摘要
根据GenBank中登录的犬瘟热病毒(CDV)N基因序列,设计合成1对特异性引物,以犬瘟热病毒疫苗中提取的RNA为阳性模板建立了快速检测CDV的RT-PCR方法,结果显示,所建立的CDV RT-PCR扩增得到与理论设计值大小一致的456 bp的特异性片段,对犬细小病毒(PPV)、猪伪狂犬病毒(PRV)、牛轮状病毒(BRV)、牛病毒性腹泻病毒(BVDV)、大肠杆菌(E.coli)和新城疫病毒(NDV)的扩增结果均为阴性;最低可检出约1.53×10~2拷贝/μL的核酸;重复性试验结果表明,该方法检测重复性好.此方法对成都大熊猫繁育研究基地的37份眼观正常的大熊猫粪便样品检测,未检测出犬瘟热病毒,与胶体金试纸卡检测结果一致.
According to the published sequence of canine distemper virus（CDV） N gene in GenBank,a pair of specific primers was designed for the detection of CDV. The template extracted from CDV vaccine was used as positive template to establish RT- PCR detection method for CDV. After optimizing the reaction system,as few as 1.53 × 10^2eopies/μL plasmid DNA of CDV spe- cific fragment could be detected accurately and rapidly. This method had good reproducibility and stability. There were no cross reactions to Canine Parvovirus, Swine Pseudorabies Virus, Bovine Rotavirus, Bovine Viral Diarrhea Virus, E. coil and New Cas- tle Disease Virus. This method was applied for the detection of canine distemper virus in fecal samples of giant pandas, and the results showed that 37 normal giant panda fecal samples collected from Chengdu Research Base of Giant Panda Breeding were CDV RT-PCR negative samples,and the results were consistent with the colloidal gold strip test results.
出处
《西南民族大学学报（自然科学版）》
CAS
2017年第3期237-241,共5页
Journal of Southwest Minzu University(Natural Science Edition)
基金
大熊猫繁育研究基金项目(CPF2015-03)
