动物源弯曲菌分离纯化鉴定方法优化

Optimization of isolation,purification and identification of Campylobacter from animal

目的优化动物源弯曲菌分离纯化鉴定方法。方法采集新鲜样品,选择不同增菌方式获得菌悬液;通过CCDA、弯曲菌显色培养和Skirrow方法分离弯曲菌;生化分析和分子生物学鉴定弯曲菌。与国标方法相比较,评价该法的灵敏度。结果适合于鸡盲肠和动物粪便中弯曲菌的分离纯化鉴定流程为CCDA选择性分离、弯曲菌显色培养基纯化、哥伦比亚血琼脂进行扩增及分子学方法鉴定。使用优化后的方法对东北地区部分鸡屠宰场盲肠样品进行分离,共采集样品200批次,分离鉴定获得空肠弯曲菌共计110株,分离率为55%。结论建立了一套动物源弯曲菌分离纯化鉴定的方法。该方法简单、能有效提高弯曲菌的分离率,解决了弯曲菌分离难,鉴定难,保存难等问题,弥补了国标中的不足,适合于普遍应用及推广。

Objective To optimize the isolation,purification and identification methods of Campylobacter from animals.Methods Fresh samples were collected to obtain bacterial suspension by different enrichment methods;Campylobacter was isolated by CCDA,Campylobacter chromogenic culture and Skirrow method;Campylobacter was identified by biochemical analysis and molecular biology.Compared with the national standard method,the sensitivity of the method was evaluated.Results The suitable procedures for isolation,purification and identification of Campylobacter from chicken cecum and animal feces were CCDA selective separation,Campylobacter chromogenic medium purification,Columbia blood agar amplification and molecular identification.The optimized method was used to separate cecal samples from some chicken slaughterhouses in northeast China,a total of 200 batches of caecum samples were collected,and a total of 110 Campylobacter jejuni strains were isolated and identified,and the isolation rate was 55%.Conclusion A set of method for isolation,purification and identification of Campylobacter from animals was established.The method is simple,and can effectively improve the isolation rate of Campylobacter,solve the problems of difficult separation,identification and preservation of Campylobacter,and make up for the deficiencies in the national standard,which is suitable for general application and promotion.