儿童肺泡灌洗液肺炎支原体检出率及基因型分析

The detection rate and genotype analysis of Mycoplasma pneumonie in bronchoalveolar lavage fluid

目的:检测136例儿童肺泡灌洗液标本感染肺炎支原体(MP)的情况并初步了解安徽合肥地区MP基因型情况。方法:采用普通PCR技术从136例儿童的肺泡灌洗液标本筛查出已感染MP的标本,并通过巢式多重PCR技术对MP阳性标本进行基因型分析,对照试验采用FH-MP标准株为DNA模版。巢式多重PCR扩增后的DNA片段构建T载体重组质粒做基因测序验证。结果:普通PCR检测显示有52例MP阳性标本(136例肺泡灌洗液标本)。巢式多重PCR技术检测显示MP阳性标本基因型均为P1-Ⅰ型,而标准株FH-MP的基因型为P1-Ⅱ型。重组质粒经过测序后进行片段对比验证其扩增条带均正确。结论:合肥地区儿童MP流行株基因型为P1-Ⅰ型,感染MP的阳性率为38.2%。

Objective:To detect the distribution of Mycoplasma pneumonie(MP)in bronchoalveolar lavage fluid of 136 children,and preliminarily understand the genotype of MP in Hefei Anhui province.Methods:The MP infection cases in 136 bronchoalveolar lavage fluid samples were screened using PCR.The genotype of MP was analyzed using nest multiplex PCR,and the DNA temple of FH-MP standard strain was set as control.The recombinant plasmid was constructed using the amplified DNA fragment cloned by nest multiplex PCR,and sequenced.Results:Fifty-two positive MP samples were screened in 136 cases of bronchoalveolar lavage fluid using PCR.The nest multiplex PCR results showed that the genotype of MP positive samples was type P1-Ⅰ,and the genotype of standard strain FH-MP was type PCP1-Ⅱ.The sequencing results of the recombinant plasmid showed that the cloned fragment was right.Conclusions:The genotype of epidemic strains of MP is type P1-Ⅰ,and the positive rate of MP infection is 38.2%in Hefei.