细胞支原体巢式PCR检测方法的建立及应用  

Establishment and Application of Nested PCR Method for Detecting Mycoplasma Contamination in Cultured Cells

作　　者：黄海军 高其双 彭霞 周莉 向敏 陈志华 占才耀 卢顺 陶弼菲 钱运国 童伟文 李杰 华娟 王莲芳 夏瑜 任远 

机构地区：[1]武汉市畜牧兽医科学研究所,武汉430208

出　　处：《湖北农业科学》2014年第3期690-693,共4页Hubei Agricultural Sciences

基　　金：国家自然科学基金项目(31201938);武汉市晨光计划项目(201150431078);武汉市农科院青年英才计划项目(YC201101);武汉市农科院科技攻关项目(Y1201208)

摘　　要：动物生物制品生产过程中细胞(特别是传代细胞)被支原体污染是个世界性问题。细胞支原体污染可显著影响生产用细胞的培养及生物功能的发挥,最终影响生物制品的生产,给生物制品企业造成巨大损失,建立高效、快速、敏感度高的检测方法十分必要。建立了一种支原体巢式PCR检测方法,该方法高效、快速、灵敏,可以最大限度降低假阴性和假阳性结果的出现。该方法的建立对培养细胞、动物疫苗等生物制品企业的检测标准化管理有一定的借鉴意义。Cell Mycoplasma contamination in the process of producing animal biological products（especially the passage cell line） is a worldwide problem. Cell Mycoplasma contamination can exert a negative influence on culture and display of biological function of cells for production, ultimately affect producing biological products, bring huge losses to the biological products enterprises. Therefore it is necessary to establish an efficient, rapid, sensitive method for detecting cell isolation and culture method. A nested PCR method of detecting Mycoplasma is established, which is efficient, rapid, sensitive and can minimize the appearance of the false negative and false positive results. The establishment of the method have certain significance reference for standardized management of biological products enterprises engaging in cell culture and animal vaccines.

关 键 词：支原体 细胞培养 巢式PCR 生物制品 

分 类 号：S858.28[农业科学—临床兽医学] Q95[农业科学—兽医学]