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| 猪捷申病毒TaqMan实时定量RT-PCR方法的建立 | |
| 引用本文： | 胡峰,赵亚荣,吕超超,王贵华,崔玉东,王春仁,崔尚金.猪捷申病毒TaqMan实时定量RT-PCR方法的建立[J].中国预防兽医学报,2012,34(3):206-210. |
| 作者姓名： | 胡峰 赵亚荣 吕超超 王贵华 崔玉东 王春仁 崔尚金 |
| 作者单位： | 1. 黑龙江八一农垦大学动物科技学院,黑龙江大庆163319;中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/猪传染病研究室,黑龙江哈尔滨150001 2. 大北农动物医学研究中心,北京,100097 3. 黑龙江八一农垦大学动物科技学院,黑龙江大庆,163319 |
| 基金项目： | 兽医生物技术国家重点实验室基本业务费(SKLVBP201001);大北农横向课题(1167-Z) |
| 摘 要： | 为建立猪捷申病毒(PTV)的早期检测及定量分析方法,本研究基于PTV 11个血清型基因组5′端非编码区保守序列,设计引物和TaqMan探针,建立了检测PTV的TaqMan实时定量RT-PCR方法.应用该方法对PTV、猪细小病毒、猪繁殖与呼吸综合症病毒、猪圆环病毒2型、猪伪狂犬病毒以及猪瘟病毒进行特异性试验,结果除PTV为阳性外其它均为阴性；针对PTV最低可检测到10个拷贝；批内、批间重复试验的变异系数均小于3％.应用建立的方法与病毒分离方法分别对91份临床样品进行检测,检出率分别为79.12％和57.14％,两者的符合率是78.02％.经临床应用表明,该实时定量RT-PCR方法可为PTV的早期诊断及定量分析提供技术手段. |
| 关 键 词： | 猪捷申病毒 实时定量RT-PCR TaqMan |
Development of TaqMan real-time RT-PCR assay for detection of porcine teschovirus |
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| HU Feng , ZHAO Ya-rong , LV Chao-chao , WANG Gui-hua , CUI Yu-dong , WANG Chun-ren , CUI Shang-jin.Development of TaqMan real-time RT-PCR assay for detection of porcine teschovirus[J].Chinese Journal of Preventive Veterinary Medicine,2012,34(3):206-210. | |
| Authors: | HU Feng ZHAO Ya-rong LV Chao-chao WANG Gui-hua CUI Yu-dong WANG Chun-ren CUI Shang-jin |
| Institution: | 1,2(1.College of Animal Science and Technology,Heilongjiang August First Land Reclamation University,Daqing 163319,China;2.Division of Swine Infections Disease,State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China;3.Dabeinong Research Center of Veterinary Medicine,Beijing 100097,China) |
| Abstract: | To detect the porcine teschovirus(PTV),a real-time RT-PCR based on TaqMan was established with a pair of primers and a TaqMan probe targeting the highly conserved sequences of the 5’-untranslated region of 1 to 11 serotypes of PTV.The results indicated that the real-time RT-PCR was specific for detection of PTV with a detection limit of 10 copies,but not for porcine parvovirus,porcine circovirus 2,porcine reproductive and respiratory syndrome virus,pseudorabies virus,and classical swine fever virus.The coefficient of variation of inter-assay and intra-assay were less than 3%.A total of 91 clinical samples were tested by the real-time RT-PCR comparing with virus isolation,and positive rates were 79.12%(72/91) and 57.14%(48/91),respectively.In conclusion,the developed real-time RT-PCR assay was an effective method for detection and quantification of PTV in organs of infected pigs. |
| Keywords: | porcine teschovirus real-time RT-PCR TaqMan |
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