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用聚合酶链反应-限制性片段长度多态性方法鉴别猪链球菌1/2型与2型以及1型与14型

Resolution of Streptococcus suis Serotypes 1/2 versus 2 and 1 versus 14 by PCR-Restriction Fragment Length Polymorphism Method.

作者信息

Matiasovic Jan, Zouharova Monika, Nedbalcova Katerina, Kralova Natalie, Matiaskova Katarina, Simek Bronislav, Kucharovicova Ivana, Gottschalk Marcelo

机构信息

Veterinary Research Institute, Brno, Czech Republic

Veterinary Research Institute, Brno, Czech Republic.

出版信息

J Clin Microbiol. 2020 Jun 24;58(7). doi: 10.1128/JCM.00480-20.

Abstract

is an important pathogen of pigs but is also transmissible to humans, with potentially fatal consequences. Among 29 serotypes currently recognized, some are clinically and epidemiologically more important than others. This is particularly true for serotypes 2 and 14, which have a large impact on pig production and also on human health. Conventional PCR-based serotyping cannot distinguish between serotype 1/2 and serotype 2 or between serotype 1 and serotype 14. Although serotype 1/2 and serotype 2 have a very similar locus, they differ in a single-nucleotide substitution at nucleotide position 483 of the gene. Similarly, serotypes 1 and 14 have a very similar locus but also differ in the same nucleotide substitution of the gene. Fortunately, this 483G→C/T substitution can be detected by BstNI restriction endonuclease. A PCR-restriction fragment length polymorphism (RFLP) detection method amplifying a fragment of the gene digested by BstNI restriction endonuclease was developed and tested in reference strains of these serotypes and also in field isolates.

摘要

是猪的一种重要病原体,但也可传播给人类,可能产生致命后果。在目前已识别的29种血清型中,有些在临床和流行病学上比其他血清型更为重要。血清型2和14尤其如此,它们对养猪生产以及人类健康都有很大影响。基于常规PCR的血清分型无法区分血清型1/2和血清型2,也无法区分血清型1和血清型14。虽然血清型1/2和血清型2有非常相似的基因座,但它们在基因的第483位核苷酸处存在单核苷酸替换差异。同样,血清型1和血清型14有非常相似的基因座,但在该基因的相同核苷酸替换上也存在差异。幸运的是,这种483G→C/T替换可以通过BstNI限制性内切酶检测到。开发了一种PCR-限制性片段长度多态性(RFLP)检测方法,该方法扩增由BstNI限制性内切酶消化的基因片段,并在这些血清型的参考菌株以及田间分离株中进行了测试。

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