935.0–33.5 | 1.008 (0.979–1.037) | 13.21 (-19.92–46.35) | 0.9983 | | 3 | 4827.9–62.5 | 1.025 (0.983–1.006) | 58.32 (-19.33–135.98) | 0.9967 | | 4 | 375.3–10.1 | 0.998 (0.991–1.005) | 1.01 (-0.07–2.09) | 0.9999 | | 5 | 468.8–17.5 | 0.995 (0.976–1.015) | 1.34 (-2.73–5.42) | 0.9995 | {9} | EliATM anti-TG on Phadia 2500/5000 | | | | | | --- | --- | --- | --- | --- | | Sample | Dilution Range (IU/mL) | Slope (95% CI) | Intercept (95% CI) | R2 | | 1 | 2626.4–31.1 | 0.995 (0.977–1.04) | 23.48 (4.45–42.51) | 0.9993 | | 2 | 2052.8–36.2 | 1.008 (0.986–1.031) | 17.61 (0.89–36.12) | 0.9991 | | 3 | 5127.8–63.2 | 1.006 (0.974–1.037) | 60.01 (-1.87–121.88) | 0.9981 | | 4 | 375.9–11.1 | 0.977 (0.928–1.026) | 8.21 (0.42–16.01) | 0.9962 | | 5 | 470.7–12.1 | 1.01 (0.987–1.034) | 2.09 (-2.60–6.78) | 0.9992 | | 6 | 412.0–15.4 | 0.987 (0.959–1.015) | -1.38 (-6.53–3.78) | 0.9990 | | 7 | 327.9–13.6 | 1.016 (0.969–1.063) | 4.09 (-2.78–10.96) | 0.9973 | The technical measuring range (detection limit, upper limit of the calibration curve) for $\mathrm{EliA}^{\mathrm{TM}}$ anti-TG is from 12 to $4797\mathrm{IU / mL}$ . The upper limit of the reported results in $\mathrm{EliA IU / ml}$ can vary due to a lot-specific conversion from $\mu \mathrm{g} / \mathrm{L}$ to $\mathrm{EliA IU / mL}$ . The transfer of the upper limit of the measuring range to $\mathrm{IU / ml}$ is defined by the formula: $600~\mu \mathrm{g} / 1\times 9.98$ (for anti-TG), where $600~\mu \mathrm{g} / \mathrm{L}$ is the highest calibration point. Results above the upper limit are generally reported as "above". Linearity was shown for the range between $10.1 - 4827.9\mathrm{IU / mL}$ on Phadia 250, and between $11.1 - 5127.8$ on Phadia 2500/5000. The claimed linear range of $\mathrm{EliA}^{\mathrm{TM}}$ anti-TG for instruments is 12-4797 $\mathrm{IU / mL}$ . The labeling states that due to differing binding characteristics of the antibodies in patient samples, not all sera can be diluted linearly within the technical measuring range. {10} | EliATM anti-TPO on Phadia 250 | | | | | | --- | --- | --- | --- | --- | | Sample | Dilution Range (IU/mL) | Slope (95% CI) | Intercept (95% CI) | R2 | | 1 | 1536.1–16.5 | 1.004 (0.996–1.013) | -1.78 (-6.68–3.12) | 0.9999 | | 2 | 2201.2–22.1 | 1.048 (0.993–1.102) | -1.48 (-47.76–44.80) | 0.9946 | | 3 | 479.8–7.0 | 1.008 (0.999–1.017) | -0.50 (-2.28–1.28) | 0.9999 | | 4 | 310.9–3.2 | 1.005 (0.996–1.015) | 0.42 (-0.71–1.55) | 0.9998 | | 5 | 349.2–6.3 | 0.993 (0.983–1.003) | 0.90 (-0.47–2.27) | 0.9998 | | 6 | 458.0–5.6 | 0.998 (0.988–1.007) | 1.33 (-0.34–3.01) | 0.9998 | | EliATM anti-TPO on Phadia 2500/5000 | | | | | | --- | --- | --- | --- | --- | | Sample | Dilution Range (IU/mL) | Slope (95% CI) | Intercept (95% CI) | R2 | | 1 | 1400.8–15.1 | 0.987 (0.968–1.005) | -0.70 (-10.60–9.20) | 0.9993 | | 2 | 1955.9–22.1 | 1.008 (0.996–1.020) | 2.43 (-6.55–11.42) | 0.9997 | | 3 | 471.6–4.6 | 0.988 (0.970–1.007) | 1.55 (-1.76–4.85) | 0.9993 | | 4 | 205.7–3.6 | 1.071 (1.054–1.087) | 0.40 (-0.56–1.36) | 0.9996 | | 5 | 349.4–5.8 | 0.997 (0.968–1.005) | 0.90 (-0.32–2.12) | 0.9999 | {11} | 6 | 464.3–5.3 | 0.987 (0.970–1.004) | 1.61 (-1.46–4.68) | 0.9994 | | --- | --- | --- | --- | --- | Due to an administrative error, the reference to “anti-TG” in the paragraph below was erroneously included, and has been replaced with the correct reference to “anti-TPO”. The technical measuring range (LoD/LoQ, upper limit of the calibration curve) for EliA™ anti-TPO is from 4 to 1542 IU/mL. The upper limit of the reported results in EliA IU/mL can vary due to a lot-specific conversion from µg/L to EliA IU/mL. The conversion of the upper limit of the measuring range to IU/mL is calculated by the formula: 600 µg/L x 3.21 (for anti-TPO anti-TG), where 