|
|||||
|
|
| 双重RT-PGR方法检测猪流感病毒和猪繁殖与呼吸综合征病毒 | |
| 引用本文： | 罗俊,刘业兵,陈坚,宁宜宝.双重RT-PGR方法检测猪流感病毒和猪繁殖与呼吸综合征病毒[J].中国预防兽医学报,2010,11(9). |
| 作者姓名： | 罗俊 刘业兵 陈坚 宁宜宝 |
| 作者单位： | 1. 中国兽医药品监察所,北京,100081;北京信得威特科技有限公司,北京,100302 2. 中国兽医药品监察所,北京,100081 3. 中国兽医药品监察所,北京,100081;广东永顺生物制药有限公司,广东,广州,511356 |
| 基金项目： | 2010年公益性行业(农业)科研专项，"十一五"国家科技支撑计划 |
| 摘 要： | 为建立特异、敏感的猪流感病毒(SIV)和猪繁殖与呼吸综合征病毒(PRRSV)的双重RT-PCR检测方法,本研究根据GenBank登录的SIV M基因保守序列和PRRSV美洲型毒株的N基因保守序列,设计合成了2对特异引物,通过对扩增条件的优化,建立检测SIV和PRRSV的双重RT-PCR方法.检测结果显示:该方法可同时扩增出SIV(345 bp)和PRRSv(520 bp)的特异性片段;而猪瘟病毒、猪伪狂犬病病毒、猪细小病毒、猪圆环病毒2型及阴性鸡胚尿囊液核酸扩增结果均为阴性;对SIV和PRRSV 2种病毒混合液的最小检出量分别为102 EID50/0.1 mL和103TCID50/0.1 mL.应用双重RT-PCR和病毒分离法对12份临床疑似样品进行对比检测,结果表明:除双重RT-PCR检测到双阳性的3份混合感染病料中1份未分离到PRRSV外,其余2份均分离出病毒.证明该方法具有良好的特异性、敏感性,可以用于临床样品的早期快速检测. |
| 关 键 词： | 猪流感病毒 猪繁殖与呼吸综合征病毒 双重RT-PCR |
Establishment of a duplex RT-PCR for the detection of swine influenza virus and porcine reproductive and respiratory syndrome virus |
|
| LUO Jun,LIU Ye-bing,CHEN Jian,NING Yi-bao.Establishment of a duplex RT-PCR for the detection of swine influenza virus and porcine reproductive and respiratory syndrome virus[J].Chinese Journal of Preventive Veterinary Medicine,2010,11(9). | |
| Authors: | LUO Jun LIU Ye-bing CHEN Jian NING Yi-bao |
| Institution: | LUO Jun1,2,LIU Ye-bing1,CHEN Jian1,3,NING Yi-bao1 (1. China Institute of Veterinary Drug Control,Beijing 100081,China,2. Beijing SINDER-VET Technology Co.,Ltd.,Beijing 100302,3. Guangdong Winsun Bio-Pharmaceutical Co.,Guangzhou 511356,China) |
| Abstract: | A duplex RT-PCR assay for swine influenza virus (SIV) and porcine reproductive and respiratory syndrome virus (PRRSV) were developed by using two pairs of primers derived from the M gene of SIV and N gene of PRRSV America strain. The assay was shown to specifically amplify a 325 bp fragment from SIV and 520 bp from PRRSV,whereas no PCR products could be amplified from Classical swine fever virus,pseudorabies virus,porcine parvovirus,porcine circovirus 2 and normal chick embryo allantoic fluid. It was sensit... |
| Keywords: | swine influenza virus porcine reproductive and respiratory syndrome virus duplex RT-PCR |
| 本文献已被 CNKI 万方数据 等数据库收录！ | |