[1]罗 俊,刘业兵*,陈 坚,等.双重RT-PCR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,(09):687-690.
 LUO Jun,LIU Ye-bing*,CHEN Jian,et al.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,(09):687-690.
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双重RT-PCR方法检测猪流感病毒和猪繁殖与呼吸综合征病毒
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2010年09期
页码:
687-690
栏目:
诊断和检测技术
出版日期:
2010-09-01

文章信息/Info

Title:
Establishment of a duplex RT-PCR for the detection of swine influenza virus and porcine reproductive and respiratory syndrome virus
文章编号:
1008-0589(2010)09-0687-04
作者:
罗 俊12刘业兵1*陈 坚13宁宜宝1
1. 中国兽医药品监察所,北京 100081;
2. 北京信得威特科技有限公司,北京 100302;
3. 广东永顺生物制药有限公司,广东 广州 511356
Author(s):
LUO Jun12 LIU Ye-bing1* CHEN Jian13 NING Yi-bao1
1. China Institute of Veterinary Drug Control, Beijing 100081, China;
2. Beijing SINDER-VET Technology Co., Ltd. Beijing 100302, China;
3. Guangdong Winsun Bio-Pharmaceutical Co., Ltd., Guangzhou 511356, China
关键词:
猪流感病毒猪繁殖与呼吸综合征病毒双重RT-PCR
Keywords:
swine influenza virus porcine reproductive and respiratory syndrome virus duplex RT-PCR
分类号:
S852.65
文献标志码:
A
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103 TCID50/0.1 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。
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 sensitive and could be detect 103 TCID50/0.1 mL PRRSV and 102 EID50/0.1 mL SIV in a mixed sample. Therefore the duplex RT-PCR assay developed here could be an effective method for rapid detection of clinical SIV and PRRSV.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期:2009-12-23
基金项目:2010年公益性行业(农业)科研专项(201003010-3);“十一五”国家科技支撑计划(2007BAD86B04-4)
作者简介:罗 俊(1983-),男,湖北荆州人,硕士研究生,主要从事动物病毒学研究.
*通信作者:E-mail:liuyebing@ivdc.gov.cn
更新日期/Last Update: 2010-08-30