儿童肺泡灌洗液肺炎支原体检出率及基因型分析

汪小五; 丁圣刚; 王林定; 唐振华; 朱义朗; 高勇

doi:10.13898/j.cnki.issn.1000-2200.2019.01.029

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

摘要

摘要:
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%。

Abstract:
ObjectiveTo 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.
MethodsThe 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.
ResultsFifty-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.
ConclusionsThe genotype of epidemic strains of MP is type P1-Ⅰ, and the positive rate of MP infection is 38.2% in Hefei.

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