600 µg/L is the highest calibration point. Results above the upper limit are generally reported as “above”. Linearity was shown for the range between 3.2–2201.2 U/mL on Phadia 250, and 3.6–1955.9 on Phadia 2500/5000. The claimed linear range of EliA™ anti-TPO for instruments is 4–1542 IU/mL. The labeling states that due to differing binding characteristics of the antibodies in patient samples, not all sera can be diluted linearly within the technical measuring range. ## High dose Hook effect: Hook effect was investigated by using five serum samples above the upper limit of the measuring range. Five high positive samples were diluted and the dilutions were measured in two replicates and compared to the upper limit of the technical measuring range. EliA™ anti-TG Immunoassay: No hook effect was observed when analyzing a high positive serum sample up to 135,582 IU/mL. EliA™ anti-TPO Immunoassay: No hook effect was observed when analyzing a high positive serum sample up to 20,725 IU/mL. Results above the upper limit of the measuring range are reported as “above”. Package insert recommends not making dilution of samples outside measuring range. c. Traceability, Stability, Expected values (controls, calibrators, or methods): i) Traceability Calibrators – The IgG calibrators are traceable (via unbroken chain of calibrations) to the International Reference Preparation (IRP) 67/86 of Human Serum Immunoglobulins A, G and M from the World Health Organization (WHO). New batches of IgG calibrators are compared to a secondary standard (standardized with the IRP) or the IRP directly and adjusted accordingly to meet the correct concentration. EliA™ anti-TG is calibrated against the 1st International Reference Preparation for human anti-TG serum code 65/93 and EliA™ anti-TPO is calibrated against the NIBSC research standard for anti-thyroid microsome serum code 66/387. The {12} instrument measures specific IgG concentrations in $\mu \mathrm{g} / \mathrm{L}$ . By using a conversion factor given by the lot-specific code of the EliA™ test well, the results are automatically converted to IU/mL for EliA™ anti-TG and EliA™ anti-TPO. The calibrators are a set of six WHO-standardized IgG calibrators derived from human serum with assigned values from $0 - 600\mu \mathrm{g / mL}$ . The calibrator curve is acquired by fitting the values of the six calibrators and can be stored for up to 28 days by the instrument to be used on additional assays. Each additional assay includes calibrator (curve) controls that have to recover in defined ranges to ensure that the stored calibration curve is still valid. Controls - EliA Thyroid Positive Control is prepared from selected pooled human sera and contains IgG antibodies to TG and TPO. EliA IgG/IgM/IgA Negative Control is prepared from selected pooled sera from normal, healthy donors. The controls are pre-diluted and ready for use. Each EliA™ Control package contains a Control Certificate listing predefined acceptance criteria for the EliA™ products the Controls can be used with. The target ranges of the EliA™ Controls tested with the EliA™ TG and TPO on the two Phadia® instruments are summarized below: | Instrument | EliATM Positive Control-TG | EliATM Positive Control-TPO | | --- | --- | --- | | Phadia® 250 | 467.9–1091.7 IU/mL | 104.1–243.0 IU/mL | | Phadia® 2500/5000 | 505.3–1179.1 IU/mL | 100.0–233.3 IU/mL | | Instrument | EliATM IgG/IgM/IgA Negative Control | EliATM IgG/IgM/IgA Negative Control | | Phadia® 250 | < 40 IU/mL | < 25 IU/mL | | Phadia® 2500/5000 | < 40 IU/mL | < 25 IU/mL | # ii) Kit Stability: Data for open and closed real time stability and on-board stability of EliA™ IgG reagents and general EliA reagents on Phadia 250 and Phadia 2500/5000 instrument were