Abstract：Objective To prepare the standard serum against NDiV and establish a method of detecting NDiV antibody by ELISA so as to monitor the infection status of NDiV in population. Methods NDiV was cultivated by intracerebral inoculation in BALB/c mice aged 3 months, and immunogen was prepared. 6-8 weeks old BALB/c mice were immunized on days 0, 7 and 14 respectively by intraperitoneal injection of the immunogen. The harvested sera were verified for the sensitivity with IFA. C6/36 cells were infected with NDiV to prepare specific NDiV antigen, and ELISA antigen was prepared. The best working concentrations of enzyme conjugate, antigen and the positive serum were titrated. The NDiV ELISA detection technique was established, and the sensitivity, specificity, repeatability and stability were determined. Results The serum titer determined by IFA was 1:320, and there was no cross-reactivity with respiratory syncytial virus and adenovirus. The ELISA detection method was established. The best working concentrations of enzyme conjugate, antigen and positive reference serum were 1:10,000, 2.3 μg/ml and 1:200 respectively. The sensitivity of the ELISA detection was 1:3,200. The inter-assay coefficients of variation of specific antigen and normal antigen were 4.56% and 5.17% respectively, and inter-plate coefficients of variation of specific antigen and normal antigen were 3.38% and 6.64% respectively. The stability test showed that the relative deviation was less than 20%. Conclusions The immunized serum prepared in this study can be used as the standard control serum in NDiV serological detection methods. The established ELISA method has a good sensitivity, specificity, repeatability and stability, and therefore can be used in the serological detection of NDiV.
王德全, 刘渠, 周健明, 梁克峰, 刘晓娜, 朱礼平, 王婧颖. 南定病毒标准血清制备及间接ELISA检测方法的建立[J]. 实用预防医学, 2016, 23(9): 1144-1146.
WANG De-quan, LIU Qu, ZHOU Jian-ming, LIANG Ke-feng, LIU Xiao-na, ZHU Li-ping, WANG Jing-ying. Preparation of standard serum and establishment of indirect ELISA detection method of NDiV antibody. , 2016, 23(9): 1144-1146.
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2016, Vol. 23 
