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通过聚合酶链反应检测粪便中的牛B组轮状病毒。

Detection of bovine group B rotaviruses in feces by polymerase chain reaction.

作者信息

Chinsangaram J, Akita G Y, Osburn B I

机构信息

Department of Veterinary Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis 95616.

出版信息

J Vet Diagn Invest. 1994 Jul;6(3):302-7. doi: 10.1177/104063879400600304.

Abstract

A pair of primers designed from the sequence of genome segment 9 of group B rat rotavirus (IDIR) were employed to amplify genome segment 9 of a group B bovine rotavirus in a polymerase chain reaction (PCR) and to sequence the derived PCR products. A new pair of primers were synthesized from the obtained sequence data and used in a PCR detection assay for group B bovine rotavirus in fecal samples. In addition, another pair of primers were designed to produce a PCR-derived internal probe. This probe was used in a chemiluminescent hybridization to confirm the specificity and to increase the sensitivity of the assay. This assay could detect 0.1 fg of target double-stranded RNA. It was specific to group B bovine rotavirus and did not detect group B rat (IDIR) and porcine rotaviruses, group A bovine (NCDV), simian (SA-11), equine (H-2), porcine (OSU), human (DS-1), deer, and avian rotaviruses, coronavirus, or other enteric organisms tested in this study.

摘要

从B组大鼠轮状病毒(IDIR)基因组第9节段序列设计的一对引物,用于在聚合酶链反应(PCR)中扩增B组牛轮状病毒的基因组第9节段,并对所得PCR产物进行测序。根据获得的序列数据合成了一对新引物,并将其用于粪便样本中B组牛轮状病毒的PCR检测试验。此外,设计了另一对引物以产生PCR衍生的内部探针。该探针用于化学发光杂交,以确认检测方法的特异性并提高其灵敏度。该检测方法可检测到0.1 fg的目标双链RNA。它对B组牛轮状病毒具有特异性,未检测到B组大鼠(IDIR)和猪轮状病毒、A组牛(NCDV)、猿猴(SA-11)、马(H-2)、猪(OSU)、人(DS-1)、鹿和禽轮状病毒、冠状病毒或本研究中测试的其他肠道生物。

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