环境水军团菌分离培养及其表型与分子分型研究  

Studies on phenotyping and molecular genotyping of Legionella isolated from environmental water system

作　　者：朱庆义[1] 胡朝晖[1] 赵利伟[1] 陈文聪[1] 杨瑞馥[2] 宋亚军[2] 郭兆彪[2] 

机构地区：[1]广州金域医学检验中心,广东广州510330 [2]中国人民解放军军事医学科学院微生物流行病研究所

出　　处：《中国预防医学杂志》2012年第11期805-810,共6页Chinese Preventive Medicine

基　　金：十一五国家科技重大专项课题资助项目(2008ZX10004-006);国家标准化委员会资助项目(20081021-T-361)

摘　　要：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 determine the phenotype and genotype of Legionella isolated from environmental wa ter system. Methods 195 water samples were collected from 2004 to 2009 for the isolation of Legionella . The bacteriologic examination was performed by culture, biochemical test, seroagglutination test, cellular fatty acid composition analysis, PCR, AFLP and SBT sequencing. Results A total of 226 strains of Legionella were isolated which included 170 strains of L. pneumophila （75.22%）, 21 of L. longbeachae （9.29%）, 211 of I.. feeleii （9.29%） , 2 of L. oakridgensis and 2 of L. anisa （0.89%） and 10 isolates of other non-L. pneumophila （4.42%） . Out of 125 strains of Legionella, 6 types were identified by cellular fatty acid tom position analysis, including L. fineumophila, L. oakridgensis, L. longbeachae, L. feeleli and L. anisa. A total of 25 genotype clusters were determined by AFLP from 54 strains of Legionella serogoup I. There were 7 isolates which could only be typed by SBT sequencing showing 2 strains of L. feeleii, 3 of L. longbeachae and 2 of L. anisa. Conclusions L. pneumophila is the major Legionella in the environmental water system in Guangdong. SBT gene sequencing is able to pick up new types of Legionella .

关 键 词：军团菌 表型 分子分型 环境水 

分 类 号：R378[医药卫生—病原生物学]