Laroche Maxime, Dunois Claire, Vissac Anne Marie, Amiral Jean
a R&D Department , HYPHEN BioMed , Neuville sur Oise , France.
b Scientific Consultant for HYPHEN BioMed , Neuville sur Oise , France.
Platelets. 2017 May;28(3):235-241. doi: 10.1080/09537104.2016.1265925. Epub 2017 Jan 19.
Functional and genetic assays for measuring platelet microvesicles (PMVs) are presented and discussed. Functional assays concern two groups of methods: a) homogeneous assays using the cofactor activity of phospholipids (PPLs) contained in PMVs and present in assayed plasmas, and a coagulation or a thrombin generation assay (TGA) as "end points"; b) capture-based assays, in which PMVs bind to an immobilized ligand, such as Annexin V in the presence of calcium, or monoclonal antibodies (MoAbs) specific for membrane proteins. Genetic assays aim to detect micro-RNA (miRNA) present in PMVs: miRNA must be extracted from plasma, and the expression pattern can be determined by various methods such as quantitative real-time PCR, microarray or sequencing. All these technical approaches introduce new exploration tools for measuring or quantitating PMVs or their associated activities, as biomarkers for disease evolution, their diagnosis or prognosis, and for monitoring of some antithrombotic or anti-inflammatory therapies. They offer invaluable analytical tools for research, drug discovery and epidemiological studies and have a strong potential as diagnostic tests.
本文介绍并讨论了用于测量血小板微囊泡(PMV)的功能和基因检测方法。功能检测涉及两类方法:a)均相检测,利用PMV中所含且存在于被测血浆中的磷脂(PPL)的辅因子活性,以及凝血或凝血酶生成检测(TGA)作为“终点”;b)基于捕获的检测,其中PMV在钙存在的情况下与固定化配体(如膜联蛋白V)或针对膜蛋白的单克隆抗体(MoAb)结合。基因检测旨在检测PMV中存在的微小RNA(miRNA):miRNA必须从血浆中提取,其表达模式可通过多种方法确定,如定量实时PCR、微阵列或测序。所有这些技术方法都引入了新的探索工具,用于测量或定量PMV或其相关活性,作为疾病进展、诊断或预后的生物标志物,以及用于监测某些抗血栓或抗炎治疗。它们为研究、药物发现和流行病学研究提供了宝贵的分析工具,并且作为诊断测试具有强大的潜力。