快速ELISA诊断血吸虫病的研究及应用  

STUDY ON RAPID ELISA USING PVC FOR IMMUNODIAGNOSIS OF SCHISTOSOMIASIS AND ITS APPLICATION

作　　者：王秀珍[1] 黎世涛[1] 

机构地区：[1]四川省医学科学院寄生虫病防治研究所

出　　处：《中国血吸虫病防治杂志》1990年第4期31-35,共5页Chinese Journal of Schistosomiasis Control

摘　　要：本研究建立了适用于血吸虫病现场的血清学诊断及血清流行病学调查的一种简便、快速的ELISA方法。其主要特点为:（1）用PVC薄膜代替PS板作载体。（2）抗原预固相于载体上。（3）增大反应系统浓度,缩短孵育时间。（4）用抗人IgGMcAb代替羊抗人IgG制备酶结合物。（5）用无毒底物TMB代替OPD。该法每步仅需孵育5min（37℃）,20min左右即可完成全过程。用该法与经典ELISA对照检测249倒粪孵阳性的血吸虫病人血清,其阳性符合率前者为96.39％（240/249）;后者为97.99％（244/249）。检测150例健康人血清,前者的假阳性率为3.33％（5/150）;后者为2.67％（4/150）,经统计学处理（p>0.05）,两法无显著差异。本研究提供的快速ELISA成套试剂,稳定性和重现性较好,1年内均较稳定。A simple, rapid and easy to handle ELISA is developed in our laboratory to meet the need of sero-immunodiagnosis and seroepidemiology in control programme of schistosomiasis. The main points of modification are as follows : l.Polyvinylchloride (PVC) film with bigger, shallower well is used to replace the usual PS plate as solid support. 2.Egg antigen is fixed on the solid support in advance. 3. The incubation duration is shortened by concentrating the reaction systems. 4. Anti-human IgG McAb is used instead of PcAb with enzyme conjugate. 5. A stable, non-toxic substrate of TMBS is used freshly so as to spare the preparation duration. It takes only 5 minutes for incubation at 37℃ in each step and less then 20 minutes totally in the whole procedure to complete the assay. The positive rate was 96.39% for this rapid ELISA with 249 parasitic identified cases, while being 97.99% with routine method. Among 150 sera samples from normal individuals the false positive rate was 3.33% and 2.67% (p>0.05) for the two methods respectively. A kit of the reagents of this ELISA is provided by this laboratory and is proved to be stable and reproducible when stored for one year.

关 键 词：ELISA诊断 血清学诊断 阳性符合率 健康人血清 工作浓度 结合物 IgG 孵育时间 试剂盒 假阳性反应 

分 类 号：R532.21[医药卫生—内科学]