用于鼠疫疫苗纯菌检查的噬菌体裂解法的建立

Establishment of phage bacteriolytic method for detection of purity of plague vaccines

目的 建立用于鼠疫疫苗纯菌检查的噬菌体裂解法,并与现行的直接接种纯菌检查方法进行比较.方法 使用鼠疫噬菌体裂解在细菌分类学上与鼠疫杆菌相近的细菌及常见的污染细菌,确定鼠疫噬菌体对鼠疫杆菌裂解的特异性.分别将小肠结肠炎耶尔森菌和大肠杆菌定量掺入鼠疫疫苗来制备模拟污染鼠疫疫苗,采用噬菌体裂解纯菌检查方法检测模拟污染鼠疫疫苗,并将检测结果与现行的直接接种纯菌检查方法的检测结果进行比较.结果 鼠疫噬菌体能特异性地裂解鼠疫杆菌,对在细菌分类学上与鼠疫杆菌相近的细菌以及常见的污染细菌无裂解作用.对模拟污染鼠疫疫苗中的小肠结肠炎耶尔森菌和大肠杆菌的检出限,噬菌体裂解法均为1CFU/剂疫苗（50μl疫苗）,现行的直接接种法分别为50和5 CFU/剂疫苗（50μl疫苗）.结论 噬菌体裂解纯菌检查方法具有较好的敏感性和特异性,可用于鼠疫疫苗的质量控制.

Objective To establish phage bacteriolytic method for detection of purity of plague vaccines,and compare the phage bacteriolytic method with the available direct inoculation method.Methods Ten strains of non-Yersinia pestis were lysed with plague phage to evaluate specificity of plague phage.The quantified Yersinia enterocolitica and Escherichia coli were respectively added into plague vaccines to prepare simulated contaminated plague vaccines.The phage bacteriolytic method was used to detect purity of plague vaccines.The detection results between phage bacteriolytic method and available direct inoculation method were compared.Results The plague phage could lyse Yersinia pestis specifically.No cross-reactions were observed between the plague phage and other strains of non-Yersinia pestis.The detection limits of phage bacteriolytic method for Yersinia enterocolitica and Escherichia coli in simulated contaminated plague vaccines were both 1 CFU/dose vaccine （50 μl）.The detection limits of the available direct inoculation method for Yersinia enterocolitica and Escherichia coli in simulated contaminated plague vaccines were 50 and 5 CFU/dose vaccine （50 μl）,respectively.Conclusion The phage bacteriolytic method for detection of purity of plague vaccines has better sensitivity and specificity and can be used for quality control of plague vaccines.