| 禽流感病毒RT-PCR ELISA诊断方法的建立 |
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| 引用本文： | 杨少华,胡北侠,许传田,颜士敢,张琳,黄艳艳,张秀美.禽流感病毒RT-PCR ELISA诊断方法的建立[J].中国农学通报,2012,28(2):48-52. |
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| 作者姓名： | 杨少华 胡北侠 许传田 颜士敢 张琳 黄艳艳 张秀美 |
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| 作者单位： | 山东省农业科学院畜牧兽医研究所,山东济南,250100;山东省畜禽疫病防治与繁育重点实验室,山东济南250100 |
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| 基金项目： | 山东省自然科学基金“检测禽流感病毒的“RT-PCR.微孔板杂交.ELISA”试剂盒的研究”（Y2008D54）：现代农业产业技术体系建设专项. |
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| 摘 要： | 针对A型禽流感（AI）保守的M基因设计引物，并分别用地高辛和生物素标记，建立检测A型AIV的RT-PCRELISA方法。试验主要对RT-PCRELISA的各种反应条件进行了优化，确定了2mg／mL的链霉亲和素包被、1：3000抗地高辛的过氧化物酶及孵育时间等最适反应条件。该方法的敏感性比常规琼脂糖凝胶电泳检测方法高100倍以上，而且特异性强，克服了组织样品中AIV含量少而难以检测的困难，为禽流感的早期诊断和分子流行病学调查提供了一条新途径。
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| 关 键 词： | 禽流感病毒 RT-PCR ELlSA 诊断 |
| 收稿时间： | 2011/6/13 0:00:00 |
| 修稿时间： | 8/9/2011 12:00:00 AM |
Establishment of a RT-PCR method for detection of Avian Influenza virus |
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| Yang Shaohua
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Hu Beixia
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Xu Chuantian
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Yan Shigan
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Zhang Lin
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Huang Yanyan
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Zhang Xiumei.Establishment of a RT-PCR method for detection of Avian Influenza virus[J].Chinese Agricultural Science Bulletin,2012,28(2):48-52. |
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| Authors: | Yang Shaohua Hu Beixia Xu Chuantian Yan Shigan Zhang Lin Huang Yanyan Zhang Xiumei |
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| Institution: | 1.Institute of Animal Science and Veterinary Medicine Shandong Academy of Agricultural Science, Jinan Sandong 250100 2Shandong Key Lab of Animal Disease Control and Breeding, Jinan Sandong 250100) |
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| Abstract: | In this study a RT-PCR ELISA method was established using the primer which was designed according to M gene sequence of AIV and labeled by biotin and DIG. The optimal reaction conditions were determined including the streptavidin was 2 mg/mL and anti-digoxigenin-POD dilution titer was 1：3000. This method had a strong specificity and high sensitivity. The sensitivity was more than 100 times compared to RT-PCR, so that it could overcome the difficulty for the little AIV in the samples. It provided a new way for the molecular epidemiologic investigation and early diagnosis of AIV. |
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| Keywords: | AIV RT-PCR EL1SA diagnosis |
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