OLYMPUS IGG REAGENT

{0} 1 # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY A. 510(k) Number: k073490 B. Purpose for Submission: Addition of plasma (Li-heparin and EDTA) and cerebrospinal fluid (CSF) matrix claim to the predicate device C. Measurand: Immunoglobulin G (IgG) D. Type of Test: Quantitative immunoturbidimetric assay E. Applicant: Olympus America, Inc. F. Proprietary and Established Names: Olympus IgG reagent (OSR6X172) G. Regulatory Information: | Product Code | Classification | Regulation Section | Panel | | --- | --- | --- | --- | | CFN Method, Nephelometric, Immunoglobulins (G, A, M) | Class II | 21 CFR 866.5510 Immunoglobulins A, G, M, D, E Immunological Test System | Immunology (IM82) | H. Intended Use: 1. Intended use(s): System reagent for the quantitative determination of IgG immunoglobulins in human serum and plasma on OLYMPUS analyzers 2. Indication(s) for use: The spectrum of abnormalities in serum immunoglobulin concentration is broad. Abnormal concentrations range from a virtual absence of one or more of the three major classes of immunoglobulins (IgA, IgG and IgM) to polyclonal increases in one or more immunoglobulins. Measurement of these immunoglobulins aids in the diagnosis of abnormal protein metabolism and the body's lack of ability to resist infectious agents. 3. Special conditions for use statement(s): For prescription use only 4. Special instrument requirements: OLYMPUS analyzers: AU400/400ᵉ, 600/640/640ᵉ and 2700/5400 I. Device Description: The device consists of two reagents: R1 buffer (Tris buffer pH 7.2, polyethylene glycol 6000) and R2 (goat anti-IgM antiserum). The reagents contain sodium azide as preservative. {1} 2 J. Substantial Equivalence Information: 1. Predicate device name(s): Olympus IgG reagent (OSR6X45) Roche Tina QUANT IgG Gen. 2 2. Predicate 510(k) number(s): k951013 k050113 3. Comparison with predicate: | Similarities: SERUM/PLASMA | | | | --- | --- | --- | | Item | Device | Predicate | | | Olympus IgG reagent (OSR6X172) | Olympus IgG reagent (OSR6X45) | | Intended Use | System reagent for the quantitative determination of IgG immunoglobulins in human serum, plasma and CSF on Olympus analyzers | Same but in serum only | | Indications for Use | Aid in the diagnosis of abnormal protein metabolism and the body’s lack of ability to resist infection | Same | | Test principle | Immunoturbidimetric | Same | | Antibody | Goat anti-IgG | Same | | Reagent form and storage | Liquid, on-board storage | Same | | On-board reagent stability | 90 days | Same | | Calibrator | Olympus Serum Protein Multi-calibrator | Same | | Calibrator traceability | International Reference Preparation CRM 470 | Same | | Calibration frequency | 90 days | Same | | Expected values | 635-1741 mg/dL | Same | | Differences: SERUM/PLASMA | | | | --- | --- | --- | | Item | Device | Predicate | | | Olympus IgG reagent (OSR6X172) | Olympus IgG reagent (OSR6X46) | | Matrix | Serum, plasma (Li heparin or EDTA) | Serum only | {2} | Similarities: CSF | | | | --- | --- | --- | | Item | Device | Predicate | | | Olympus IgG reagent (OSR6X172) | Roche Tina Quant IgG | | Intended Use | System reagent for the quantitative determination of IgG immunoglobulins in human serum, plasma and CSF on Olympus analyzers | In Vitro test for the quantitative determination of IgG in human serum, plasma and cerebrospinal fluid on Roche/Hitachi Cobas c systems | | Matrix | Serum, plasma (Li-heparin or EDTA), CSF | Same | | Test principle | Immunoturbidimetric | Same | | Antibody | Goat anti-IgG | Same | | Reagent form and storage | Liquid, on-board storage | Same | | Calibrator traceability | International Reference Preparation CRM 470 | Same | | Calibration | Multipoint | Same | | Differences: CSF | | | | --- | --- | --- | | Item | Device | Predicate | | | Olympus IgG reagent (OSR6X172) | Roche Tina Quant IgG | | Expected values | 15-20y, 