马鼻疽补体结合反应检测方法的改进

Improvement of complement fixation reaction method for detection of glanders

为配合国家中长期动物疫病防治规划(2012-2020)对马鼻疽的监测任务,本研究对马鼻疽诊断技术(农业行业标准,NY/T 557-2002)中补体结合试验的体系进行了优化和改进。改进后的方法使用商品化的抗原、补体、溶血素,保证了实验体系的稳定;与原方法相比,该方法将反应的总体系从2.5 m L缩小为0.75 m L,阴阳性血清的稀释度从1∶10降至1∶20,阳性血清、阴性血清、抗原、溶血素、补体、红细胞的使用量均缩减了5倍左右,降低了检测成本,同时将检测用具由试管改为2 m L聚丙烯塑料管,方便了实验室的操作和生物废弃物的无害化处理。利用上述改进的方法和原方法分别对来自河北、北京、天津和上海的47份马血清进行了检测,结果相同,以此验证了两种方法在反应条件和稳定性方面是一致的。

In order to carry out the national long-term animal disease prevention and control planning （2012-2020） monitoring task of the glanders which is a serious animal infectious disease for horse, this study optimized and adjusted the reaction formula of the complement fixation reaction （agricultural industry standard, NY/T 557-2002） for glanders diagnosis. The commercial antigen, complement and hemolysin were used in this improved detection method to ensure the stability of the experimental system. Compared with the original method, in this method, the total reaction system was reduced from 2.5 mL to 0.75 mL, the dilution of the negative and positive serum was decreased from 1：10 to 1：20. The dosages of positive serum, negative serum, antigen, hemolysin, complement, and erythrocyte were decreased by about 5 times. The cost of this method was much less than the old method. At the same time, the testing device was changed from a test tube to a 2 mL polypropylene plastic pipe, which was convenient for the operation of the laboratory and harmless treatment of the biological waste. 47 serum samples of horses from Hebei, Beijing, Tianjin and Shanghai were respectively detected glanders with the improved method and the original method. Both tests showed the same results, which proved that the two methods had the same reaction conditions and stability.