A high-performance liquid chromatography method for the serotonin release assay is equivalent to the radioactive method

作者：Sono Koree N K; Crist R A; Frank E L; Rodgers G M; Smock K J^*

来源：International Journal of Laboratory Hematology, 2016, 38(1): 72-80.

DOI：10.1111/ijlh.12442

• Heparin-induced thrombocytopenia
• serotonin release assay
• high-performance liquid chromatography
• platelets
• laboratory testing

IntroductionThe serotonin release assay (SRA) is considered the gold standard laboratory test for heparin-induced thrombocytopenia (HIT). The historic SRA method uses platelets loaded with radiolabeled serotonin to evaluate platelet activation by HIT immune complexes. However, a nonradioactive method is desirable. We report the performance characteristics of a high-performance liquid chromatography (HPLC) SRA method.
MethodsWe validated the performance characteristics of an HPLC-SRA method, including correlation with a reference laboratory using the radioactive method. Serotonin released from reagent platelets was quantified by HPLC using fluorescent detection. Results were expressed as % release and classified as positive, negative, or indeterminate based on previously published cutoffs.
ResultsSerum samples from 250 subjects with suspected HIT were tested in the HPLC-SRA and with the radioactive method. Concordant classifications were observed in 230 samples (92%). Sera from 41 healthy individuals tested negative. Between-run imprecision studies showed standard deviation of <6 (% release) for positive, weak positive, and negative serum pools. Stability studies demonstrated stability after two freeze-thaw cycles or up to a week of refrigeration.
ConclusionThe HPLC-SRA has robust performance characteristics, equivalent to the historic radioactive method, but avoids the complexities of working with radioactivity.

• 出版日期2016-2