检测污染兽用疫苗14种支原体PCR方法的建立及应用  

The establishment of the PCR assay for detection of 14 kinds of Mycoplasma contamination in veterinary vaccines

作　　者：英聪[1,2] 王海光[1] 刘灿[1,3] 沈青春[1] 毕丁仁[2] 宁宜宝[1] 

机构地区：[1]中国兽医药品监察所,北京100081 [2]华中农业大学,湖北武汉430070 [3]中国农业大学,北京100193

出　　处：《中国预防兽医学报》2014年第1期42-45,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家948计划项目(2011-G14)

摘　　要：为完善兽用疫苗中支原体污染的检测技术,本研究对污染生物制品的14种常见支原体基因组进行分析,设计一对可以同时检测这14种支原体的通用引物,建立兽用疫苗中支原体污染的PCR检测方法。应用该方法对10种兽用活疫苗的61批次和10种细胞系的21份传代细胞样品进行支原体污染的检测,并与培养法、DNA荧光染色法及成品的支原体PCR检测试剂盒相比较,除成品的支原体PCR检测试剂盒的检测结果不同之外,其他3种方法的检测结果均相同。但培养法耗时长,DNA荧光染色法操作复杂,成品的支原体PCR检测试剂盒只能检测5种污染细胞的常见支原体,出现漏检情况,而本研究建立的PCR检测方法快速、准确,可以为今后兽用疫苗的支原体污染检测工作提供参考。In this study, based on analyzing genomic sequences of 14 kinds of Mycoplasma standard strains which usually contaminated in the bio-products, a PCR detecting assay was established with a pair of universal primers which was able to detect those 14 Mycoplasma contaminated in veterinary vaccines. Using this method, we detected 61 batches of 10 veterinary live vaccines and 21 passage cells of 11 cell lines. Comparison with the bacteria culture assay and DNA fluorescent staining method, the detection results were the same, but less than the commercial PCR kit. However, the commercial PCR kit was only capable to detect 5 kinds of common Mycoplasmas contaminating cells. Otherwise, the culture assay was time-consumed and the DNA fluorescent staining method was somehow complicated. The established PCR detecting assay proved to be specific for the 14 kinds of Mycoplasma which provided useful technique for Mycoplasma detection in veterinary vaccines.

关 键 词：兽用疫苗 支原体污染 PCR检测 

分 类 号：S852.62[农业科学—基础兽医学]