The development of methods in the
diagnostics of heparin-induced trombocytopenia
Andelová K.1, 2, Minář J.1, 3, Králová S.4, Hrabcová R.5, Radina M.1, Kučerová M.1
1Oddělení klinické biochemie, Onkologické centrum J. G. Mendela, Nový Jičín 2Ústav klinické biochemie, Jesseniova lékařská fakulta, Univerzita Komenského v Bratislavě, Martin 3Ústav potravinářského inženýrství a chemie, Technologická fakulta, Univerzita T. Bati, Zlín 4Centrum pro trombózu a hemostázu při Onkologickém centru J. G. Mendela, Nový Jičín 5Oddělení hematologie a transfuziologie, Onkologické centrum J. G. Mendela, Nový Jičín |
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Summary:
Objective: Heparin-induced thrombocytopenia (HIT II) is an immune-mediated disorder observed in up to 0.5–5% of
heparin-treated patients. Laboratory testing for the diagnosis of HIT II includes a 14C-serotonin release test, PF4/heparin
enzyme-immunoassay, platelet aggregation and flow cytometric assay with various specificities or sensitivities
for the diagnosis of HIT II. The aim of this work was to develop an HPLC method with electrochemical detection (EC) and
prove the possibility of using a non-radioactive serotonin release test for the diagnosis of HIT II.
Material and Methods: Two groups were tested. First group of patients suspect for HIT II, and second control group with
or without heparin therapy. Serotonin release was analyzed by a newly developed HPLC-EC method.
Results: Analytical performance of this method was satisfactory. The method was linear from 5 to 7000 nmol/l with a limit
of detection of 1 nmol/l. The intra- and inter-assay coefficients of variation were 7.7% and 12%, respectively, for a
concentration of serotonin 1460 nmol/l.
Conclusion: In the present work we describe rapid, non-radioactive, specific and sensitive HPLC-ED methods to
determine serotonin release from the donor platelets.
Key words:
heparin induced thrombocytopenia, serotonin release, HPLC, diagnostics.
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