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一种利用聚合酶链反应和限制性酶切片段长度多态性对禽传染性支气管炎病毒进行分型的新方法。

A new typing method for the avian infectious bronchitis virus using polymerase chain reaction and restriction enzyme fragment length polymorphism.

作者信息

Lin Z, Kato A, Kudou Y, Ueda S

机构信息

Nippon Institute for Biological Science, Tokyo, Japan.

出版信息

Arch Virol. 1991;116(1-4):19-31. doi: 10.1007/BF01319228.

Abstract

Two primers with the length of 22 bases each and 400 bases apart on the spike protein gene of avian infectious bronchitis virus (IBV) were prepared. Using these primers, the genome RNA from twelve strains of the various serotypes were reverse-transcribed to cDNA and amplified by polymerase chain reaction (PCR). With all strains, 400 base DNA was amplified, indicating that there were no apparent insertions or deletions in this region. However, the amplified DNA showed different cleavage patterns by the restriction enzymes. These 12 strains were classified into 5 groups. The strain typing based on a comparison of the cleavage patterns was consistent with the previous serological typing. This study thus provides a simple and rapid method for typing of IBV.

摘要

制备了两条长度均为22个碱基且在禽传染性支气管炎病毒(IBV)刺突蛋白基因上相距400个碱基的引物。使用这些引物,将来自12个不同血清型毒株的基因组RNA逆转录为cDNA,并通过聚合酶链反应(PCR)进行扩增。对所有毒株而言,均扩增出了400个碱基的DNA,表明该区域没有明显的插入或缺失。然而,扩增出的DNA经限制性内切酶消化后呈现出不同的切割模式。这12个毒株被分为5组。基于切割模式比较的毒株分型与先前的血清学分型结果一致。因此,本研究为IBV的分型提供了一种简单快速的方法。

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