submitted and cleared with several other EliA tests, e.g., K141375 for EliA M2, for Phadia 250 instrument and with K061165 for EliA CCP for the Phadia 2500/5000. Shelf-life stability - The real time stability test confirmed the shelf life for EliA™ anti-TG wells and EliA™ anti-TPO wells and it was determined to be 18 months. The shelf life stability of the EliA™ Thyroid Positive Control was determined to be 15 months. The shelf life stability of the EliA IgG/IgM/IgA Negative Control was 24 months. Open Stability - Stability of the foil bag containing the EliA anti-TG and EliA anti-TPO wells after first opening was tested and determined to be 9 months at $2 - 8^{\circ}\mathrm{C}$ . {13} Controls - EliA™ Thyroid Positive Control 250, EliA™ Thyroid Positive Control 2500/5000, EliA™ IgG/IgM/IgA Negative Control 250, and EliA™ IgG/IgM/IgA Negative Control 2500/5000 are for single use only. Therefore, stability after first opening study is not required. On-board stability – The on-board stability EliA™ anti-TG and EliA™ anti-TPO carriers (containing the antigen coated wells) were tested over four weeks using three positive and two negative samples only on the Phadia 250 instrument. As the storage conditions in the Phadia 2500/5000 instrument are similar, the results can also be used for stability claims for the Phadia 2500/5000. The on-board stability for the Phadia 250 instrument was determined to be 28 days at $2 - 8^{\circ}\mathrm{C}$ . # iii) Sample Storage The package insert recommends following the guidelines in CLSI H18-A3: Procedures for the Handling and Processing of Blood Specimens for Common Laboratory Tests; Approved Guideline for sample storage. Separated serum/plasma should remain at room temperature for no longer than 8 hours. If assays will not be completed within 8 hours, serum/plasma should be refrigerated $(2 - 8^{\circ}\mathrm{C})$ . If assays are not completed within 48 hours, or the separated serum/plasma will be stored beyond 48 hours, serum/plasma should be frozen at or below $-20^{\circ}\mathrm{C}$ . Freezing and thawing should be avoided. # d. Detection limit: The Limit of Blank (LoB) and Limit of Detection (LoD) were determined in accordance with CLSI EP17-A2. On each Phadia instrument (Phadia® 250 and Phadia® 2500/5000), four immunoglobulin depleted serum (for LoB) and four low antibody samples (for LoD) were assayed in six replicates in each of six runs at six different days (36 replicates per sample). For both instrument types (Phadia 250 and Phadia 2500/5000), each sample was run in a total of six runs on two instruments (3 runs on instrument #1 and three runs on instrument #2) with two lots of wells and reagents. The results are summarized in the tables below: | EliA™ anti-TG | LoB (IU/mL) | LoD (IU/mL) | | --- | --- | --- | | Phadia 250 | 5.0 | 10.1 | | Phadia 2500/5000 | 4.9 | 8.5 | | A single LoD of 12.0 IU/mL for anti-TG was chosen. | | | | EliA™ anti-TPO | LoB (IU/mL) | LoD (IU/mL) | | Phadia 250 | 1.7 | 2.9 | | Phadia 2500/5000 | 1.5 | 2.8 | | A single LoD of 4 IU/mL for anti-TPO was chosen. | | | {14} Limit of Quantitation (LoQ) For LoQ determination for both assays, bias was set to $12\%$ which leads to estimated $\mathrm{CV}_{\mathrm{tot}}$ of $30\%$ . The target value for the total error $= 12 + 2^{*}\% \mathrm{CV} = 72\%$ . Because estimated total error for the LoD in all cases was less than the target value, the LoQ was determined to equal the LoD for both assays. # e. Analytical specificity: # i) Endogenous Interference Interferences were assessed by testing three samples (one negative, one around the cut-off, and a high positive sample). Each sample was spiked with the interfering substances or substance-specific blanks, and analyzed in two runs, each in