罕见非嗜肺军团菌的鉴定

Identification of rare non- lungphilic Legionella

目的对非嗜肺军团菌进行分离培养和分子分型鉴定。方法依据《公共场所集中空调通风系统卫生规范》(2006),对广州市海珠区公共场所集中空调冷却塔水进行军团菌检测。用常规分离培养法检测军团菌,进一步运用生化和血清学凝集试验、实时荧光PCR技术及16S rRNA基因、mip基因测序对菌株进行种属鉴定。结果在BCYE平板上生长,在L-半胱氨酸缺失的BCYE平板及血平板中均不生长的菌株,通过实时荧光PCR技术及16S rRNA基因、mip基因测序鉴定为菲氏军团菌4株和釜山军团菌1株,这是本区首次从集中空调冷却塔水中检出的非嗜肺军团菌。结论运用常规分离培养方法和分子生物技术相结合检测出菲氏军团菌和釜山军团菌(国内罕见)等多种军团菌种,同时提示海珠区公共场所集中空调冷却塔水存在非嗜肺军团菌污染;16S rRNA基因、mip基因核酸测序鉴定有助于发现军团菌新种。

Objective To determine the phenotype and genotype of non- lungphilic Legionella. Methods Base on＂Hygienic specifications for air conditioning ventilation systems in public places＂（ 2006）. Legionella were cultivated from the cooling water in central air conditioning system in Haizhu,Guangzhou. Legionella was isolated by culture technique,and then biochemistry test,serotype test,PCR,16 S rRNA and mip gen were performed for the identification. Results The colony grew on the flat of BCYE,did not grow on the BCYE- CYS and blood flat. There were 4 strains of L. feeleii and 1 strain of L. busanensis by PCR,16 S rRNA gene and mip gene sequencing. This is the first non- lungphilic Legionella from cooling tower water of central air conditioning detection in Haizhu. Conclusion L. feeleii and L. busanensis and other Ls have been identified by routine culture and molecular diagnosis test. At the same time,that suggested the cooling tower water of central air conditioning was polluted by non- lungphilic Legionella in Haizhu. nucleic acid sequencing and identifying of 16 S rRNA gene,mip gene are helpful to find new Legionella species.