Kam Kai Man, Luey Cindy Kit Yee, Tsang Yee Man, Law Choi Ping, Chu Man Yu, Cheung Tze Leung, Chiu Agatha Wai Huen
Public Health Laboratories, Public Health Laboratory Centre, Hong Kong Department of Health, Shek Kip Mei, Kowloon, Hong Kong.
J Clin Microbiol. 2003 Oct;41(10):4502-11. doi: 10.1128/JCM.41.10.4502-4511.2003.
Two hundred twenty isolates of Vibrio cholerae O1 and O139 collected from 1994 to 2002 in Hong Kong were analyzed by pulsed-field gel electrophoresis (PFGE). Chromosomal DNAs from all V. cholerae isolates in agarose plugs were digested with the restriction enzyme NotI, resulting in 20 to 27 bands. Sixty distinctive PFGE patterns in the range of 10 to 300 kb were noted among 213 isolates typeable by PFGE. By comparing the common PFGE patterns obtained from four well-defined outbreaks of V. cholerae O1 and O139 with those obtained from other, epidemiologically unrelated isolates during the study period, indistinguishable and similar PFGE patterns were identified, indicating their close relatedness, in agreement with the results of epidemiological investigations. Heterogeneous PFGE patterns (with four to six banding differences), however, were identified among strains that were imported from other parts of Asia, including Indonesia, India, and Pakistan. Correlations with epidemiological information further support the usefulness of PFGE as an epidemiological tool in laboratory investigations of suspected outbreaks. Standardization of PFGE methodology will allow international comparison of fingerprint patterns and will form the basis of a laboratory network for tracking V. cholerae.
采用脉冲场凝胶电泳(PFGE)技术,对1994年至2002年期间从香港收集的220株霍乱弧菌O1和O139菌株进行了分析。用限制性内切酶NotI消化琼脂糖块中所有霍乱弧菌菌株的染色体DNA,产生20至27条带。在213株可用PFGE分型的菌株中,发现了60种独特的PFGE图谱,大小在10至300kb之间。通过比较从4起明确的霍乱弧菌O1和O139疫情中获得的常见PFGE图谱与研究期间从其他流行病学无关的菌株中获得的图谱,鉴定出了无法区分和相似的PFGE图谱,表明它们密切相关,这与流行病学调查结果一致。然而,在从亚洲其他地区(包括印度尼西亚、印度和巴基斯坦)输入的菌株中,发现了异质性PFGE图谱(有四到六个条带差异)。与流行病学信息的相关性进一步支持了PFGE作为疑似疫情实验室调查中流行病学工具的有用性。PFGE方法的标准化将允许对指纹图谱进行国际比较,并将成为追踪霍乱弧菌的实验室网络的基础。