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.
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.
References
1. Jy W, Horstman LL, Ahn YS. Microparticle size and its relation to composition, functional activity, and clinical significance. Semin Thromb Hemost 2010;36:876–80.10.1055/s-0030-1267041Search in Google Scholar PubMed
2. Zahra S, Anderson JA, Stirling D, Ludlam CA. Microparticles, malignancy and thrombosis. Br J Haematol 2011;152:688–700.10.1111/j.1365-2141.2010.08452.xSearch in Google Scholar PubMed
3. Laude I, Rongières-Bertrand C, Boyer-Neumann C, Wolf M, Mairovitz V, Hugel B, et al. Circulating procoagulant microparticles in women with unexplained pregnancy loss: a new insight. Thromb Hemost 2001;85:18–21.10.1055/s-0037-1612657Search in Google Scholar
4. Carp H, Dardik R, Lubetsky A, Salomon O, Eskaraev R, Rosenthal E, et al. Prevalence of circulating procoagulant microparticles in women with recurrent miscarriage: a case-controlled study. Hum Reprod 2004;19:191–5.10.1093/humrep/deg512Search in Google Scholar PubMed
5. Patil R, Ghosh K, Satoskar P, Shetty S. Elevated procoagulant endothelial and tissue factor expressing microparticles in women with recurrent pregnancy loss. PLoS One 2013;20:e81407.10.1371/journal.pone.0081407Search in Google Scholar PubMed PubMed Central
6. Patil R, Ghosh K, Damania K, Bansal V, Satoskar P, Darekar A, et al. Effect of anticoagulant therapy on cell-derived microparticles (MP) and pregnancy outcome in women with pregnancy loss. Br J Haematol 2015. [PMID: 25921354] 10.1111/bjh.13460.Search in Google Scholar PubMed
7. Lacroix R, Robert S, Poncelet P, Kasthuri RS, Key NS, Dignat-George F. ISTH SSC Workshop. Standardization of platelet-derived microparticle enumeration by flow cytometry with calibrated beads: results of the International Society on Thrombosis and Haemostasis SSC Collaborative workshop. J Thromb Hemost 2010;8:2571–4.10.1111/j.1538-7836.2010.04047.xSearch in Google Scholar PubMed
8. Shantsila E, Montoro-Garcia S, Gallego P, Lip GY. Circulating microparticles: challenges and perspectives of flow cytometric assessment. Thromb Haemost 2014;111:1009–14.10.1160/TH13-11-0937Search in Google Scholar PubMed
9. Osumi K, Ozeki Y, Saito S, Nagamura Y, Ito H, Kimura Y, et al. Development and assessment of enzyme immunoassay for platelet-derived microparticles. Thromb Haemost 2001;85: 326–30.10.1055/s-0037-1615688Search in Google Scholar
10. Strasser EF, Happ S, Weiss DR, Pfeiffer A, Zimmermann R, Eckstein R. Microparticle detection in platelet products by three different methods. Transfusion 2013;53:156–66.10.1111/j.1537-2995.2012.03720.xSearch in Google Scholar PubMed
11. Chan KS, Sparrow RL. Microparticle profile and procoagulant activity of fresh-frozen plasma is affected by whole blood leukoreduction rather than 24-h room temperature hold. Transfusion 2014;54:1935–44.10.1111/trf.12602Search in Google Scholar PubMed PubMed Central
©2016 Walter de Gruyter GmbH, Berlin/Boston
