支原体种属特异性PCR引物的设计及对细胞培养物支原体的检测

Species-specific PCR for Identification of Common Contaminant Mycoplasma in Cell Culture

目的快速全面检测感染细胞、疫苗等多种生物制品中的支原体.方法从多种支原体的16SrRNA核酸序列中选择了两段高度保守的核酸序列,作为支原体种属特异性引物,对标本进行检测.结果用这对种属特异性引物对已知阳性的50份标本进行检测,检测结果显示全部为阳性.应用于实践,检测了89份疫苗标本,阳性率为31.4%,而用常规培养法检测阳性率为12.3%.结论用该种属特异性引物检测支原体具有检测速度快、检测灵敏度高、特异性强等优点.

Objective To detect mycoplasma species mainly infected in cell cultures and vaccinum. Methods PCR primers were designed according to the published sequences of 16 s rRNA of Mycoplasma in genbank, Mycoplasma species were detected by species-specific PCR, Results To detect 50 samples by using speciesspecific PCR, the results are all positive, In practice, we detected 89 vaccinum samples by using speciesspecific PCR method and culture method, 31.4 % and 12.3 % respectively showed the positive. Conclusion The species-specific PCR for mycoplasma could identify mycoplasma with high speed, sensitivity and specification in cell culture.