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Int J Lab Hematol. 2016 Feb;38(1):72-80. doi: 10.1111/ijlh.12442. Epub 2015 Nov 6.

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

Author information

1
ARUP Laboratories Institute for Clinical and Experimental Pathology, Salt Lake City, UT, USA.
2
Department of Pathology, University of Utah Health Sciences Center, Salt Lake City, UT, USA.
3
Department of Internal Medicine, University of Utah Health Sciences Center, Salt Lake City, UT, USA.

Abstract

INTRODUCTION:

The 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.

METHODS:

We 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.

RESULTS:

Serum 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.

CONCLUSION:

The HPLC-SRA has robust performance characteristics, equivalent to the historic radioactive method, but avoids the complexities of working with radioactivity.

KEYWORDS:

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

PMID:
26542198
DOI:
10.1111/ijlh.12442
[Indexed for MEDLINE]

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