3.5 mg/dL ± 2.0mg/dL 21-40y, 4.2 mg/dL ± 1.4mg/dL 41-60y, 4.7 mg/dL ± 1.0mg/dL | 1-3 mg/dL | | Instrument required | Olympus AU400/400e, 600/640/640e and 2700/5400 | Roche/Hitachi Cobas c systems | | Reagent on-board stability | Open reagents stable 90 days stored in refrigerated compartment | Open reagents stable 84 days stored in refrigerated compartment | | Calibration frequency | 2 days | As required | # K. Standard/Guidance Document Referenced (if applicable): EN14971 (2000) ISO Medical Devices – Application of Risk Management to Medical Devices; EP7-A2 (2005) CLSI Interference Testing in Clinical Chemistry; EP5-A2 (2004) CLSI Evaluation of Precision Performance of Clinical Chemistry Devices; EP9-A2 (2002) CLSI Method Comparison and Bias Estimation Using Patient Samples; CEN 13640 (2002) Stability Testing of In Vitro Diagnostic Reagents; C28-A2 (2000) CLSI How to Define and Determine Reference Intervals in the Clinical Laboratory; EP6-A (2003) CLSI Evaluation of the Linearity of Quantitative {3} Measurement Procedures: A Statistical Approach; FDA: Draft Guidance document for 510(k) Submission of Immunoglobulins A, G, M, D and E Immunoglobulin Test System In Vitro Devices. ## L. Test Principle: When a sample is mixed with R1 buffer and R2 antiserum solution, human IgG reacts specifically with anti-human IgG antibodies to yield insoluble aggregates. Immune complexes formed in solution scatter light in proportion to their size, shape and concentration. The Olympus analyzer measures the decrease in intensity of light transmitted (increase in absorbance) through particles suspended in solution as a result of complexes formed during the antigen-antibody reaction. ## M. Performance Characteristics (if/when applicable): ### 1. Analytical performance: #### a. Precision/Reproducibility: **Serum/Plasma** Precision is checked, based on CLSI EP5-A2, by testing low, medium and high analyte human serum pools; 2 runs in duplicate for 20 days (n=80) on the AU400/400ᵉ, AU600/640/640ᵉ, and AU2700/5400. The within run precision covering the platforms ranged from 0.88–3.41% and the total precision ranged from 1.51–4.66%. AU400/400ᵉ | N=80 | Within-run | | Total | | | --- | --- | --- | --- | --- | | Mean (mg/dL) | SD | %CV | SD | %CV | | 432 | 4 | 0.88 | 9 | 2.18 | | 1098 | 14 | 1.27 | 25 | 2.29 | | 2237 | 76 | 3.41 | 77 | 3.43 | AU600/640/640ᵉ | N=80 | Within-run | | Total | | | --- | --- | --- | --- | --- | | Mean (mg/dL) | SD | %CV | SD | %CV | | 431 | 5 | 1.14 | 14 | 3.29 | | 1088 | 16 | 1.45 | 38 | 3.49 | | 2173 | 49 | 2.24 | 101 | 4.66 | AU2700/5400 | N=80 | Within-run | | Total | | | --- | --- | --- | --- | --- | | Mean (mg/dL) | SD | %CV | SD | %CV | | 444 | 4 | 0.95 | 7 | 1.51 | | 1060 | 17 | 1.87 | 20 | 1.87 | | 2191 | 26 | 2.03 | 44 | 2.03 | ## CSF Precision is checked, based on CLSI EP5-A2, by testing low, medium and high analyte human serum pools (set to cover the reportable range for CSF); 2 runs in duplicate for 20 days (n=80) on the AU400/400ᵉ, AU600/640/640ᵉ, and AU2700/5400. The within run precision ranged from 0.89–7.93% and the {4} total precision ranged from 2.81-10.24%. AU400/400° | N=80 | Within-run | | Total | | | --- | --- | --- | --- | --- | | Mean (mg/dL) | SD | %CV | SD | %CV | | 3.5 | 0.1 | 2.88 | 0.3 | 9.82 | | 10.2 | 0.1 | 1.44 | 0.4 | 4.08 | | 35.8 | 0.6 | 1.65 | 1.3 | 3.61 | AU600/640/640° | N=80 | Within-run | | Total | | | --- | --- | --- | --- | --- | | Mean (mg/dL) | SD | %CV | SD | %CV | | 3.5 | 0.1 | 2.68 | 0.3 | 9.53 | | 10.1 | 0.1 | 0.89 | 0.4 | 3.67 | | 35.6 | 0.3 | 0.93 | 1.0 | 2.81 | AU2700/5400 | N=80 | Within-run | | Total | | | --- | --- | --- | --- | --- | | Mean (mg/dL) | SD | %CV | SD | %CV | | 3.6 | 0.1 | 7.93 | 0.4 | 10.24 | | 10.1 | 0.1 | 2.91 | 0.6 | 6.29 | | 35.4 | 0.6 | 1.31 | 1.3 | 3.83 | **Auto dilution:** Accuracy: Three auto-dilution samples were diluted manually and run on the