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Thromb Haemost 2008; 100(05): 929-936
DOI: 10.1160/TH08-05-0305
New Technologies, Diagnostic Tools and Drugs
Schattauer GmbH

Multiplexed genetic profiling of human blood platelets using fluorescent microspheres

Dmitri V. Gnatenko
1   Department of Medicine, State University of New York, Stony Brook, New York, USA
,
Wei Zhu
2   Department of Applied Mathematics and Statistics, State University of New York, Stony Brook, New York, USA
,
Wadie F. Bahou
1   Department of Medicine, State University of New York, Stony Brook, New York, USA
3   Program in Genetics, State University of New York, Stony Brook, New York, USA
› Author AffiliationsFinancial support: This research was supported by grants HL49141 and HL086376 (National Institutes of Health), MP048005 (Department of Defense), NIH Center Grant MOI 10710–5 to the Stony Brook University General Clinical Research Center, and Targeted Research Award (Stony Brook University).
Further Information

Publication History

Received 16 May 2008

Accepted after minor revision 11 August 2008

Publication Date:
22 November 2017 (online)

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Summary

Human platelets have unique and reproducible mRNA profiles, with evidence for distinct profiles in haematopoietic stem cell disorders associated with thrombocytosis. Platelet transcript profiling is traditionally studied by microarray analysis, quantitative reverse transcription-PCR or serial analysis of gene ex-pression,techniques that are labor- and technically-intensive.We have now applied a novel multiplex-based platform for quantitative transcript profiling of human platelets.Simultaneous quantification of 17 platelet transcripts was assayed using intact platelet-rich plasma or gel-filtered platelets lysed in vitro.Accurate and reproducible profiles could be obtained from as few as 5 X 107 platelets (a platelet mass corresponding to ∼100 µl of whole blood), even for the low-abundant platelet transcripts. Correlation coefficients of this 17-member gene set to platelet Affymetrix microarrays were excellent (r2 = 0.949, p < 1 X 10–10), with no correlation to in kind-derived leukocyte profiles, highlighting the cell-specificity of the platform.These data demonstrate that transcript multiplexing using fluorescent micro-spheres can be adapted for rapid molecular profiling using intact platelets (bypassing the need for RNA isolation methods), with potential applicability irrespective of baseline platelet counts.

Keywords

Platelets - transcript profiling - fluorescent microspheres - Luminex - mRNA
 

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