| 禽呼肠孤病毒与禽白血病病毒双重RT-PCR检测方法的建立及应用 |
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| 引用本文： | 马超英.禽呼肠孤病毒与禽白血病病毒双重RT-PCR检测方法的建立及应用[J].中国畜牧兽医,2013,40(2):53-56. |
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| 作者姓名： | 马超英 |
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| 作者单位： | 青海省海西州乌兰县铜普兽医站,青海海西 817000 |
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| 摘 要： | 根椐GenBank中禽呼肠孤病毒(ARV)、禽白血病病毒(ALV)基因序列,设计2对引物,在建立鉴别各病毒单项RT-PCR技术的基础上,优化双重RT-PCR反应条件,建立2种病毒的双重RT-PCR。对同一样品中的ARV、ALV核酸模板进行双重RT-PCR扩增,结果可同时扩增ARV 485 bp、ALV 673 bp的特异性片段,而对其他5种禽病病原的PCR扩增结果均为阴性。敏感性试验结果表明,该双重RT-PCR技术能检出10 pg的ALV和10 pg的ARV模板。用31份临床病料对本研究双重RT-PCR技术和单项RT-PCR技术进行对比验证,结果显示,两者的总符合率为100%。结果表明建立的双重RT-PCR检测方法,具有特异、快速、准确的特点,可用于对这2种病毒的同时检测和鉴别诊断。
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| 关 键 词： | 双重聚合酶链反应 禽呼肠孤病毒 禽白血病病毒 |
| 收稿时间： | 2012-06-15 |
Establishment and Application of a Double RT-PCR for ARV and ALV |
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| MA Chao-ying.Establishment and Application of a Double RT-PCR for ARV and ALV[J].China Animal Husbandry & Veterinary Medicine,2013,40(2):53-56. |
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| Authors: | MA Chao-ying |
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| Institution: | Ulan County, Haixi Tongpu Town Veterinary Station, Qinghai Province, Haixi 817000, China |
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| Abstract: | To identify and differentiate rapidly the cause of clinical diseases, a double RT-PCR was developed. Two sets of specific primers were designed according to the sequences of avian reovirus (ARV) and avian leukosis virus (ALV) in the GenBank. A double polymerase chain reaction was optimized to simultaneously detect two pathogens of ARV and ALV in this article. The double RT-PCR system would amplify a 485 bp fragment for ARV and a 673 bp for ALV simultaneously or separately in the samples,depending on its infection status. But not specific band amplified from other six avian pathogenic viruses.As little as 10 pg of ARV and 10 pg of ALV RNA were detected using gel electrophoresis after double RT-PCR. To evaluate the double RT-PCR, 31 clinical samples were comparatively detected.The data showed that the double RT-PCR method was 100% coincidence with the single RT-PCR, and it could be used for detection and differential diagnosis of these two viruses. |
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| Keywords: | double RT-PCR avian reovirus avian leukosis virus |
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