预期应用
ELISA法定量测定人血清、血浆或其它相关液体中异常凝血酶原(APT)含量。
介绍
凝血酶原是由肝脏合成的依赖维生素K的凝血因子。异常凝血酶原(Abnormal Prothrombin,APT)和凝血酶原的化学结构极其相似。APT分子氨基端的特定位置上的谷氨酸残基未经羧基化，因而缺乏结合钙离子的结构基础，在一般凝血试验中无凝血活性。原发性肝癌患者血浆APT水平显著升高，APT与肝癌的关系受到关注，其临床应用价值受到重视。
实验原理
本试剂盒应用双抗体夹心酶标免疫分析法测定标本中APT水平。用纯化的抗体包被微孔板，制成固相抗体，往包被单抗的微孔中依次加入APT抗原、生物素化的抗人APT抗体、HRP标记的亲和素，经过彻底洗涤后用底物TMB显色。TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的APT呈正相关。用酶标仪在450nm波长下测定吸光度（OD值），计算样品浓度。
Intended use
This immunoassay kit allows for the specific measurement of human Abnormal prothrombin,APT concentrations in cell culture supernates, serum, and plasma.
Introduction
Abnormal prothrombin was detected by latex agglutination method in the plasma of the patients with acute hepatic failure (AHF) at significantly higher rate (82% of fulminant hepatitis and 100% of subacute hepatitis) than in acute hepatitis (33%). The concentration of abnormal prothrombin was also significantly higher in acute hepatic failure. Since the concentration of abnormal prothrombin reversely correlated with that of hepaplastin test or prothrombin time, the measurement of plasma abnormal prothrombin seemed to be useful in monitoring the severity of acute hepatic injury.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for APT has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any APT present is bound by the immobilized antibody. An enzyme-linked polyclonal antibody specific for APT is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of APT bound in the initial step. The color development is stopped and the intensity of the color is measured.
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