广东地区嗜肺军团菌的扩增片段长度多态性分析

Analysis of Legionella pnumophlia isolated from Guangdong province by amplified fragment length polymorphism protocol

摘要:

【目的】了解广东部分地区2002~2007年环境水中嗜肺军团菌的基因型及遗传学关系。【方法】参考欧洲军团菌感染工作组（the European Working Group for Legionella Infections, EWGLI）制定的分型草案（1.2版），采用PstⅠ酶对我室保存的2株标准菌株及41株不同时间、不同地点的嗜肺军团菌环境分离株进行扩增片段长度多态性（amplified fragment length polymorphism, AFLP）分析，电泳图谱与EWGLI进行比对。【结果】AFLP分型结果稳定，重复性好；43株嗜肺军团菌，其不同来源菌株呈现多态性分布，经AFLP分析得到33个基因型，辨别力指数为99.79%，其中Kingmed AFLP 011为优势基因型。按Dice系数≥0.8，可分为18个群，Kingmed AFLP D群为优势群，占总菌株数的34.88%。由电泳图谱的相似性，初步推断存在EWGLI 001 Lugano型、002 Lugano、012 Rome、013 London、014 London型、015 Dresden型、021 Lyon等7个基因型，以及多个未报道的新基因型。【结论】广东地区嗜肺军团菌的基因型十分丰富，AFLP技术是其分子流行病学研究的有效手段。

Abstract:

[Objective] To analyze the genetic relationship and predominate genotypes among Legionella pneumophila isolated from Guangdong province. [Methods] In total 43 Legionella pneumophila strains were genotyped according to the amplified fragment length polymorphism protocol, and the obtained electropherograms were contrasted to the published standard patterns of the European Working Group for Legionella Infections. [Results] In total 33 genotypes were identified from those 43 Legionella, the discriminative index was 99.79%, and the Kingmed AFLP type no. 011 was predominate. In total 18 clusters were discriminated by 0.8 dice coefficient, and the Kingmed AFLP cluster D was predominate. By comparison, 7 respective genotypes had high similarity to the EWGLI AFLP type no. 001 Lugano, 002 Lugano, 012 Rome, 013 London, 014 London, 015 Dresden and 021 Lyon, and it also existed some potential new genotypes. [Conclusion] The genetic polymorphism of Legionella pneumophila are diverse in the environmental water of Guangdong province, and amplified fragment length polymorphism is one of the effective means for the research of molecular epidemiology.