instrument. The same samples were diluted automatically by the AU640 and %difference was calculated Accuracy 1:5 | Level | Automatic dilution (mg/dl) | Manual dilution (mg/dl) | % Difference | | --- | --- | --- | --- | | 1 | 3911 | 3680 | -6.3 | | 2 | 3123 | 3051 | -2.4 | | 3 | 1986 | 1906 | -4.2 | Accuracy 1:10 | Level | Automatic dilution (mg/dl) | Manual dilution (mg/dl) | % Difference | | --- | --- | --- | --- | | 1 | 3767 | 3613 | -4.3 | | 2 | 2997 | 2923 | 2.5 | | 3 | 2263 | 2176 | 4.0 | Precision: Three auto-dilution samples were run with the automated dilution protocol on the AU640 to generate 20 replicates per sample. %CVs were calculated. {5} Precision (within run) 1:5 | Level | Mean (mg/dL) | SD (mg/dL) | CV (%) | Essential Specification | | | --- | --- | --- | --- | --- | --- | | 1 | 392 | 4 | 1.01 | ≤4.2% CV | Pass | | 2 | 521 | 6 | 1.11 | | | | 3 | 653 | 7 | 1.08 | | | Precision (within run) 1:10 | Level | Mean (mg/dL) | SD (mg/dL) | CV (%) | Essential Specification | | | --- | --- | --- | --- | --- | --- | | 1 | 197 | 3 | 1.60 | ≤4.2% CV | Pass | | 2 | 256 | 3 | 1.16 | | | | 3 | 322 | 5 | 1.65 | | | b. Linearity/assay reportable range: Serum/Plasma The measuring range for the assay is 75-3000 mg/dL. The procedure used to demonstrate linearity was based on CLSI EP6-A. A series of at least ten analyte concentrations, covering the linear dynamic range was prepared by dilution of a high pool sample. Each dilution was assayed in quadruplicate and the mean analytical results were plotted versus the relative analyte concentrations (% dilution). Studies were performed on the AU400, AU640 and AU2700 analyzers. The acceptance criteria for deviation from the regression line for 75-375 mg/dL and 375-3000 mg/dL ranges were ± 30 mg/dL and ± 8%. The studies showed the assay was linear from 75-3000 mg/dL. There was no high dose hook effect up to 30,000 mg/dL. CSF The measuring range for the assay is 2-50 mg/dL. The procedure used to demonstrate linearity was based on CLSI EP6-A. A series of at least ten analyte concentrations, covering the linear dynamic range was prepared by dilution of a high pool sample. Each dilution was assayed in quadruplicate and the mean analytical results were plotted versus the relative analyte concentrations (% dilution). Studies were performed on the AU400, AU640 and AU2700 analyzers. The acceptance criteria for the deviation from the regression line for 2-5 mg/dL and 5-50 mg/dL ranges were ± 0.5 mg/dL and ± 10% respectively. The studies showed the assay was linear from 2-50 mg/dL. There was no high dose hook effect up to 6,000 mg/dL. c. Traceability, Stability, Expected values (controls, calibrators, or methods): The calibrator is traceable to the International Reference Preparation CRM470 (US designation RPPHS lot 91/0619). Serum/Plasma Reagent on-board stability was demonstrated according to internal procedures where the linearity displayed at day 90 and the % drift from Day 0 from control recovery were calculated. A change of ≤ 8% was demonstrated over the 90 days. Data confirm the calibration stability of 90 days. {6} # CSF Reagent on-board stability was demonstrated according to internal procedures where the linearity displayed at day 90 and the $\%$ drift from Day 0 from control recovery were calculated. A change of $\leq 10\%$ was demonstrated over the 90 days. Data confirm the calibration stability of 2 days for CSF. # d. Detection limit: # Serum/Plasma The Limit of Quantitation (LoQ) for the new assay was determined by testing 3 patient pools, 40 fold at an analyte concentration below the lower end of the measuring range on the AU400, AU640 and AU2700. The analyte level with a CV of less than $20\%$ was determined to be $< 74 \mathrm{mg} / \mathrm{dL}$ . This was determined using a method based on the CLSI protocol EP17-A. | | Mean Concentration (mg/dL) | CV (%) | | --- | --- | --- | | AU400 | 33 | 