duplicates $(n = 4)$ , on one lot of $\mathrm{EliA}^{\mathrm{TM}}$ TG or $\mathrm{EliA}^{\mathrm{TM}}$ TPO Well and one lot of each system reagents. The data demonstrated that $\mathrm{EliA}^{\mathrm{TM}}$ TG or $\mathrm{EliA}^{\mathrm{TM}}$ TPO was not adversely affected (percent recoveries ranged from 94 to $109\%$ for $\mathrm{EliA}^{\mathrm{TM}}$ TG and 93 to $105\%$ for $\mathrm{EliA}^{\mathrm{TM}}$ TPO) by high levels of the following substances up to the concentrations listed in the tables below: | EliATM anti-TG and EliATM anti-TPO Immunoassay | | | --- | --- | | Potential Interfering Compound | Concentration in undiluted sample | | Bilirubin F | 192 mg/dL | | Bilirubin C | 201 mg/dL | | Hemoglobin | 4960 mg/dL | | Lipemic factor | 1% | | Rheumatoid factor | 500 IU/mL | | Thyroxine | 2.5 ng/dL | | Iodide | 3 mM | # ii) Carry-over In order to demonstrate the efficacy of the Phadia 250 system of washing the sample pipette between samples, carry-over study was previously evaluated in K082759, EliA Ro assay. The data demonstrate no carryover effect as the results of the Diluent sample results were not affected by the results of the previous sample. Phadia 2500/5000 instruments use disposable tips for pipetting samples and a separate pipette for the conjugate, therefore carry-over from samples to conjugate is unlikely. {15} # f. Assay cut-off: The study was done on 604 apparently healthy blood donor samples from Caucasian, African American, Hispanic and Asian individuals almost equally distributed by sex and age, the expected values for each of the antigens in the submission in the normal population and to confirm the defined cut-off. Based on the results of the expected values/reference range study, the $90^{\text{th}}$ and $95^{\text{th}}$ percentile of the 604 samples were calculated and the $90^{\text{th}}$ percentile was taken into account for setting the cut-off. The assay cut-offs were set as follows: | EliATM anti-TG | | | --- | --- | | < 40 IU/mL | Negative | | 40–60 IU/mL | Equivocal | | > 60 IU/mL | Positive | In case of equivocal results, it is recommended to retest the patient after 8-12 weeks. | EliATM anti-TPO | | | --- | --- | | < 25 IU/mL | Negative | | 25–35 IU/mL | Equivocal | | > 35 IU/mL | Positive | In case of equivocal results, it is recommended to retest the patient after 8-12 weeks. # 2. Comparison studies: # a. Method comparison with predicate device: EliATM anti-TG Immunoassay: A total of 718 clinically defined serum samples from patients with autoimmune thyroiditis or Graves' disease, non-autoimmune thyroid diseases, bacterial or viral infections, cancer or connective tissue diseases, type II diabetes mellitus, Sjögren's syndrome, rheumatoid arthritis, hypergammaglobulinemia, Systemic lupus erythematosus (SLE), ulcerative colitis, celiac disease, Crohn's disease, type I diabetes, chronic thyroiditis, sub-acute thyroiditis, multi-nodular goiter, pernicious anemia, myasthenia gravis, miscarriage, pre-eclampsia, thyroid carcinoma and pregnant women across all trimesters (see table below under clinical studies) were tested on EliATM anti-TG (new device) and VarelisA TG Antibodies (predicate device) using Phadia® 250 instrument. The results are summarized below: {16} | n=718 | VarelisA TG Abs. positive: > 100 IU/mL | VarelisA TG Abs. equivocal: 60–100 IU/mL | VarelisA TG Abs. negative: < 60 IU/mL | Total | | --- | --- | --- | --- | --- | | EliATM anti-TG positive: > 60 IU/mL | 142 | 25 | 9 | 176 | | EliATM anti-TG Equiv.: 40–60 IU/mL | 10 | 13 | 13 | 36 | | EliATM anti-TG negative: < 40 IU/mL | 7 | 28 | 471 | 506 | | Total | 159 | 66 | 493 | 718 | | Positive percent agreement: 89.3% (142/159) | | 95% CI: 83.1–93.6% | | | | Equivocal percent agreement: 19.7% (13/66) | | 