高等学校化学学报

高等学校化学学报 ›› 1996, Vol. 17 ›› Issue (2): 219.

• 论文 • 上一篇    下一篇

化学发光单孵多层免疫技术检测粪样中轮状病毒

邹克渭1, 章竹君1, 吕九如1, 赵中媛2   

  1. 1. 陕西师范大学化学系, 西安 710062;
    2. 西安医科大学第一附属医院检验科
  • 收稿日期:1995-03-28 出版日期:1996-02-24 发布日期:1996-02-24
  • 通讯作者: 章竹君
  • 作者简介:邹克渭,男,29岁,讲师.
  • 基金资助:

    国家自然科学基金

Chemiluminescence Single Incubation Multilayer Immune Technique Detection of Rotavirus in Excrement

ZOU Ke-Wei1, ZHANG Zhu-Jun1, LU jiu-Ru1, ZHAO Zhong-Yuan2   

  1. 1. Department of Chemistry, Shaasnxi Normal University, Xi'an 710062;
    2. Medical Lab. Of No.1 Teaching Hospital, Xi'an Medical University
  • Received:1995-03-28 Online:1996-02-24 Published:1996-02-24

PDF (PC)

被引次数

7

摘要： 以酶联免疫吸附分析(ELISA)夹心法测定粪样中轮状病毒(RV)实验为模型,将传统双孵式免疫操作变成单孵式,即在包被RV抗体Ab的微孔中同时加入含RV的粪样上清液和辣根过氧化物酶(HRP)标记抗RV抗体共同孵育,其结果在载体表面形成多层酶免疫复合物,以HRP催化鲁米诺-对碘苯酚-过氧化氢化学发光体系作为最终信号检测系统,从而建立起化学发光单孵多层免疫技术(CL-SIMIT).其灵敏度是ELISA的16倍、CL-ELISA的4倍,分别用ELISA、CL-ELISA和CL-SIMIT对32个样品进行测定,阳性检出率分别为12/32、12/32、17/32.其中CL-SIMIT的测定结果更符合临床诊断结果(18/32).

关键词: 轮状病毒, 酶联免疫吸附分析, 化学发光, 辣根过氧化物酶

Abstract: Sandwich ELISAfor the dtection of Rotavirus(RV)as experimental model,the classical immune procedures were changed from double incubation system(DIS)into single incubation system(SIS) in which RVsample(Ag) and HRPlabeled anti-RV antiboty(Ab*) were added together into microwell coated by anti-RVantibody(Ab).After coincobation,the multilayer complex Ab-Ag-Ab* was formed on the inner surface of microwell.It was illustrated theroically that the enzyme amount in mutilayer complex Ab-Ag-Ab* was linearly related to the concentration of Ag binding site of RVin sample when the amount of Ab and Ab* were saturated and their affinites to Ag were assumed to be same.Anew method termed chemiluminescence single incubation multilayer immune technique(CL-SIMIT) was established on this porpose,in which HRPcatalyzing luminol-p-iodopenol-H2O2 reaction was as its detection system.Experiments showed that CL-SIMITcould acheive 4-fold sensitivity of CL-ELISAand 16-fold sensitivity of ELISA,and higher specificity than CL-ELISAtested with several uncorrelational antigens.Detection of 32 samples with ELISA,CL-ELISA,CL-SIMITshowed that the positive ratios were 12/32,12/32,17/32 respectively,while the results of CL-SIMITaccorded well with the clinical dignosis results(18/32).

Key words: Rotavirus, Enzyme-linked immunosorbent assay, Chemiluminescence, Horseradish peroxidase

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