16.8 | | AU640 | 35 | 6.6 | | AU2700 | 21 | 12.4 | The Limit of Detection (LoD) or the concentration of analyte which is significantly different from zero was determined by testing an analyte free sample twenty-fold on the AU400, AU640 and AU2700. The lowest detectable level was determined to be $\leq 7\mathrm{mg / dL}$ . # CSF The Limit of Quantitation (LoQ) for the new assay (CSF) was determined by testing 3 patient pools, 40 fold at an analyte concentration below the lower end of the measuring range. The analyte level with a CV of less than $20\%$ was determined to be $2\mathrm{mg / dL}$ . This was determined using a method based on the CLSI protocol EP17-A. The Limit of Detection (LoD) or the concentration of analyte which is significantly different from zero was determined by testing an analyte free sample (water) twenty-fold on the AU400, AU640 and AU2700. The lowest detectable level was determined to be $\leq 1\mathrm{mg / dL}$ . # e. Analytical specificity: The impact of bilirubin, lipids and hemoglobin were assessed in accordance with CLSI EP7-A2. Serum/Plasma | Substance | Levels up to | Interference | | | | --- | --- | --- | --- | --- | | | | AU400/400e | AU600/640/640e | AU2700/5400 | | Bilirubin | 40 mg/dL | ≤2% | ≤3% | ≤1% | | Lipids | 1000 mg/dL | ≤3% | ≤5% | ≤2% | | Hemoglobin | 500 mg/dL | ≤3% | ≤3% | ≤3% | | RF | 1200 IU/mL | ≤7% | ≤7% | ≤7% | {7} CSF | Substance | Levels up to | % Interference | | | | --- | --- | --- | --- | --- | | | | AU400/400° | AU600/640/640° | AU2700/5400 | | Bilirubin | 36 mg/dL | ≤ 10% | ≤ 10% | ≤ 10% | | Hemoglobin | 500 mg/dL | ≤ 10% | ≤ 10% | ≤ 10% | f. Assay cut-off: See reference range 2. Comparison studies: a. Method comparison with predicate device: Serum/Plasma | Y method (new) | AU2700 | AU2700/5400 | AU2700/5400 | | --- | --- | --- | --- | | X method (predicate) | AU2700 | AU400 | AU640/640° | | Slope | 0.945 | 1.026 | 0.990 | | Intercept | 37.2 | -23.4 | 5.8 | | Correlation coefficient (r) | 0.998 | 0.996 | 0.998 | | Number of samples | 120 | 120 | 120 | | Range (mg/dL) Y method | 195-2986 | 195-2986 | 195-2986 | | Range (mg/dL) X method | 118-2973 | 177-2906 | 190-2963 | CSF | Y method (new) | AU2700 | AU2700/5400 | AU2700/5400 | | --- | --- | --- | --- | | X method (predicate) | Tina Quant | AU400 | AU640/640° | | Slope | 1.067 | 0.996 | 0.969 | | Intercept | -0.069 | -0.127 | 0.14 | | Correlation coefficient (r) | 0.998 | 0.998 | 0.998 | | Number of samples | 55 | 85 | 86 | | Range (mg/dL) Y method | 2.0-42.9 | 2.1-47.2 | 2.1-47.2 | | Range (mg/dL) X method | 1.98-40.0 | 2.2-47.8 | 2.1-48.6 | b. Matrix comparison: Studies were performed based on CLSI EP9-A2. | Y method | Li-heparin plasma | EDTA plasma | | --- | --- | --- | | X method | Serum | Serum | | Slope | 0.940 | 0.910 | | Intercept | 24.1 | 35.9 | | Correlation coefficient | 0.999 | 0.998 | | Number of samples | 45 | 45 | | Patient mean value – serum mg/dL | 1111.4 | 1111.4 | | Patient mean value – plasma mg/dL | 1068.2 | 1047.2 | | Reference range – serum mg/dL | 214.6-2904.7 | 214.6-2904.7 | | Reference range - plasma mg/dL | 201.2-2581.4 | 202.2-2512.6 | 3. Clinical studies: {8} a. Clinical Sensitivity: Not determined b. Clinical specificity: Not determined c. Other clinical supportive data (when a. and b. are not applicable): Not applicable 4. Clinical cut-off: Not applicable 5. Expected values/Reference range: Expected values may vary with age, sex, diet and geographical location. Serum/Plasma The reference range of 635-1741 mg/dL established for the predicate device was re-verified according to CLSI C28-A2 on the Olympus AU400/AU600 and AU5400. CSF Reference ranges for CSF are 15-20 years: 3.5 mg/dL ± 2.0 mg/dL; 21-40 years: 4.2 mg/dL ± 1.4 mg/dL; and 41-60 years: 4.7 mg/dL ± 1.0 mg/dL. These ranges are based on Tietz Textbook of Clinical Chemistry, 1999. 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.