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| 牛副流感病毒3型3种基因型多重RT-PCR检测方法的建立 | |
| 引用本文： | 杨帆,石顺利,徐娜,雷宇,常塔娜,李平安,关平原.牛副流感病毒3型3种基因型多重RT-PCR检测方法的建立[J].中国畜牧兽医,2018,45(1):32-38. |
| 作者姓名： | 杨帆 石顺利 徐娜 雷宇 常塔娜 李平安 关平原 |
| 作者单位： | 1. 内蒙古农业大学兽医学院, 呼和浩特 010018; 2. 农业部动物疾病临床诊疗技术重点实验室, 呼和浩特 010018; 3. 内蒙古自治区通辽市家畜繁育指导站, 通辽市 028000 |
| 基金项目： | 内蒙古自治区自然科学基金项目"内蒙古地区牛副流感的分子流行病学研究"（2016MS347） |
| 摘 要： | 为建立检测牛副流感病毒3型（bovine parainfluenza virus type 3,BPIV3）3种基因型的多重RT-PCR方法,根据GenBank上发表的BPIV3 3种基因型病毒株的HN基因序列设计特异性引物,优化反应体系建立多重RT-PCR方法。结果显示,方法可同时扩增出BPIV3 A型150 bp、B型253 bp和C型342 bp的特异性片段,与牛传染性鼻气管炎病毒（IBRV）、牛呼吸道合胞体病毒（BRSV）、牛病毒性腹泻病毒（BVDV）、小反刍兽疫病毒（PPRV）、牛支原体、牛布鲁氏菌、羊布鲁氏菌、牛源多杀性巴氏杆菌A型和B型均无交叉反应,A、B、C基因型BPIV3最低阳性质粒检测量分别为0.89×104、0.92×104和1.53×104拷贝/μL。本试验建立的多重RT-PCR检测方法操作方便、特异性强,应用于临床样本的检测,可快速检测BPIV3 3种基因型。 |
| 关 键 词： | 牛副流感病毒3型(BPIV3) 基因型 多重RT-PCR 检测 |
| 收稿时间： | 2017-07-10 |
Establishment of a Multiplex RT-PCR Detection Method for Three Genotypes of Bovine Parainfluenza Virus Type 3 |
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| YANG Fan,SHI Shunli,XU Na,LEI Yu,CHANG Tana,LI Pingan,GUAN Pingyuan.Establishment of a Multiplex RT-PCR Detection Method for Three Genotypes of Bovine Parainfluenza Virus Type 3[J].China Animal Husbandry & Veterinary Medicine,2018,45(1):32-38. | |
| Authors: | YANG Fan SHI Shunli XU Na LEI Yu CHANG Tana LI Pingan GUAN Pingyuan |
| Institution: | 1. College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018, China; 2. Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease, Ministry of Agriculture, Hohhot 010018, China; 3. Livestock Breeding Station of Tongliao City in Inner Mongolia, Tongliao 028000, China |
| Abstract: | In order to develop a multiplex RT-PCR method for detecting three genotypes of bovine parainfluenza virus type 3 (BPIV3), according to the hemagglutinin-neuraminidase protein genomic sequences of three BPIV3 genotypes obtained from GenBank, three pairs of specific PCR primers were designed. A multiplex RT-PCR detecting three genotypes of BPIV3 was established and optimized. The results showed no cross-reactivity with IBRV, BRSV, BVDV, M.bovis, PPRV, B.melitensis, B.abortus, bovine P.multocida serotype A and bovine P.multocida serotype B. In this study, the amplification produced a series of specific fragments with lengths of 150 bp (BPIV3a), 253 bp (BPIV3b) and 342 bp(BPIV3c), respectively. The limit detection of three recombinant plasmids were 0.89×104, 0.92×104 and 1.53×104 copies/μL. This multiplex RT-PCR method was high specificity and simplicity of operator. It was applied to detect clinical samples and could rapidly detect three genotypes of BPIV3. |
| Keywords: | bovine parainfluenza virus type 3 (BPIV3) genotypes multiplex RT-PCR detection |
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