以ABTS为底物的双抗体夹心间接ELISA法的建立及应用

Establishment and Application of the Double Antibodies Indirect Sandwich ELISA Choosing ABTS as Substrate

以多-单克隆抗体与正常鼠腹水平行包被固相载体,并用一种新的底物——ABTS作指示系统,建立了EHFV的双抗体间接夹心ELISA法。结果表明:1)ABTS与酶的反应平稳,不需加终止剂终止酶的反应,结果易于肉眼观察;2)对EHFV的定量测定较IFA客观、准确;3)对EHFV抗原的检出范围为351.65±122.82～2091.87±31.27ng/ml;4)对EHFV感染组织和细胞内EHFV的检出率为100％。以上结果说明该法敏感、特异、快速、简便、实用。并指出ABTS作为辣根过氧化物酶的底物的换代产品,应用前景广泛。

A Modified double antibodies indirect sandwich ELISA with poly-monoclonal antibodies and normal mice ascitic parallel coated micto-plate and a new substrate-ABTS,is 2 ,2-azino-di-3-ethyl-benzthia-zoline sulfrate was established. The modified ELISA was successfully applied to detect EHFV antigen. The results as follows :l)The reaction of ABTS with peroxidase is stable, which doesn't need stop agents, and the result easily observed with naked eyesi2)The method is more objective and accurate than the immunofluo-rescence for quantitative analysis of EHFV|3)Detection rate of EHFV antigen is 100% ;4)Detection range of EHFV antigen is 351. 65+122. 82 to 2091. 87+31. 27ng/ml. The above results indicated that the established ELISA is specific, sensitive,rapid,simple and useful. ABTS can be widely used as a new substrate of the horseradish preoxisase.