| H1N1和H3N2亚型流感病毒基因芯片检测方法的建立 |
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| 引用本文： | 王慧煜,梅琳,侯义宏,李全芬,林祥梅,韩雪清.H1N1和H3N2亚型流感病毒基因芯片检测方法的建立[J].中国兽医科学,2011(12). |
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| 作者姓名： | 王慧煜 梅琳 侯义宏 李全芬 林祥梅 韩雪清 |
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| 作者单位： | 中国检验检疫科学研究院; |
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| 基金项目： | 质检公益性行业科研专项(200910132); 基本科研业务费专项资金资助项目(2009JK022); 国家重点基础研究发展计划项目(2011CB504704-X) |
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| 摘 要： | 为建立同时能鉴别甲型H1N1和猪流感病毒常见亚型的新型基因芯片检测方法,根据GenBank中已发表的甲型流感病毒MP的基因序列和甲型H1N1(2009)和猪流感病毒H1N1、H3N2亚型的基因序列,设计、筛选并合成7对特异性引物和1对通用引物;根据扩增的靶序列,设计并合成14条特异性探针和3条质控探针,制备了甲型H1N1(2009)流感病毒和猪流感病毒H1N1、H3N2亚型基因芯片;并进行了特异性试验、敏感性试验和田间样品的检测。结果显示,该芯片检测方法与猪细小病毒(PPV)、猪瘟病毒(CSFV)、猪繁殖与呼吸综合征病毒(PRRSV)等猪常见病毒无交叉反应;对猪H1N1、猪H3N2和甲型H1N1(2009)流感病毒而言,最低可检测到105、104和105稀释的病毒株。结果证实,该方法特异性强、敏感性高,是一种高通量的甲型H1N1和猪流感常见亚型筛查方法。
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| 关 键 词： | 甲型H1N1 猪流感病毒 基因芯片 检测 |
Establishment of oligonucleotide microarray for detection of influenza virus subtypes H1N1 and H3N2 |
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| WANG Hui-yu,MEI Lin,HOU Yi-hong,LI Quan-fen,LIN Xiang-mei,HAN Xue-qing.Establishment of oligonucleotide microarray for detection of influenza virus subtypes H1N1 and H3N2[J].Veterinary Science in China,2011(12). |
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| Authors: | WANG Hui-yu MEI Lin HOU Yi-hong LI Quan-fen LIN Xiang-mei HAN Xue-qing |
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| Institution: | WANG Hui-yu,MEI Lin,HOU Yi-hong,LI Quan-fen,LIN Xiang-mei,HAN Xue-qing(Chinese Academy of Inspection and Quarantine,Beijing 100029,China) |
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| Abstract: | Seven pairs of primers specific for different subtypes and a pair of universal primers were carefully designed based on the genomic sequences of A/H1N1 and swine influenza virus retrieved from GenBank database.Several multiplex RT-PCR methods were then developed.Further 14 oligonucleotide probes specific for A/H1N1 and swine influenza virus were designed according to the published gene in target cDNA domains.Then a microarray for A/H1N1 and swine influenza virus was developed with its specificity and sensit... |
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| Keywords: | A/H1N1 swine influenza virus microarray detection |
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