海南地区类鼻疽伯克霍尔德菌PFGE分子分型及同源性分析

Pulsed-field gel electrophoresis molecular typing and homology analysis of Burkholderia pseudomallei in Hainan Province

目的通过对海南地区类鼻疽伯克霍尔德菌分离株进行脉冲场电泳（PFGE）分子分型分析，探讨海南地区类鼻疽伯克霍尔德菌之间的同源性，了解海南地区类鼻疽流行和分布特征。方法对海南地区2006年7月～2013年4月临床分离的97株类鼻疽伯克霍尔德菌采用PFGE方法进行分子分型，用BioNumerics软件进行聚类分析和同源性比较。结果97株类鼻疽伯克霍尔德菌分布于海南省18个市县中的14个市县，其中东方市所占比例最高，为17．5％；沿海地区占90．7％（88／97）。感染类型以败血症为主，占61．9。分离菌的PFGE带型呈现多态性，相似性系数为67．8％。在85％相似度水平以上时，实验菌株分为20个PFGE基因型，其中PFGE12型所占比例最高，占29．8％（29／97），且在本型中有7株菌PFGE条带100％相似，占本型菌的24．1％（7／29）。分离菌中所占比例位于第二和第三的分别是PFGE-9（占18．5％）和PFGE16型（占14．4％）。结论PFGE分子分型可较好地对菌株进行同源性分析。海南地区流行的类鼻疽伯克霍尔德菌PFGE型别呈多态性，但存在一些克隆群优势流行。

Objective To analyze the homology of Burkholderia pseudomallei strains in Hainan Province and to ascer tain its epidemiological characteristics and distribution using pulsed-field gel electrophoresis （PFGE） molecular typing. Methods A total of 97 B. pseudomallei isolates was collected in the Hainan area from July 2006 to April 2013. DNA of the isolates was digested with SpeI and then all 97 isolates were typed using PFGE. Bands were marked with their molecular weight and were then analyzed using BioNumerics software. Results The 97 isolates came from 14 of the 18 cities （counties） in Hainan Province. The City of Dongfang accounted for the highest proportion of isolates （17.5 %）. Isolates were mainly from coastal areas （90.7%）. The type of infection was mainly sepsis, which accounted for 61.9% of infections. The PFGE patterns of B. pseudomallei in Hainan had different genotypes and diversity. The isolates studied had a similarity of 67.8%. Based on a similarity of 85%, the 97 isolates were divided into 20 genotypes. The major genotypes in Hainan were PFGE-12 （29.8%）, PFGE-9 （18.5%）, and PFGE-16 （14.4%）. PFGE-12 had 7 isolates with the same PFGE pattern and these isolates accounted for 24.1 % of the isolates studied. Conclusion PFGE molecular typing can be used to analyze the homology of B. pseudomallei. B. pseudornallei has varied genotypes in Hainan, but some clones are predominant.