Molecular epidemiological studies of Vibrio cholerae in Taiwan: genotyping by polymerase chain reaction and DNA sequencing.

D P Liu, P J Chen, C S Lin, T N Wu
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Abstract

To type the Vibrio cholerae strains isolated from sporadic and epidemic cases in Taiwan, 28 toxigenic isolates were studied by sequencing polymerase chain reaction-amplified cholera toxin gene (ctx) fragments. Based on specific base substitutions on positions 115 and 203 of ctxB and comparison with previously published typing system from Centers for Disease Control and Prevention (Olsvik theta et al., J Clin Microbiol 1993; 31:22-5, Ref.1), two genotypes were identified. Cholera strains from imported seafood and sporadic cases in Taiwan had ctxB polymorphism of genotype 1; strains from patients in the 1962 Taiwan epidemic and Taiwan raised soft-shell turtles had ctxB polymorphism of genotype 3. Moreover, one toxigenic non-O1 strain was found to have other 11 different nucleotides in ctxB compared with those of the O1 and O139 strains. Therefore, DNA sequencing is a useful method for obtaining more complete genetic information. The approach could be improved by applying it to other more polymorphic regions of bacterial genome to obtain better epidemiological information among infected cases.

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台湾霍乱弧菌分子流行病学研究:聚合酶链反应基因分型及DNA测序。
为了对台湾省散发性和流行病例分离的28株霍乱弧菌进行分型,采用聚合酶链反应扩增霍乱毒素基因(ctx)片段测序法对产毒株进行了分型研究。基于ctxB位点115和203的特异性碱基替换,并与美国疾病控制与预防中心先前公布的分型系统进行比较(Olsvik theta et al., J clinical microol 1993;31:22-5, Ref.1),鉴定出两个基因型。进口海产霍乱菌和台湾散发性病例呈ctxB基因1型多态性;从1962年台湾流行病例和台湾饲养的软壳龟中分离的菌株具有ctxB基因3型多态性。此外,与O1和O139菌株相比,发现一种产毒非O1菌株在ctxB中具有另外11种不同的核苷酸。因此,DNA测序是获得更完整遗传信息的有效方法。该方法可以通过将其应用于细菌基因组的其他多态区域来改进,以获得感染病例中更好的流行病学信息。
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