95% CI: 10.9–31.3% | | | | Negative percent agreement: 95.5% (471/493) | | 95% CI: 93.3–97.2% | | | | Total percent agreement: 87.2% (616/718) | | 95% CI: 84.5–89.5% | | | For the calculation of agreement, from 718 samples, 280 samples (260 samples below and 20 samples above the measuring range limits) were excluded resulting in a total of 438 samples. Agreements were calculated by grouping EliATM TG equivocal results with its test negative results, and then agreements were calculated again by grouping EliATM TG equivocal results with the test positive results: | Equivocal EliATM TG results considered as negative | | | | | | --- | --- | --- | --- | --- | | n = 438 | VarelisA TG (IU/mL) | | Total | | | | | Positive: > 100 | | Negative: ≤ 100 | | EliATM TG (IU/mL) | Positive: > 60 | 122 | 34 | 156 | | | Negative: ≤ 60 | 14 | 268 | 282 | | Total | | 136 | 302 | 438 | | Positive percent agreement: 89.7% (122/136) | | 95% CI: 83.3–94.3% | | | | Negative percent agreement: 88.7% (269/302) | | 95% CI: 84.6–92.1% | | | | Total percent agreement: 89.0% (390/438) | | 95% CI: 85.7–91.8% | | | {17} 18 | Equivocal EliA™ TG results considered as positive | | | | | | --- | --- | --- | --- | --- | | n = 438 | VarelisA TG (IU/mL) | | Total | | | | | Positive: > 100 | | Negative: ≤ 100 | | EliA™ TG (IU/mL) | Positive: > 40 | 170 | 21 | 191 | | | Negative: ≤ 40 | 27 | 220 | 247 | | Total | | 197 | 241 | 438 | | Positive percent agreement: 86.3% (170/197) 95% CI: 80.7–90.8% | | | | | | Negative percent agreement: 91.3% (220/241) 95% CI: 87.0–94.5% | | | | | | Total percent agreement: 89.0% (390/438) 95% CI: 85.7–91.8% | | | | | EliA™ anti-TPO Immunoassay: A total of 718 clinically defined serum samples with a diagnosis patients with Hashimoto's or Graves' disease, non-autoimmune thyroid diseases, bacterial or viral infections, cancer or connective tissue diseases, type II diabetes mellitus, Sjögren's syndrome, rheumatoid arthritis, hypergammaglobulinemia, SLE, ulcerative colitis, celiac disease, Crohn's disease, type I diabetes, chronic thyroiditis, sub-acute thyroiditis, multi-nodular goiter, pernicious anemia, myasthenia gravis, miscarriage, pre-eclampsia, thyroid carcinoma and pregnant women across all trimesters (refer to the Table 3 in Clinical studies) were tested on EliATM anti-TPO (new device) and VarelisA TPO Antibodies (predicate device) using Phadia® 250 instrument. Additionally, 77 technical samples without clinical diagnosis were also included in the test on EliA anti-TPO (new device) and VarelisA TPO Antibodies (predicate device) to cover the equivocal range. The technical samples are blood samples without a clinical diagnosis. So, the total number of samples tested was 795 (718 + 77). The results are summarized below: | n=795 | VarelisA TPO Abs. positive: > 100 (IU/mL) | VarelisA TPO Abs. equivocal: 60–100 (IU/mL) | VarelisA TPO Abs. negative: < 60 (IU/mL) | Total | | --- | --- | --- | --- | --- | | EliA™ TPO positive: > 35 (IU/mL) | 191 | 24 | 10 | 225 | | EliA™ TPO Equiv.: 25–35 (IU/mL) | 0 | 22 | 65 | 87 | | EliA™ TPO negative: < 25 (IU/mL) | 0 | 0 | 483 | 483 | | Total | 191 | 46 | 558 | 795 | | Positive percent agreement: 100% (191/191) 95% CI: 98.1–100% | | | | | | Equivocal percent agreement: 47.8% (22/46) 95% CI: 32.9–63.1% | | | | | | Negative percent agreement: 86.6% (483/558) 95% CI: 83.4–89.3% | | | | | | Total percent agreement: 87.5% (696/795) 95% CI: 85.0–89.8% | | | | | {18} For the calculation of agreement, 297 samples (273 samples below and 24 samples above the measuring range limits) were excluded resulting in a total of 498 samples. Agreements were calculated by grouping $\mathrm{EliA}^{\mathrm{TM}}$ TPO equivocal results with the test negative results, and then agreements were