HEMAGEN CRP 150 KIT (EIA METHOD)

{0} MAR 12 1996 K 940324 # 510(k) Summary Hemagen ® CRP 150 Kit ## 1 Submitter's Name/Contact Person Joseph M. Califano, Manager, Regulatory Affairs ### Address Hemagen Diagnostics, Inc. 34-40 Bear Hill Road Waltham, MA, 02154 Phone: (617) 890-3766 Fax: (617) 890-3748 ### Date Prepared 19 January 1996 ## 2 Device Names Trade Name: Hemagen ® CRP 150 Kit Common Name: C-Reactive Protein Assay (EIA method) Classification Name: C-reactive protein immunological test system (per 21 CFR 866.5270) ## 3 Predicate Device Hemagen ® C-Reactive Protein Kit (EIA method) Reference Docket No: K 944288 Page 1 {1} Page 2 # 510(k) Summary Hemagen ® CRP 150 Kit ## 4 Description of Device An enzyme-linked immunosorbent assay (ELISA) designed for the semi-quantitative detection of C-Reactive Protein (CRP) in human serum and plasma. The ELISA methodology is commonly used for serological evaluations. Antibodies to CRP have been attached to the inner surfaces of the microwell plate. During the initial incubation step, CRP in patient serum binds specifically to the immobilized antibody and remains in place after a wash step. A second antibody, which is conjugated to the enzyme horseradish peroxidase, reacts with CRP bound in the first step. In the wells where the second antibody remains bound, the enzyme catalyzes a color change in the substrate, tetramethylbenzidine (TMB). After the reaction is stopped, the color is read in an EIA plate reader. ## 5 Intended Use of Device This kit has been designed for the measurement of C-reactive protein in human serum and plasma. When used according to instructions, the kit is useful in measuring the acute phase responses in humans to infection, tissue damage and various inflammatory conditions. ### 6.(A) Technological Characteristics The Hemagen CRP 150 Kit is an enzyme-linked immunosorbent assay with the following features: i. The assay's detection range is 1 ug/mL to 50 ug/mL of CRP. ii. A positive control. iii. A set of CRP standards. iv. The HRP conjugate is supplied as a liquid. The predicate device is also an enzyme-linked immunosorbent assay. {2} Page 3 # 510(k) Summary Hemagen ® CRP 150 Kit ## 6.(B) Performance Data ### I. Verification of CRP Standards The CRP Standards have been compared to the W.H.O International Standard for Human C-Reactive Protein (1st International Standard, Code 85/506). A study was conducted to demonstrate the high degree of correlation that exists between the Kit standards and the W.H.O. Standard. ### II. Comparison Testing Eighteen human serum* samples with CRP concentrations ranging from 1 to 100 µg/mL were evaluated concurrently with both the proposed device and the predicate device to demonstrate the equivalence of the CRP concentrations estimated with each of the kits in accordance with the instructions in their respective package inserts. {*In the submission for the predicate device, the EIA methodology was shown to be substantially equivalent to a latex agglutination method by concurrent comparison with 166 human serum samples.} Analysis of the results of this study indicated that a linear relationship with a high degree of correlation exists between the estimated CRP concentrations for assayed serum samples as evidenced by an estimated regression slope of +1.0 and a correlation coefficient of > 0.90. ### III. Assay performance with Serum and Plasma Two sets of plasma samples of 41 samples each from apparently healthy donors were selected. Half of the volume of each sample was converted to serum by recalcification using a standard Ca²⁺/thrombin methodology. All of the plasma and converted serum samples were assayed for CRP concentrations with the Hemagen CRP 150 Kit in accordance with the Draft package insert. The results of the evaluation with proposed device indicate that it can provide accurate estimates of CRP concentrations in both human serum and plasma. {3} 510(k) Summary Hemagen ® CRP 150 Kit ## IV. Interfering Substances Lipemic, hemolytic, and iteric samples were evaluated with the Hemagen CRP 150 Kit following NCCLS Proposed Guideline, "Interference Testing in Clinical Chemistry" Document EP7-P ISSN 0273-3099. Samples with hemoglobin concentrations of ≤ 500 mg/dL, lipid concentrations of ≤ 3,000 mg/dL, and bilirubin concentrations of 20 mg/dL did not significantly affect the assay results. ## V. Precision Studies Between-run reproducibility {Inter Assay} Eight serum samples were assayed five times each, twice a day, on five different days (a total of 50 readings per sample). The results are as follows: | Sample | Mean μg/mL | Std. Deviation | C.V. | | --- | --- | --- | --- | | 1 | 1.4 | 0.2 | 12 | | 2 | 1.7 | 0.2 | 11 | | 3 | 4.0 | 0.3 | 7 | | 4 | 10.0 | 0.8 | 8 | | 5 | 23.6 | 1.9 | 8 | | 6 | 24.9 | 1.9 | 8 | | 7 | 32.5 | 2.2 | 7 | | 8 | 30.2 | 1.6 | 5 | Within-run reproducibility {Intra Assay} Eight serum samples were assayed 19 consecutive times in single runs. The results are as follows: | Sample | Mean μg/mL | Std. Deviation | C.V. | | --- | --- | --- | --- | | 1 | 1.0 | 0.2 | 17 | | 2 | 1.1 | 0.3 | 25 | | 3 | 4.0 | 0.7 | 17 | | 4 | 10.6 | 1.6 | 15 | | 5 | 22.8 | 1.1 | 5 | | 6 | 25.9 | 1.5 | 6 | | 7 | 31.9 | 1.9 | 6 | | 8 | 28.1 | 3.2 | 11 | ## 7 Conclusions The results of the comparative studies support the claim that the Hemagen kit is a safe and effective in vitro diagnostic test and is substantially equivalent to the predicate device.