化学发光单孵多层免疫技术检测粪样中轮状病毒

以酶联免疫吸附分析（ＥＬＩＳＡ）夹心测定粪样中轮状病毒（ＲＶ）实验为模型，将传统又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的粪样上清液和根过氧化物酶（ＨＲＰ）标记抗ＲＶ抗原共同孵育，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，

Chemiluminescence Single Incubation Multilayer Immune Technique Detection of Rotavirus in Excrement

Sandwich ELISA for 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 RV sample(Ag) and HRP labeled anti-RV antiboty(Ab) were added together into microwell coated by anti-RV antibody(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 RV in sample when the amount of Ab and Ab were saturated and their affinites to Ag were assumed to be same.A new method termed chemiluminescence single incubation multilayer immune technique(CL-SIMIT) was established on this porpose,in which HRP catalyzing luminol-p-iodopenol-H_2O_2 reaction was as its detection system.Experiments showed that CL-SIMIT could acheive 4-fold sensitivity of CL-ELISA and 16-fold sensitivity of ELISA,and higher specificity than CL-ELISA tested with several uncorrelational antigens.Detection of 32 samples with ELISA,CL-ELISA,CL-SIMIT showed that the positive ratios were 12/32,12/32,17/32 respectively,while the results of CL-SIMIT accorded well with the clinical dignosis results(18/32).