目的了解钢铁企业工业循环冷却塔水中的嗜肺军团菌污染状况及菌株分子生物学特性。方法于2011年3月—2012年9月对邯郸某钢铁厂相对固定的车间冷却塔水进行连续监测,进行嗜肺军团菌的定量分离培养,对分离到的菌株进行血清分型、脉冲场凝胶电泳分型、基因序列分型、毒力基因(lvh、rtx A)检测。结果共检测117份水样,其中20份水样检出嗜肺军团菌,检出率为17.1%;菌落总数为100~50 000 CFU/L,中位数为12 000 CFU/L;共分离到嗜肺军团菌23株,血清型包括Lp1、Lp3、Lp5、Lp6、Lp8,并以Lp1为主(占36.1%,9/23)。23株嗜肺军团菌PFGE分型得到19种不同带型,其中15种带型为国内首次发现。23株嗜肺军团菌分成20个SBT型别,有12个型别为本研究新发现。23株菌中,lvh、rtx A基因阳性的分别有20、22株,两种毒力基因全部阳性的有19株。结论钢铁企业的冷却塔水存在嗜肺军团菌污染,菌型呈现基因多态性,同时具有独特的基因组成,并普遍具有毒力。 Objective To understand the contamination of Legionella pneumophila in the industrial circulating cooling tower of iron and steel enterprises and the molecular biological characteristics of the strains. Methods From March 2011 to September 2012, the relatively fixed plant cooling tower water of a steel plant in Handan was continuously monitored, and the Legionella pneumophila was quantitatively isolated and cultured. The isolated strains were subjected to serotyping and pulsed field coagulation Gel electrophoresis typing, gene sequence typing, virulence genes (lvh, rtx A) detection. Results A total of 117 water samples were detected, of which 20 samples were detected for Legionella pneumophila, the detection rate was 17.1%. The total number of colonies was 100 ~ 50,000 CFU / L with a median of 12 000 CFU / L. To Legionella pneumophila 23 strains, including serotype Lp1, Lp3, Lp5, Lp6, Lp8, and to Lp1 (36.1%, 9/23). Twenty-three strains of Legionella pneumophila PFGE were obtained, of which 15 were found for the first time in China. Twenty-three strains of Legionella pneumophila were divided into 20 SBT types and 12 types were newly discovered in this study. Of the 23 strains, 20 and 22 were positive for lvh and rtx A genes, respectively, and 19 were positive for both virulence genes. CONCLUSIONS Legionella pneumophila contamination exists in the cooling tower water of the iron and steel enterprises. The bacterial strains present genetic polymorphisms, and have unique genetic compositions, and are generally virulent.