支原体PCR检测方法的建立和应用

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.

Establishment and Application of a PCR Method for the Detection of Mycoplasma

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.