牛病毒性腹泻病毒基因分型二重RT-PCR检测方法的建立

发布时间：2018-06-13 13:58

本文选题：BVDV + 二重RT-PCR　；参考：《中国兽医杂志》2017年07期

【摘要】：为了同步检测牛病毒性腹泻病毒(BVDV)基因1型和基因2型并能够加以区分,根据Gen Bank中已发表的BVDV基因1型和基因2型、牛蓝舌病病毒(BTV)、口蹄疫病毒(FMDV)、猪瘟病毒(CSFV)、牛传染性鼻气管炎病毒(IBRV)的全基因序列,采用Primer Premier 5.0软件,针对BVDV的高度保守的5'端非编码区的核苷酸序列,设计了两对特异性引物,二重RT-PCR方法扩增目的片段的大小分别为222 bp和119 bp,经过对反应体系和条件的优化,本研究建立了在同一反应体系中鉴别检测BVDV基因1型和2型的RT-PCR方法,扩增出了与预期大小相符的目的片段。该方法便捷,特异性高,敏感性强,重复性好,可作为BVDV分型鉴定和快速诊断的有效方法。
[Abstract]:In order to detect and distinguish the BVDV gene type 1 and gene 2 of bovine viral diarrhea virus (BVDV) gene, the BVDV gene type 1 and gene 2 were reported in GenBank. The whole gene sequence of bovine bluetongue virus (BTV), foot-and-mouth disease virus (FMDV), swine fever virus (CSFV) and bovine infectious rhinotracheitis virus (IBRV) was designed by using Primer Premier 5.0 software. Two pairs of specific primers were designed for the sequence of highly conserved 5'noncoding region of BVDV. The size of the target fragment amplified by double RT-PCR was 222 BP and 119 BP, respectively. After optimizing the reaction system and conditions, a RT-PCR method was established for the identification and detection of BVDV gene type 1 and 2 in the same reaction system. A target fragment corresponding to the expected size was amplified. The method is convenient, specific, sensitive and reproducible. It can be used as an effective method for typing and rapid diagnosis of BVDV.
【作者单位】：河北农业大学动物医学院;河北出入境检验检疫局检验检疫技术中心;河北省兽医生物技术工程技术研究中心;
【基金】：国家质量监督检验检疫总局科技计划项目(2015IK093)
【分类号】：S852.653

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