副溶血性弧菌重复序列-PCR分型研究

Typing of Vibrio parahaemolyticus by Repetitive Element Sequence-based PCR

摘要:

利用基因外重复回文序列-PCR(REP-PCR)和肠细菌基因间共有重复序列-PCR(ERIC-PCR)技术, 对副溶血性弧菌进行了分子分型研究和亲缘关系的探讨, 并使用Hunter和Gaston方法计算分辨力指数。结果显示40株副溶血性弧菌分离株均可扩增产生可重复的DNA指纹图谱, 并且不同菌株基因组DNA的扩增条带具有多态性。根据SPSS10.0软件得出的树状图结果, REP-PCR可以把40株菌分为21个型, 分辨力指数可达到0.953, 优势菌型为G1型; ERIC-PCR可将40株菌分成4个型, 分辨力指数为0.5。研究显示重复序列-PCR方法可以用于该菌分型分析, REP-PCR具有较好的分型能力。在两种PCR的DNA指纹图谱中, 血清型O1群与O3群主条带均非常相似, 表明它们之间亲缘关系密切。

Abstract:

Two kinds of Rep-PCR, Repetitive extragenic palindromic elements-PCR(REP-PCR) and enterobacterial repetitive intergenic consensus sequences-PCR (ERIC-PCR), were used firstly in typing and genetic relationship research for Vibrio parahaemolyticus in China, and Hunter＆Gaston’s method was used to calculate discriminative index of REP-PCR and ERIC-PCR. Forty Vibrio parahaemolyticus strains, representing a wide range of REP and ERIC patterns, were grouped into 21 and 4 patterns with discriminative index of 0.953 and 0.5, by REP-PCR and ERIC-PCR respectively. G1 type was predominant in 40 strains of Vibrio parahaemolyticus when typed by REP-PCR. The results show that Rep-PCR is feasible for typing Vibrio parahaemolyticus strains, and REP-PCR possesses better typing potential. Similar DNA fingerprints were gained from serotype O1 and O3 strains by two kinds of Rep-PCR. This result indicate that the genetype of serovar O3 and O1 are closely related.