calculated again by grouping $\mathrm{EliA}^{\mathrm{TM}}$ TPO equivocal results with the test positive results: | Equivocal EliATM anti-TPO results considered as negative | | | | | | --- | --- | --- | --- | --- | | n = 498 | VarelisA TPO (IU/mL) | | Total | | | | | Positive: >100 | | Negative: ≤100 | | EliATM TPO (IU/mL) | Positive: >35 | 167 | 34 | 201 | | | Negative: ≤35 | 0 | 297 | 297 | | Total | | 167 | 331 | 498 | | Positive percent agreement: 100% (167/167) 95% CI: 97.8–100% Negative percent agreement: 89.7.6% (297/331) 95% CI: 85.9–92.8% Total percent agreement: 93.2% (464/498) 95% CI: 90.6–95.2% | | | | | | Equivocal EliATM anti-TPO results considered as positive | | | | | | --- | --- | --- | --- | --- | | n = 498 | VarelisA TPO (IU/mL) | | Total | | | | | Positive: >60 | | Negative: ≤60 | | EliATM TPO (IU/mL) | Positive: >25 | 213 | 75 | 288 | | | Negative: ≤25 | 0 | 210 | 210 | | Total | | 213 | 285 | 498 | | Positive percent agreement: 100% (213/213) 95% CI: 98.3–100% Negative percent agreement: 73.7% (210/285) 95% CI: 68.2–78.7% Total percent agreement: 84.9% (423/498) 95% CI: 81.5–88.0% | | | | | # b. Matrix comparison: The purpose of this study was to demonstrate that the new device $\mathrm{EliA}^{\mathrm{TM}}$ anti-TG and $\mathrm{EliA}^{\mathrm{TM}}$ anti-TPO give the same results for serum, Li-heparin plasma and K3-EDTA plasma collected from the same patient. A total of 57 matrix-matched samples spread across the assay range were assayed in duplicate on the Phadia 250. Out of 57 samples, 25 samples were spiked with serum samples of high antibody titer to cover the measuring range as it was not feasible to obtain a relevant number of matched serum- and plasma-samples from thyroid disease patients. However, the seven samples within the equivocal range and 25 negative samples were neat samples. Negative samples did not switch to positive in any serum/plasma combination. Weighted Deming regression plots were generated and the corresponding slopes of regression and coefficients were determined. The manufacturer's acceptance criterion was that the quotient of plasma to serum concentration should be between 0.80-1.20 for {19} all equivocal/positive sera. All of the samples met the acceptance criterion. The results are summarized in the tables below: | | EliA™ anti-TG Immunoassay | | | | | --- | --- | --- | --- | --- | | | Range Tested (IU/mL) | Slope (95% CI) | Intercept (95% CI) | R² | | Serum vs K3-EDTA Plasma | 25.5–4323.7 | 1.00 (0.97–1.03) | -3.16 (-6.79–0.47) | 1.00 | | Serum vs Li-Heparin Plasma | 24.3–5639.9 | 100 (0.97–1.03) | -0.43 (-3.96–3.11) | 1.00 | | | EliA™ anti-TPO Immunoassay | | | | | | Range Tested (IU/mL) | Slope (95% CI) | Intercept (95% CI) | R² | | Serum vs K3-EDTA Plasma | 10.8–1445.2 | 1.00 (0.90–1.10) | -3.07 (-8.90–2.75) | 0.99 | | Serum vs Li-Heparin Plasma | 11.0–1389.9 | 0.99 (0.95–1.03) | -0.78 (-2.25–0.68) | 0.99 | # c. Instrument comparison A study was performed to demonstrate that the performance of EliA™ anti-TG or EliA™ anti-TPO was equivalent on the Phadia® 250 and Phadia® 2500/5000 instruments. A total of 50 samples (30 positive, 8 equivocal and 12 negative) spanning the assay range were analyzed in six runs in single replicates on three Phadia®250 and three Phadia® 2500/5000 instruments, with two runs on each instrument. The results of a Passing Bablock regression analysis are shown below: | | EliA™ anti-TG Immunoassay | | | --- | --- | --- | | | Slope (95% CI) | Intercept (95% CI) | | Phadia®250 vs Phadia®2500/5000 | 0.96 (0.93–0.97) | 1.70 (0.70–3.80) | | | EliA™ anti-TPO Immunoassay | | | --- | --- | --- | | | Slope (95% CI) | Intercept (95% CI) | | Phadia®250 vs Phadia®2500/5000 | 0.98 (0.97–0.99) | 0.20 (-0.40–0.80) | {20} # 3. Clinical