新型p1 基因型肺炎支原体分型方法的建立与应用

【目的】针对肺炎支原体新型p1基因型(V2c型)菌株检测工作的需要,建立相应PCR检测方法并进行评价。【方法】针对新型V2c型肺炎支原体菌株p1基因变异区域序列设计特异性扩增引物,建立对V2c型肺炎支原体菌株进行PCR检测的检测方法并用相关基因测序进行验证。使用所建立的巢式多重PCR对北京地区2008-2011年分离到的214株临床肺炎支原体进行分型分析。【结果】特异引物可有效检测出V2c菌株,在其它型别菌株均无阳性扩增。214株肺炎支原体临床分离株中1型菌株占90.2%(193/214),V2a型菌株占0.9%(2/214),V2c型菌株占8.9%(19/214);未检出2型菌株。【结论】针对V2c型肺炎支原体所建立的基于p1基因的PCR检测方法,能有效区分以往方法无法检测出的新型V2c型肺炎支原体菌株,对开展肺炎支原体流行病学调查和病原分析有重要意义。

Development and application of a new p1-based genotyping method for Mycoplasma pneumoniae

Objective]To develop a PCR method for detecting the newly reported genotype(variant 2c,V2c) of Mycoplasma pneumoniae strains.Methods]Specific primer was designed for detecting the V2c type based on the variant region of V2c strain p1 gene.A nested multiple PCR method for V2c strain detection was set up and confirmed by related gene sequencing.In total 214 clinical strains isolated from Beijing between 2008 and 2011 were analyzed by this typing method.Results]Nest multiple PCR typing method is effective to detect the V2c strain.Of the 214 M.pneumoniae strains 90.2%(193/214) were type 1,0.9%(2/214) were variant 2a,and 8.9%(19/214) were V2c.No type 2 was detected.Conclusion]This typing method is effective to distinguish the V2c strains from other variant M.pneumoniae strains,and important for the epidemiological study of Mycoplasma pneumonia infection.