志贺氏菌RAPD鉴定方法的建立

Establishment of RAPD Identification Method for Shigella

[目的]为志贺氏菌食源性疾病的流行病学溯源和疫情控制提供理论依据。[方法]利用5个随机引物对4种不同来源的志贺氏菌进行RAPD扩增,并对得到的扩增图谱进行聚类分析。[结果]5个随机引物中,引物S3、S5表现出较好的多态性。在引物S5的扩增图谱中,志贺氏菌属中各菌均有1条600 bp左右和1条1 000 bp左右的共同谱带,而利用该引物扩增其他属细菌,扩增谱带很少或没有。这说明引物S5对志贺氏菌属细菌是特异性的。根据引物5的聚类分析结果,可将8株志贺氏菌分成4个类群。除宋内氏志贺氏菌外,其他3种6株菌间的相似系数均为1.00,与传统的血清型分型结果完全一致。[结论]采用引物S5的RAPD反应可用于志贺氏菌的快速检测。

[Objective] The research aimed to provide the theoretical basis for tracking the epidemic origin of Shigella food-borne diseases and its disease control. [ Method] Five random primers were used to make RAPD amplification on 4 different sources of Shigella. And the clustering analysis was made on the obtained amplification patterns. [Result] Among 5 random primers, primers S3 and S5 showed better polymorphism. In the amplification patterns by using primer S5, there were common bands including one band with about 600 bp and one band with about 1 000 bp in each bacteria of Shigella. When this primer was used to amplify other genera bacteria, the amplified bands was few or lack. This indicated that primer S5 was specific for bacteria of Shigella. According to the clustering analysis by using primer S5, 8 strains of Shigella could be divided into 4 groups. Except for S. Sonnei,the similarity coefficients among 6 strains of 3 genera were all 1.00, which was completely accordant with the results of traditional serotype typing. [Conclusion] RAPD reaction by using primer S5 could be used in the fast detection of Shigella.