聚合酶链反应-酶切分型鉴定广州地区环境水源军团菌

Identification of Legionella isolates from environmental water by using PCR combined with enzymatic digestion analysis

【目的】探讨聚合酶链反应-酶切分型在快速鉴定环境水源军团菌方面的应用价值,并了解广州地区环境水源军团菌的分布状况。【方法】对广州地区采集的44份环境水样,作军团菌分离培养,再对分离菌株进行16Sr DNA PCR-酶切分型鉴定、16S rDNA基因测序和mip基因测序鉴定。【结果】在广州地区环境水源分离的112株军团菌,经聚合酶链反应-酶切分型鉴定、16S rDNA基因测序和mip基因测序鉴定,检出嗜肺军团菌66株,非嗜肺军团菌46株,其中菲氏军团菌20株,戈氏军团菌17株,橡树岭军团菌7株,长滩军团菌2株。【结论】聚合酶链反应-酶切分型检测环境水源军团菌是一种简便、快速、特异的鉴定方法;在广州地区环境水源中普遍存在军团菌,主要是嗜肺军团菌,其次是菲氏军团菌,戈氏军团菌,橡树岭军团菌和长滩军团菌。

[ Objective ] To evaluate polymerase chain reaction （PCR） combined with enzymatic digestion for identification of Legionella, and investigate status of Legionella in environmental water systems in Guangzhou. [ Methods] Forty-four water samples collected in Guangzhou were cultivated for LegioneUa, and LegioneUa isolates were identified by PCR- enzymatic digestion, 16S rDNA and mip gene sequencing analysis. [ Resultsl Sixty-six strains of Legionella pneumophila and 46 Non-L. pneumophila were identified by PCR-enzymatic digestion and sequencing analysis. Forty-six strains of Non- L. pneumophila included 20 strains of L. feeilei, 17 L. gormanii, 7 L. oakridgensis and 2 L. longbeachae. [ Conclusion] PCR combined with enzymatic digestion is a simple, rapid, and specific method for the identification of Legionella. L. pneumophila was distributed widely, followed by L. feeilei, L. gormanii, L. oakridgensis and L. longbeachae, in environmental water in Guangzhou area.