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支原体PCR检测方法的建立和应用
巢伟,徐雷
0
(湖南省中岸生物药业有限公司)
摘要:
依据GenBank中登录的常见细胞污染支原体16S rRNA基因序列,选择高度保守的区域设计引物,建立了一种快速灵敏的支原体PCR检测方法。该方法可以特异性检测出6种支原体,并能敏感的检测到10个拷贝数的目的基因。采用建立的PCR方法和传统的培养法对23个不同样品(细胞、血清、疫苗成品、半成品)进行检测,符合度100%。该方法的建立可大大缩短疫苗生产过程中支原体检验所需时间。
关键词:  支原体  PCR方法  培养法
DOI:
投稿时间:2012-09-13修订日期:2013-01-07
基金项目:
Establishment and Application of a PCR Method for the Detection of Mycoplasma
巢伟,徐 雷
(Hunan Sinoland Bio-Pharmacy Co., Ltd)
Abstract:
According to the 16S rRNA gene sequence published in GenBank coming from common cell culture contaminant Mycoplasma, a pair of primers from highly conserved nucleotide sequences was designed, and a rapid and sensitive PCR method for detection of Mycoplasma was established. Six reference Mycoplasma strains were correctly detected by PCR method. It can sensitive detect 10 copies templates. The result-agreement rate of culture method and PCR method for 23 different samples (including serum, cell, vaccine products and semi-finished products) were 100%. This method can greatly reduce the detection time of Mycoplasma in vaccine production process.
Key words:  Mycoplasma  PCR method  culture method

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