studies: a. Clinical sensitivity and specificity: # EliA™ anti-TG Immunoassay The performance of EliA™ anti-TG was compared to a clinical diagnosis autoimmune Thyroiditis. The validation set consisted of clinically characterized sera from AI Thyroiditis and various disease controls for a total of 718 patient samples. The results of EliA™ anti-TG for each disease category are shown below: | Condition | Number of samples | (%) positive on EliA™ TG | (%) positive on predicate | | --- | --- | --- | --- | | Graves` Disease | 100 | 48 (48%) | 48 (48%) | | Autoimmune thyroiditis | 115 | 72 (62.6%) | 69 (60%) | | Non-AI Thyroid Disease | 48 | 7 (14.6%) | 9 (18.8%) | | Connective Tissue Disease | 15 | 2 (13.3%) | 2 (13.3%) | | Crohn`s Disease | 20 | 1 (5%) | 0 (0%) | | Ulcerative Colitis | 20 | 1 (5%) | 0 (0%) | | Primary Biliary Cirrhosis | 10 | 0 (0%) | 0 (0%) | | HIV infection | 20 | 0 (0%) | 3 (15%) | | HCV infection | 20 | 1 (5%) | 2 (10%) | | HBV infection | 20 | 1 (5%) | 2 (10%) | | Other infection | 20 | 1 (5%) | 1 (5%) | | Cancer (Various) | 56 | 13 (23.2%) | 7 (12.5%) | | Rheumatoid Arthritis | 12 | 2 (16.7%) | 1 (8.3%) | | Hypergammaglobulinemia | 15 | 1 (6.7%) | 1 (6.7%) | | Systemic Lupus Erythematosus | 24 | 5 (20.8%) | 3 (12.5%) | | Sjögren`s Syndrome | 14 | 2 (14.3%) | 2 (14.3%) | | Celiac Disease | 15 | 3 (20%) | 2 (13.3%) | | Type I Diabetes Mellitus | 15 | 2 (13.3%) | 2 (13.3%) | | Type II Diabetes Mellitus | 15 | 1 (6.7%) | 1 (6.7%) | | Pregnant Women (all trimesters) | 90 | 5 (5.6%) | 2 (2.2%) | | Pre-eclampsia | 5 | 0 (0%) | 0 (0%) | | Miscarriage | 9 | 6 (66.7%) | 1 (11.1%) | | Thyroid Cancer | 21 | 0 (0%) | 0 (0%) | | Myasthenia gravis | 7 | 0 (0%) | 0 (0%) | | Pernicious Anemia | 8 | 2 (25%) | 2 (25%) | | Chronic Lymph. Thyroiditis | 10 | 0 (0%) | 0 (0%) | | Sub-acute Thyroiditis and Chron. Lymph. Thyroiditis | 5 | 5 (100%) | 5 (100%) | | Multi-nodular Goiter | 4 | 0 (0%) | 0 (0%) | {21} Clinical performance with equivocal samples considered negative is summarized in the following table: | | Diagnosis | | Total | | | --- | --- | --- | --- | --- | | | | Positive | | Negative | | EliATM anti-TG(IU/mL) | Positive: > 60 | 120 | 56 | 176 | | | Negative: ≤ 60 | 95 | 447 | 542 | | Total | | 215 | 503 | 718 | | Clinical sensitivity: | 55.8% (120/215) | 95% CI: 48.9–62.6% | | | | Clinical specificity: | 88.9% (447/503) | 95% CI: 85.8–91.5% | | | Clinical performance with equivocal samples considered positive is summarized in the following table: | | Diagnosis | | Total | | | --- | --- | --- | --- | --- | | | | Positive | | Negative | | EliATM anti-TG(IU/mL) | Positive: > 40 | 134 | 78 | 212 | | | Negative: ≤ 40 | 81 | 425 | 506 | | Total | | 215 | 503 | 718 | | Clinical sensitivity: | 62.3% (134/215) | 95% CI: 55.5–68.8% | | | | Clinical specificity: | 84.5% (425/503) | 95% CI: 81.0–87.5% | | | # EliATM anti-TPO Immunoassay The performance of EliATM anti-TPO was compared to a clinical diagnosis autoimmune Thyroiditis. The validation set consisted of clinically characterized sera from AI Thyroiditis and various disease controls for a total of 718 patient samples. The results of EliATM anti-TPO for each disease category are shown below: | Condition | Number of samples | (%) positive on EliATM TPO | (%) positive on predicate | | --- | --- | --- | --- | | Graves` Disease | 100 | 81 (81%) | 68 (68%) | | Autoimmune thyroiditis | 115 | 96 (83.5%) | 88 (76.5%) | | Non-AI Thyroid Disease | 48 | 5 (10.4%) | 5 (10.4%) | | Connective Tissue Disease | 15 | 1 (6.7%) | 1 (6.7%) | | Crohn`s Disease | 20 | 0 (0%) | 0 (0%) | | Ulcerative Colitis | 20 | 0 (0%) | 0 (0%) | | Primary Biliary Cirrhosis | 10 | 0 (0%) | 0 (0%) | | HIV infection | 20 | 1 (5%) | 0 (0%) | | HCV infection | 20 | 1 (5%) | 1 (5%) | | HBV infection | 20 | 0 (0%) | 0 (0%) | | Other infection | 20 | 0 (0%) | 0 (0%) | {22} | Cancer | 56 | 11 (20%) | 5 (9%) | | --- | --- | --- | --- | | Rheumatoid Arthritis | 12 | 1 (8%) | 1 (8%) | | Hypergammaglobulinemia | 15 | 1 (6%) | 0 (0%) | | Systemic Lupus Erythematosus | 24 | 1 (4%) | 0 (0%) | | Sjögren's Syndrome | 14 | 1 (7%) | 1 (7%) | | Celiac Disease | 15 | 2 (13%) | 2 (13%) | | Type I Diabetes Mellitus | 15 | 2 (13%) | 2 (13%) | | Type II Diabetes Mellitus | 15 | 2 (13%) | 2 (13%) | | Pregnant Women (all trimesters) | 90 | 10 (11%) | 10 (11%) | | Pre-eclampsia | 5 | 4 (80%) | 3 (60%) | | Miscarriage | 9 | 2 (22%) | 1 (11%) | | Thyroid Cancer | 21 | 2 (9%) | 1 (5%) | | Myasthenia gravis | 7 | 1 (14%) | 0 (0%) | | Pernicious Anemia | 8 | 1 (12%) | 0 (0%) | | Chronic Lymph. Thyroiditis | 10 | 1 (10%) | 1 (10%) | | Sub-acute Thyroiditis and Chron. | 5 | 4 (80%) | 4 (80%) | | Multi-nodular Goiter | 4 | 0 (0%) | 0 (0%) | Clinical performance with equivocal samples considered negative is summarized in the following table: | | Diagnosis | | Total | | | --- | --- | --- | --- | --- | | | | Positive | | Negative | | EliATM anti-TPO(IU/mL) | Positive: > 35 | 177 | 48 | 225 | | | Negative: ≤ 35 | 38 | 455 | 493 | | Total | | 215 | 503 | 718 | | Clinical sensitivity: | | 82.3% (177/215) | 95% CI: 76.6%-87.2% | | | Clinical specificity: | | 90.5% (455/503) | 95% CI: 87.5%-92.9% | | Clinical performance with equivocal samples considered positive is summarized in the following table: | | Diagnosis | | Total | | | --- | --- | --- | --- | --- | | | | Positive | | Negative | | EliATM anti-TPO(IU/mL) | Positive: > 25 | 182 | 53 | 235 | | | Negative: ≤ 25 | 33 | 450 | 483 | | Total | | 215 | 503 | 718 | | Clinical sensitivity: | | 84.7% (182/215) | 95% CI: 79.1%-89.2% | | | Clinical specificity: | | 89.5% (450/503) | 95% CI: 86.4%-92.0% | | b. Other clinical supportive data (when a. and b. are not applicable): Not applicable {23} 4. Clinical cut-off: Same as assay cut-off 5. Expected values/Reference range: A total of 604 apparently healthy, age- and gender-matched blood donor samples from a population to better reflect the ethnic composition of the US population (~60.5% White Americans, ~14% African Americans, ~17% Hispanics, ~6% Asians, ~2.5% others) were used. Note: The 90th percentile was chosen to determine the cut-off because according to literature &gt;10% of the healthy subjects are reported to have anti-thyroglobulin / anti-thyroid peroxidase antibodies. The results are summarized below: | | EliA™ TG IU/mL | | --- | --- | | Mean | 61.2 | | Median | 12.2 | | Range | 12–5071.7 | | 90th percentile | 35.6 | | 95th percentile | 97.9 | | | EliA™ TPO | | Mean | 17.3 | | Median | 4.4 | | Range | 4 – 1004.3 | | 90th percentile | 14.2 | | 95th percentile | 59.9 | EliA™ anti-TG Immunoassay: For EliA™ anti-TG, the 90th percentile lies at 35.6 IU/mL. The lower limit of the equivocal range was chosen to be at least equal to lowest positive calibrator (40 IU/mL). The upper cut-off value was determined as 56 IU/mL (40 IU/ml x 1.4) which was rounded up to 60 IU/mL. EliA™ anti-TPO Immunoassay: For EliA™ anti-TPO, the 90th percentile lies at 14.2 IU/ml. The lower limit of the equivocal range was chosen to be at least equal to lowest positive calibrator (25 IU/mL). The upper cut-off value was calculated as 35 IU/mL (25 IU/ml x 1.4). For both assays, the cut-off and the equivocal range were supported by the findings of the clinical study and the correlation to the predicate device assay. N. Proposed Labeling: The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10. {24} O. Conclusion: The submitted information in this premarket notification is complete and supports a substantial equivalence decision. 25