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目的研究探索O139霍乱弧菌杀弧菌抗体的检测方法。方法用微孔板培养和琼脂平板克隆计数相结合的杀弧菌抗体检测方法,对实验菌株及稀释度、补体浓度等关键参数进行筛选;对50份小鼠免疫血清进行杀弧菌抗体滴度检测,并与O139群霍乱弧菌LPS Ig G抗体滴度进行相关分析;对该方法的特异性、线性和精密性进行了验证。结果筛选出最佳菌株为20100603菌株,最佳稀释度倍数为2 000倍,补体最佳稀释倍数为16倍。O139群霍乱弧菌小鼠免疫血清检测到较高的杀弧菌抗体滴度而PBS小鼠免疫血清未检测到杀弧菌抗体滴度。小鼠免疫血清杀弧菌抗体滴度与O139群霍乱弧菌LPS Ig G抗体滴度之间存在正相关关系。验证结果显示,在抑制剂浓度达到1.0~2.0 A600时,抑制率100%;线性回归方程为y=-1.093x+5.058,其相关系数为-0.999,P<0.05;方法批内CV值为15.72%,批间CV值为23.47%。结论初步建立了O139霍乱弧菌杀弧菌抗体的检测方法,该方法具有较高的特异性、线性和精密度。
Objective To investigate the detection of Vibrio cholerae O139 Vibrio cholerae antibody. Methods The detection methods of Vibrio cholerae antibody combined with microplate culture and agar plate clone counting were used to screen the key strains such as experimental strains, dilution and complement concentration. The antibody titer The correlation analysis was carried out with the titer of antibody against V. cholerae LPS Ig G O139. The specificity, linearity and precision of the method were verified. Results The best strain was identified as 20100603, the best dilution was 2000 times, and the optimal dilution of complement was 16 times. O139 group Vibrio cholerae mouse immune sera detected higher antibody titer of Vibrio spp and PBS mouse serum did not detect anti-Vibrio antibody titer. There was a positive correlation between the titer of anti-Vibrio cholerae anti-Vibrio cholerae anti-Vibrio cholerae O139 antibody and the titer of anti-Vibrio cholerae LPS Ig G antibody. The validation results showed that the inhibition rate was 100% when the concentration of inhibitor was 1.0-2.0 A600. The linear regression equation was y = -1.093x + 5.058, the correlation coefficient was -0.999, P <0.05. The intra-assay CV was 15.72 %, CV between batches was 23.47%. Conclusion The detection method of Vibrio cholerae O139 Vibrio cholerae antibody was established preliminarily. The method has high specificity, linearity and precision.