血清稀释液的筛选及犬抗狂犬病病毒血清抗体正常值范围的确定

Screening of Serum Dilution and Determination of Normal Range of Antibody against Rabies Virus in Dog Sera

应用间接ELISA法进行抗体检测是评价狂犬疫苗免疫效果的一种快捷有效手段。首先筛选得到一种灵敏、特异的血清稀释液;然后应用其对采集自不同地区的812份未免疫犬血清稀释并进行间接ELISA检测;以OIE标准阴、阳性犬血清为对照,随机选取不同A450范围的犬血清应用RFFIT进行检测;以RFFIT效价低于0.1 IU/ml的犬血清,计算阴性血清正常值范围和95%的可信区间,并建立标准品的回归方程。结果显示：所筛选到的血清稀释液灵敏、特异,有756份血清样品的效价低于0.1 IU/ml,犬抗狂犬病抗体阴性血清的正常值范围为0.127 3±0.059 8,95%的可信区间为0.078 1-0.172 3。应用OIE犬阳性血清标准品建立了A450和抗血清效价的回归方程：A=1.139 IU＋0.470。该试验结果为犬狂犬病防制以及犬狂犬病抗体ELISA检测试剂的研制奠定了基础。

The indirect ELISA for antibody detection is a fast and effective method using in immune effect evaluation of rabies vaccines.In this study,the sensitive,specific and suitable serum dilution was screened.A total of 812 dog serum samples were collected from three regions（Changchun,Nanning and Liuzhou）,and then they were diluted with above screened serum dilution and detected by ELISA method.The positive and negative standard dog sera from OIE were used as controls,and then the sera samples in different A450 ranges were randomly selected and the anti-RV serum titers were detected by RFFIT.The normal range of negative dog sera and 95% confidence intervals were calculated with SPSS10.0 software according to the A450 of dog sera with titers lower than 0.1 IU/ml,and the regression equation was also established based on OIE standards.The results showed that the screened serum dilution was sensitive and specific;at least 756 negative serum samples were obtained;the normal range and 95% confidence interval of negative sera were 0.127 3±0.059 8 and 0.078 1-0.172 3,respectively;and the regression equation was A450=1.139 IU＋0.470.These results lay a foundation for the dog rabies control and the development of ELISA kits for dog antibody against rabies viruses.