A colorimetric, metabolic dye reduction assay detects highly activated platelets: application in the diagnosis of heparin-induced thrombocytopenia. Journal Article

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Authors: Prechel, M. M.; Escalante, V; Drenth, A. F.; Walenga, J. M.
Article Title: A colorimetric, metabolic dye reduction assay detects highly activated platelets: application in the diagnosis of heparin-induced thrombocytopenia.
Abstract: Heparin-induced thrombocytopenia (HIT) is caused by antibodies, elicited in response to heparin anticoagulant therapy, which can cause extreme platelet activation and result in a highly procoagulant state. Most diagnostic tests for HIT antibodies measure in[NON-BREAKING SPACE]vitro platelet activation by detecting aggregation or granule release responses. This study demonstrates that the high level of activation by HIT antibodies leads to a rapid breakdown of platelet metabolic activity. Resting or mildly activated platelets metabolize tetrazolium-based indicator dye to a dark colored product, in proportion to cell concentration and dye incubation time. Highly activated, procoagulant platelets resulting from incubation with HIT antibodies fail to metabolize dye and remain light in color. The loss of ability to metabolically reduce indicator dye provides a colorimetric endpoint that can be used in an in[NON-BREAKING SPACE]vitro washed platelet activation assay to detect HIT antibodies. In a prospective evaluation, 145 diagnostic specimens were tested concurrently by both the colorimetric, dye reduction assay and the clinical laboratory standard, radiolabeled-serotonin release assay ((14)C-SRA). Results were in agreement for 96-100% of cases, depending on the chosen stringency of assay cut-off values. This study demonstrates that the metabolic dye reduction assay is comparable to the (14)C-SRA for HIT diagnosis. In addition, this novel assay may have even wider applicability, facilitating studies on the physiologic and clinical relevance of highly activated platelet populations.
Journal Title: Platelets
Volume: 23
Issue: 1
ISSN: 1369-1635
Publisher: Unknown  
Journal Place: England
Date Published: 2012
Start Page: 69
End Page: 80
Language: English
DOI/URL:

http://ovidsp.ovid.com/ovidweb.cgi?T=JS=Y=N=fulltext=medl=21736539
Notes: ID: 12210; Record Owner: From MEDLINE, a database of the U.S. National Library of Medicine.; Status: MEDLINE; Publishing Model: Journal available in: Print-Electronic Citation processed from: Internet; NLM Journal Code: dsj, 9208117; CAS Registry/EC Number/Name of Substance: 0 (Anticoagulants). 0 (Coloring Agents). 0 (Formazans). 9005-49-6 (Heparin).; Electronic Date of Publication: 20110708; Entry Date: 20120416