| 新型鸭呼肠孤病毒RT-PCR检测方法建立与初步应用 |
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| 引用本文： | 孔丰,袁远华,肖成谋,黄淑坚.新型鸭呼肠孤病毒RT-PCR检测方法建立与初步应用[J].广东畜牧兽医科技,2014(1):24-27. |
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| 作者姓名： | 孔丰 袁远华 肖成谋 黄淑坚 |
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| 作者单位： | [1] 遂溪县遂城畜牧兽医站,广东湛江524300 [2] 大沥镇农林服务中心,广东佛山528231 [3] 佛山科学技术学院生命科学学院,广东佛山528231 |
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| 摘 要： | 为建立一种能够快速检测新型鸭呼肠孤病毒（NDRV）的方法,本研究参考GenBank上登录的NDRV S3基因保守序列设计特异性引物.经条件优化后,建立了检测NDRV的RT-PCR方法.对其特异性、敏感性和重复性进行检验.结果显示：该方法仅能从NDRV分离毒中扩增到与预期大小相符,长度为298 bp的特异性目的片段,检测灵敏度达到83.4 pg病毒RNA;而其它病毒：番鸭呼肠孤病毒、禽呼肠孤病毒、鸭病毒性肝炎病毒、鸭瘟病毒、鸭新城疫病毒和禽流感病毒等样品的扩增结果均为阴性.采用该方法对在广东不同地区采集的15份鸭病料样品进行检测,NDRV阳性率为53.33％.表明建立的RT-PCR方法特异性强、敏感度高,可用于NDRV的临床诊断和流行病学调查.
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| 关 键 词： | 新型鸭呼肠孤病毒 检测 |
Establishment and Application of RT-PCR Method for Detection of NDRV |
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| Kong Feng,Yuan Yuanhua,Xiao Chengmou,Huang Shujian.Establishment and Application of RT-PCR Method for Detection of NDRV[J].Guangdong Journal of Animal and Veterinary Science,2014(1):24-27. |
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| Authors: | Kong Feng Yuan Yuanhua Xiao Chengmou Huang Shujian |
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| Institution: | 1.Suicheng Animal Husbandry and Veterinary Medicine Station, Suixi County, Zhanjiang 524300, China; 2.Dali Town Agricultural Service Center, Foshan 528231, China; 3.College of Life Science, Foshan University, Foshan 528231, China ) |
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| Abstract: | To develop a method for detection of new-type duck reovirus （NDRV） , a RT-PCR method was established with 1 pairs of specific primers designed based on the conserved sequences of S3 gene ofNDRV. RT-PCR results showed that a 298 bp specifical fragment could be detected only from the RNA of NDRV-QY strain, and the sensitivity of RT-PCR reached to 83.4 pg NDRV RNA. Fifteen tissue samples of sick ducks from different areas of Guangdong province were detected respectively by the RT-PCR and the positive rate was 53.33% （8/15） for NDRV which indicated that the RT-PCR method for detecting NDRV was rapid, specific and sensitive, and could be used in clinic diganoses. |
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| Keywords: | New-type duck reovirus RT-PCR detection |
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