戊型肝炎病毒抗体双抗原夹心法ELISA的建立与初步应用

目的　建立抗戊型肝炎病毒 (HEV)抗体检测的双抗原夹心ELISA(DS -ELISA) ,并应用于多种动物血清的检测。方法　将大肠杆菌表达的一段HEVORF2区重组抗原分别包被微孔板和进行辣根过氧化物酶 (HRP)标记 ,利用 5份阳性血清和 4 0份阴性血清建立双抗原夹心ELISA ;用 4 0 0份义务献血员血清比较双抗原夹心ELISA与间接法IgG抗体ELISA的符合情况 ;用 3只HEV感染猴系列血清比较双抗原夹心ELISA试剂和Genelabs公司HEVIgG试剂 ;用双抗原夹心ELISA试剂检测新疆地区的部分牛、绵羊、山羊、猪血清和上海地区的部分鸡血清中的HEV抗体。结果　建立了检测HEV抗体的双抗原夹心ELISA方法 ,对 4 0 0份义务献血员血清的检测表明其与间接法IgG抗体ELISA试剂的符合情况良好 ,并且有更高的s/co比值 ;与Genelabs公司HEVIgG试剂的比较表明双抗原夹心ELISA试剂的检出更早 ,尤其是持续时间及强度明显优于Genelabs试剂 ;在所检测的各种动物中均发现了HEV抗体 ,其中猪抗体的阳性率最高 ,表明双抗原夹心ELISA试剂可同时用于不同动物的抗HEV抗体检测。结论　利用大肠杆菌表达的HEV重组抗原建立了双抗原夹心法ELISA ,并可同时用于不同动物的抗HEV抗体检测。

Establishment and primary application of double antigen sandwich ELISA for detection of hepatitis E virus antibody

Aim To establish a double antigen sandwich ELISA(DS ELISA)for detection of anti hepatitis E virus antibody in different animals.Methods A recombinant antigen expressed in E.coli. was used to coate microplate and conjugated with HRP respectively,the conditions of DS ELISA were determined using five HEV antibody positive sera and forty antibody negative sera.The accordance of DS ELISA with indirectly IgG ELISA was compared by paralleling detection of 400 blood donors'sera using both kits.The serial sera of three HEV experimental infected monkeys were detected using the DS ELISA and IgG ELISA kit of Genelabs Co.Some cattle,sheep,goat,swine,and chicken sera were also detected with DS ELISA.Results DS ELISA for detection of anti HEV were established.The results of parallel detection of 400 blood donor's sera by DS ELISA and indirectly IgG ELISA showed that these two kit had good accordance,and the signal/cutoff(s/co)value of DS ELISA was higher than indirectly ELISA.When detecting serial sera of experimental infected monkey,the seraconversion time detected with DS ELISA were much earlier than with a well known commercial IgG ELISA kit,and the antibody's positive period and the titer detected by DS ELISA were better than the commercial kit too.Anti HEV antibodies were found in all kinds of animals detected,the positive rate in swine was the highest.Conclusion A double antigen sandwich ELISA have been established using recombinant antigens expressed in E.coli.,and can be used to detect anti HEV in different kinds of animals.