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使用脉冲场凝胶电泳(PFGE)和重复序列聚合酶链反应(REP-PCR)分析来自海鲜和人类的单核细胞增生李斯特菌分离株之间的遗传相关性。

Genetic relatedness between Listeria monocytogenes isolates from seafood and humans using PFGE and REP-PCR.

作者信息

Chou Chung-Hsi, Wang Chinling

机构信息

Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi, USA.

出版信息

Int J Food Microbiol. 2006 Jul 15;110(2):135-48. doi: 10.1016/j.ijfoodmicro.2006.02.003. Epub 2006 May 9.

Abstract

Listeria monocytogenes has been isolated from catfish and various non-catfish seafoods. Despite progress that was made to understand the relationship between L. monocytogenes isolated from seafood and humans, no study has emphasized the genetic relatedness between catfish and non-catfish seafood and human isolates. The objectives of this study were to (1) investigate the genetic relationship between L. monocytogenes isolates from catfish, non-catfish seafood and humans using pulsed-field gel electrophoresis (PFGE) and repetitive extragenic palindromic element-based PCR (REP-PCR) linking with serotypes; and (2) compare the distinct capabilities of these methods. The genetic relationship among 36 catfish, 35 non-catfish seafood, and 57 human isolates were analyzed using Bionumerics software. Among a total of 128 isolates, only two subtypes by PFGE with ApaI-digestion, one subtype by PFGE with AscI-digestion and three subtypes by REP-PCR were shared by non-catfish seafood or catfish with human isolates. Although most of the non-catfish seafood and catfish isolates were genetically distinct from human isolates, the human isolates used in this study may not be truly representative of all clinical isolates. Serotype 4b was dominant in human isolates, whereas serotypes 3b, 1/2a, and 1/2b and serotypes 4b, 1/2a and 1/2b were commonly found in catfish and non-catfish seafood, respectively. Furthermore, 97, 87 and 94 subtypes of L. monocytogenes were revealed using the ApaI-digestion, the AscI-digestion, and the REP-PCR, respectively. Their respective discriminatory indexes were 0.994, 0.990 and 0.994. Distinct genetic groups based on the difference of flagella antigens were observed in all three methods. The study suggests that the REP-PCR possesses a similar discriminatory ability as the PFGE for subtyping L. monocytogenes. Therefore, the REP-PCR that is rapid and less expensive could be considered as an alternative method for subtyping L. monocytogenes.

摘要

单核细胞增生李斯特菌已从鲶鱼和各种非鲶鱼海鲜中分离出来。尽管在了解从海鲜中分离出的单核细胞增生李斯特菌与人类之间的关系方面取得了进展,但尚无研究强调鲶鱼和非鲶鱼海鲜与人类分离株之间的遗传相关性。本研究的目的是:(1)使用脉冲场凝胶电泳(PFGE)和基于重复外显子回文元件的PCR(REP-PCR)并结合血清型,研究鲶鱼、非鲶鱼海鲜和人类分离的单核细胞增生李斯特菌之间的遗传关系;(2)比较这些方法的不同能力。使用Bionumerics软件分析了36株鲶鱼、35株非鲶鱼海鲜和57株人类分离株之间的遗传关系。在总共128株分离株中,非鲶鱼海鲜或鲶鱼与人类分离株仅共享两种PFGE ApaI酶切亚型、一种PFGE AscI酶切亚型和三种REP-PCR亚型。尽管大多数非鲶鱼海鲜和鲶鱼分离株在基因上与人类分离株不同,但本研究中使用的人类分离株可能并非所有临床分离株的真正代表。血清型4b在人类分离株中占主导地位,而血清型3b、1/2a和1/2b以及血清型4b、1/2a和1/2b分别常见于鲶鱼和非鲶鱼海鲜中。此外,分别使用ApaI酶切、AscI酶切和REP-PCR揭示了97、87和94种单核细胞增生李斯特菌亚型。它们各自的鉴别指数分别为0.994、0.990和0.994。在所有三种方法中均观察到基于鞭毛抗原差异的不同遗传组。该研究表明,REP-PCR在单核细胞增生李斯特菌亚型分析方面具有与PFGE相似的鉴别能力。因此,快速且成本较低的REP-PCR可被视为单核细胞增生李斯特菌亚型分析的替代方法。

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