2型鸭疫里氏杆菌抗体间接ELISA检测方法的建立与应用

以2型鸭疫里氏杆菌裂解物作为包被抗原,以辣根过氧化物酶标记的兔抗鸭IgG抗体为第二抗体,以TMB为显色剂,成功地建立了检测鸭血清中2型鸭疫里氏杆菌抗体的间接EL1SA方法。经检测表明该方法特异性强,敏感性比常规的试管凝集试验高320倍。以该方法对免疫2型鸭疫里氏杆菌灭活油乳剂疫苗的试验鸭进行抗体水平的跟踪测定,发现试验鸭于免疫后7d开始产生抗体,28d达到最高峰,然后开始下降,于免疫后56d其抗体水平仍高于免疫后14d的水平。

Detection of the antibody against Riemerella anatipestifer Serotype 2 by indirect enzyme-linked immunosorbent assay

The Riemerella antipestifer (RA)Serotype 2 treated with ultrasonic cracking was used as coated antigen, horseradish peroxidase-labelled rabbit-anti-duck IgG was taken as the second antibody, Tetramethyl Benzidine di hydrochloride (TMB) was used as coloring agent in this test. The indirect enzyme-linked immunosorbent assay (ELISA) was developed for the detection of antibody against RA Serotype 2. This method was proved to be very specific, sensitive and repetitive. The result showed that the ducks treated with inactivated vaccine of RA Serotype 2 had specific antibody in their sera detected with indirect ELISA on the 7th day after vaccination. The titre of the antibody was up to peak on the 28th day after vaccination, and then declines, the titre of the antibody on the 56th day after vaccination was higher than that on the 14th day after vaccination.