Enzymatic Functional Assays of Coagulation Using Small Sample Volumes
Enzymatic Functional Assays of Coagulation Using Small Sample Volumes
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DOI：
10.1093/labmed/lmx067
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发表时间：
2018-02-01
影响因子：
1.3
通讯作者：
Emani, Sitaram
中科院分区：
文献类型：
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作者：
Emani, Sirisha;Nelson, Lisa T.;Emani, Sitaram
目前用于综合血栓形成状态的实验室方法需要大量的血液和较长的周转时间。通过开发新型荧光底物和建立盒上荧光(360/460 nm)检测，在数字微流控平台上实现了抗凝血酶III、蛋白C、凝血因子VIII和纤溶酶原的功能检测。使用取自儿科患者的样本对试剂盒和微量滴定板的结果进行比较。采用线性回归和Bland-Altman图建立相关关系。当在试剂盒上进行试验时，结果与微量滴度板相比没有显著差异。重要的是，所有试剂盒所需的样本体积明显低于微量平板检测方法(L 25亩比2毫升)。这项研究证明了使用小血浆体积进行高保真血栓形成平板检测的可行性。这种接近患者的技术在临床环境中的有效性需要进一步研究。
Current laboratory methods for comprehensive thrombophilia status require large blood volumes and long turn-around times. We demonstrate the feasibility of performing thrombophilia panel testing of enzymatic functional assays on a microfluidic cartridge using low sample volume.Functional assays for Antithrombin III, Protein C, Factor VIII, and plasminogen were adapted on the digital microfluidic platform by developing novel fluorogenic substrates and establishing on-cartridge fluorescence (360/460 nm) detection. Cartridge vs. microtiter plate results were compared using samples obtained from pediatric patients. Linear regression and Bland-Altman plots were used to establish correlations. Results were not significantly different when performed on-cartridge compared to microtiter plates. Importantly, the sample volume required is significantly lower for all on-cartridge compared to microtiter plate assays (25 mu L vs. 2 ml).This study demonstrates the feasibility of thrombophilia panel testing with high-fidelity using small plasma volume. The efficacy of this near-patient technology in clinical settings needs further investigation.