A simple clot based assay for detection of procoagulant cell-derived microparticles

Rucha Patil; Kanjaksha Ghosh; Shrimati Shetty

doi:10.1515/cclm-2015-0508

Article

A simple clot based assay for detection of procoagulant cell-derived microparticles

Rucha Patil , Kanjaksha Ghosh and Shrimati Shetty

Published/Copyright: October 30, 2015

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From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 54 Issue 5

Abstract

Background:

Cell-derived microparticles (MPs) are important biomarkers in many facets of medicine. However, the MP detection methods used till date are costly and time consuming. The main aim of this study was to standardize an in-house clot based screening method for MP detection which would not only be specific and sensitive, but also inexpensive.

Methods:

Four different methods of MP assessment were performed and the results correlated. Using the flow cytometry technique as the gold standard, 25 samples with normal phosphatidylserine (PS) expressing MP levels and 25 samples with elevated levels were selected, which was cross checked by the commercial STA Procoag PPL clotting time (CT) assay. A simple recalcification time and an in-house clot assay were the remaining two tests. The in-house test measures the CT after the addition of calcium chloride to MP rich plasma, following incubation with Russell viper venom and phospholipid free plasma.

Results:

The CT obtained by the in-house assay significantly correlated with the results obtained by flow cytometry (R²=0.87, p<0.01).

Conclusions:

Though preliminary, the in-house assay seems to be efficient, inexpensive and promising. It could definitely be utilized routinely for procoagulant MP assessment in various clinical settings.

Keywords: cell-derived microparticles; clot based assay; procoagulant activity; Russell viper venom

Corresponding author: Shrimati Shetty, PhD, National Institute of Immunohematology (ICMR), Department of Hemostasis and Thrombosis, KEM Hospital, Parel, Mumbai 400 012, India, Phone: +91 22 24138518, Fax: +91 22 24138521, E-mail: shrimatishetty@yahoo.com

Acknowledgments

The authors would like to thank Council of Scientific and Industrial Research (CSIR) and Department of Science and Technology (DST) Ministry of Science and Technology for funding the study.

Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: None declared.
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

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Received: 2015-6-1

Accepted: 2015-9-15

Published Online: 2015-10-30

Published in Print: 2016-5-1

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Articles in the same Issue

https://doi.org/10.1515/cclm-2015-0508

Keywords for this article

cell-derived microparticles; clot based assay; procoagulant activity; Russell viper venom