Evaluation of Six Erythropoietin Kits

Abstract

SUMMARY. Six erythropoietin (EPa) measurement kits from different manufacturers were evaluated for ease of use and performance. Reference values were compared from patient samples with normal and abnormal circulating serum EPa. The kits evaluated were EPO-Trac RIA, EPORIA, DSL RIA, EPa EIA, IBL ELISA and Medac ELISA (see below for manufacturers' details).The radioimmunoassay (RIA) methods were simple to perform with little preliminary preparation of reagents but the non-isotopic methods were of varying degrees of complexity. Imprecision data showed that performance of the RIA methods was acceptable across the range 4D-150 mUjmL [within-assay coefficient of variation (CY)=2-5%, between-assay CY=3-7%] but poor (CY> 10%) at the lower end of the range (lD-15mUjmL). The IBL enzyme-linked immunosorbent assay (ELISA) kit showed good precision across the range ID-54mUjmL (withinassay CY=5-7%, between-assay CY=8-12%). The performance of the EPa enzyme immunoassay (EIA) and Medac ELISA were acceptable although precision was poor at the lower end of the range ( < 10 m UjmL; within-assay CY 10% and 17% and between-assay CY 13% and 18%, respectively).The ELISA and EIA methods had lower limits of detection (0·4--0'8 mUjmL) than the RIA methods (2·3-3·6mUjmL).Analysis of serum EPa measurement variation between methods showed evidence of significant negative bias with DSL RIA and Medac ELISA when compared with EPO-Trac RIA. Conversely, EPORIA showed significant positive bias and the two remaining methods, EPa EIA and IBL ELISA, showed no evidence of systematic bias when compared with EPO-Trac RIA. Patients with normal circulating concentration of serum EPa gave values that were within the reference range of the kits.

Evaluation of immunoassays for the measurement of erythropoietin (EPO) as an indirect biomarker of recombinant human EPO misuse in sport

“…The precision and accuracy obtained in the present study were similar or better than those obtained in previous studies using the same [8,9,19] or other commercially available kits for EPO measurement [16,18]. Besides, estimated LOD and LOQ for CHEM and ELISA tests were lower than most of the previously reported methods [16][17][18].…”

“…Moreover, EPO measurement has important clinical applications in differential diagnosis of anemia and polycythemia, and monitoring rhEPO administration in EPO deficient patients [18]. Usually, concentration of EPO in serum samples is measured by using immunological techniques [16][17][18][19]. The measurement methods need intralaboratory validation to assure satisfactory intra-and inter-assay precision and accuracy allowing clinically and analytically acceptable results.…”

“…The measurement methods need intralaboratory validation to assure satisfactory intra-and inter-assay precision and accuracy allowing clinically and analytically acceptable results. In addition, in immunological methods, the difference in specificity of the used antibodies may lead to different results when using different assays [16][17][18][19]. Thus, reference ranges for EPO concentration can vary depending on the technique used.…”

“…Different immunoassays exist for measuring EPO [16][17][18][19][20]. In this paper, we report an extended evaluation of the two immunoassays for EPO measurement that were used to develop the mathematical models to detect rhEPO misuse [8,9,11], together with an study on the stability of EPO subjected to freezing/thawing cycles and simulated storage/transportation conditions.…”

Erythropoietin - measurement and clinical applications

“…The results showed that there was a wide variation in results and the only assay that gave comparable results to the calibrator values was the assay that used 87/684 as a reference standard. 30 …”