發(fā)布時間：2018-06-13 13:58

  本文選題：BVDV + 二重RT-PCR　； 參考：《中國獸醫(yī)雜志》2017年07期

【摘要】：為了同步檢測牛病毒性腹瀉病毒(BVDV)基因1型和基因2型并能夠加以區(qū)分,根據(jù)Gen Bank中已發(fā)表的BVDV基因1型和基因2型、牛藍舌病病毒(BTV)、口蹄疫病毒(FMDV)、豬瘟病毒(CSFV)、牛傳染性鼻氣管炎病毒(IBRV)的全基因序列,采用Primer Premier 5.0軟件,針對BVDV的高度保守的5'端非編碼區(qū)的核苷酸序列,設(shè)計了兩對特異性引物,二重RT-PCR方法擴增目的片段的大小分別為222 bp和119 bp,經(jīng)過對反應(yīng)體系和條件的優(yōu)化,本研究建立了在同一反應(yīng)體系中鑒別檢測BVDV基因1型和2型的RT-PCR方法,擴增出了與預(yù)期大小相符的目的片段。該方法便捷,特異性高,敏感性強,重復(fù)性好,可作為BVDV分型鑒定和快速診斷的有效方法。
[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.
【作者單位】： 河北農(nóng)業(yè)大學(xué)動物醫(yī)學(xué)院;河北出入境檢驗檢疫局檢驗檢疫技術(shù)中心;河北省獸醫(yī)生物技術(shù)工程技術(shù)研究中心;
【基金】：國家質(zhì)量監(jiān)督檢驗檢疫總局科技計劃項目(2015IK093)
【分類號】：S852.653

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本文編號：2014251