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【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。
【Objective】 The objective of this study was to determine the need for a new strain of p1 genotype (V2c) of Mycoplasma pneumoniae and to establish a corresponding PCR assay. 【Method】 Aiming at the mutation region of the new type V2c Mycoplasma pneumoniae p1 gene, a specific amplification primer was designed and the PCR detection method of Mycoplasma pneumoniae type V2c was established and verified by sequencing. The established nested multiplex PCR was used to genotype 214 clinical isolates of Mycoplasma pneumoniae from 2008 to 2011 in Beijing. 【Result】 The results showed that V2c was detected by specific primers and no positive amplification was observed in other strains. Among the 214 clinical isolates of Mycoplasma pneumoniae, type 1 strains accounted for 90.2% (193/214), V2a type strains accounted for 0.9% (2/214), V2c type strains accounted for 8.9% (19/214), type 2 strains were not detected . 【Conclusion】 The PCR-based p1 gene-based PCR assay established for Mycoplasma pneumoniae V2c can effectively distinguish the new type V2c Mycoplasma pneumoniae strains that can not be detected by previous methods and is of great significance for the epidemiological investigation and pathogen analysis of Mycoplasma pneumoniae.