中国比较医学杂志2017，Vol.27Issue(11)：68-74,7.DOI:10.3969.j.issn.1671-7856.2017.11.014
牛病毒性腹泻病毒RT-PCR方法的建立及应用
Establishment and application of a RT-PCR detection method for bovine viral diarrhea virus
摘要
Abstract
Objective To establish a dual RT-PCR detection method for bovine viral diarrhea virus(BVDV)in bovine-derived samples. Methods The primers were designed and synthesized according to the published BVDV1 and BVDV2 genes containing highly conservative sequences in the 5' untranslated regions(5' UTR)to establish the dual RT-PCR method. The specificity,sensitivity,stability of this method were evaluated. Then 41 bovine-derived samples and 64 bovine plasma samples including bovine calf serum, deproteinized calf serum extract and one lienal polypeptide injection were detected with this method. Results There was no cross reaction with bovine parainfluenza virus type 3(BPIV3), classical swine fever virus(CSFV)and Japanese encephalitis virus(JEV)when samples were detected with the established dual RT-PCR method. The lowest concentration of template DNA for detection of BVDV1 and BVDV2 was 8.87 × 102copies and 6.31 × 102copies per microliter,respectively. Electrophoresis bands of about 151 bp and 303 bp were still amplified and detected after the BVDV1 and BVDV2 cDNA was stored at -30℃ for 12 months. The BVDV positive rate of 41 bovine-derived samples and 64 bovine plasma samples detected with this dual RT-PCR method was 14.6% and 29.7%, respectively. Conclusions The established dual RT-PCR method has the advantages of high efficiency, specificity,sensitivity and stability,and can be used for the detection of BVDV in bovine-derived samples.关键词
牛病毒性腹泻病毒/RT-PCR/牛源样本Key words
Bovine viral diarrhea virus/RT-PCR/Bovine-derived samples分类
医药卫生引用本文复制引用
王吉,贺争鸣,付瑞,李晓波,王淑菁,王莎莎,李威,秦骁,巩薇,岳秉飞..牛病毒性腹泻病毒RT-PCR方法的建立及应用[J].中国比较医学杂志,2017,27(11):68-74,7.基金项目
中国食品药品检定研究院学科带头人培养基金项目(编号:2015X5). （编号:2015X5）
