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目的探讨环境水系军团菌分离培养及其表型和分子分型鉴定方法。方法采用军团菌常规分离培养、生化试验、血清学分型和菌体脂肪酸成分分析、PCR试验、AFLP基因分型和SBT核酸测序分型鉴定等技术方法,对广东地区195份环境水系分离的226株军团菌进行表型和基因型分型鉴定。结果在广东地区195份环境水样中共检出军团菌226株,主要是嗜肺军团菌170株(75.22%),长滩军团菌21株(9.29%)、菲氏军团菌21株(9.29%)、橡树岭军团菌和茴芹军团菌各2株(0.89%),其他非嗜肺军团菌10株(4.42%)。125株军团菌细胞脂肪酸成分测定分嗜肺军团菌、橡树岭军团菌、长滩军团菌、菲氏军团菌等6种类型;54株嗜肺军团菌1型AFLP分型,分为25个基因型。7株不能用生化试验、细胞脂肪酸成分分析和AFLP分型做出鉴定的军团菌,用SBT核酸测序鉴定为菲氏军团菌2株、长滩军团菌3株和茴芹军团菌2株。结论广州地区环境水系军团菌分布以嗜肺军团菌为主,同时有橡树岭军团菌、菲氏军团菌、长滩军团菌和茴芹军团菌等多种军团菌种;SBT核酸测序鉴定有助于发现军团菌新种。
Objective To investigate the isolation and culture of Legionella in environmental water and its phenotypic and molecular typing methods. Methods A total of 226 isolates from 195 environmental waters in Guangdong were isolated and identified by routine isolation and culture, biochemical tests, serological typing, fatty acid composition analysis, PCR test, AFLP genotyping and SBT sequencing. Legionella phenotype and genotyping identification. Results A total of 226 strains of Legionella were detected in 195 samples of environmental water samples in Guangdong, including 170 Legionella pneumophila (75.22%), 21 (9.29%) and 21 (9.29%) Legionella pneumophila, , Legionella oolong and Legionella pneumophila each 2 (0.89%), and other non-Legionella pneumophila 10 (4.42%). 125 strains of Legionella cell fatty acid components were divided into Legionella pneumophila, Oak Ridge Legionella, Long Beach Legionella, Legionella pneumophila and other 6 types; 54 Legionella pneumophila type 1 AFLP typing, divided into 25 genotypes . Seven strains of Legionella that could not be identified by biochemical assay, cell fatty acid composition analysis and AFLP typing were identified as two Legionella pneumophila strains, three Legionella pneumophila strains and two Legionella isolates by SBT DNA sequencing. CONCLUSIONS Legionella pneumophila in the environmental water system of Guangzhou are mainly Legionella pneumophila, and Legionella species such as Oak Ridge Legionella, Legionella pneumophila, Legionella pneumophila and Legionella pneumophila etc. Discover new species of Legionella.