摘要
用T4噬菌体表达的重组新城疫病毒HN蛋白(rHN)为包被抗原,建立新城疫抗体检测的酶联免疫吸附实验(rHN-ELISA).结果表明,抗原最适包被浓度为1.6μg/100μL,待检血清最适稀释度为1:80,鸡血清样品的光吸收值OD450大于0.126可判定为阳性结果,而SPF鸡血清,非免疫鸡血清以及MD,IBD,IB,ILT,AI阳性血清的检测结果均为阴性.对批内和批间样品重复检测的变异系数分别为3.7%～8.5%和3.1%～7.4%.人工感染NDV的SPF鸡第3 d即可用rHN-ELISA检测到抗体,灵敏度明显高于HI和AGP试验.因此,rHN-ELISA是1种特异、敏感、快速的ND抗体检测方法.
The recombinant HN protein （rHN） of Newcastle disease virus （NDV）, expressed by T4 phage, were used to develop an enzyme - linked immunosorbent assay （ rHN - ELISA） to detect antibody titers of Newcastle disease （ND）. The results showed that the optimum amount of rHN antigen coated onto the polystyrene microtieration plate was 1.6 μg/100 μL and appropriate dilution of the serum samples was 1 ： 80. The samples with an optical absorbance value OD450 greater than 0. 126 were judged as positive based on the data from 25 serum of SPF chickens. All the serum samples from SPF chickens, non - vaccined chickens and the chickens infected by other pathogens such as MDV, IBDV, IBV, ILTV and AIV, were negative detected by rHN - ELISA. The variant coefficients of OD450 value within assay and among assays were 3.7% -8.5% and 3.1% -7.4%, respectively. Furthermore, the rHN -ELISA was more sensitive than HI and AGP test, and was able to detect the serum ND antibody from chickens infected with the LaSota strain of NDV on the 3^rd day after inoculation. In conclusion, the rHN -ELISA is a specific, sensitive and rapid method for ND antibody detection.
出处
《西南农业大学学报（自然科学版）》
CAS
CSCD
北大核心
2005年第4期509-511,共3页
Journal of Southwest Agricultural University
