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H4亚型和N2亚型禽流感病毒二重RT-PCR检测方法的建立 被引量:1

Establishment of a Duplex RT-PCR Assay for Detection of H4 and N2 Subtype Avian Influenza Virus
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摘要 为建立一种简单、快速鉴别H4亚型和N2亚型禽流感病毒(AIV)的方法,针对H4亚型AIV HA基因和N2亚型AIV NA基因的保守区域,分别设计筛选出1对特异性引物,通过优化反应条件,建立了H4亚型和N2亚型AIV二重RT-PCR检测方法。该法可特异性扩增H4亚型和N2亚型AIV,对其他亚型AIV和常见禽病病原体无交叉反应;对H4亚型和N2亚型AIV的检测下限均为104拷贝/μL;对152份临床样品的检测结果与病毒分离鉴定结果一致。本研究建立的二重RT-PCR方法为H4亚型和N2亚型AIV的诊断提供了快速、特异、敏感和有效的检测方法。 In order to establish a simple and rapid method for detecting H4 and N2 subtype avian influenza virus(AIV),one pairs of specific primers were designed and screened respectively based on the conservative areas of HA and NA genes of the two subtypes of virus,then a duplex RT-PCR assay was established after optimization of reaction conditions,by which,H4 and N2 subtype AIV could be specifically amplified,while other subtypes AIV and the pathogens of common poultry diseases failed to be amplified;the detection limits for H4 and N2 subtype were all 104 copies/μL;the testing results of 152 clinical samples were consistent with that of virus isolation and identification.In short,the RT-PCR assay established was rapid,specific,sensitive and effective for detecting H4 and N2 subtype AIV.
作者 罗思思 谢芝勋 李孟 黄娇玲 李丹 谢丽基 张民秀 曾婷婷 王盛 张艳芳 范晴 谢志勤 邓显文 Luo Sisi;Xie Zhixun;Li Meng;Huang Jiaoling;Li Dan;Xie Liji;Zhang Minxiu;Zeng Tingting;Wang Sheng;Zhang Yanfang;Fan Qing;Xie Zhiqin;Deng Xianwen(Guangxi Veterinary Research Institute,Guangxi Key Laboratory of Veterinary Biotechnology,Nanning,Guangxi 530001,China)
出处 《中国动物检疫》 CAS 2020年第1期69-73,共5页 China Animal Health Inspection
基金 广西科技重大专项(桂科AA17204057) 广西科技基地和人才专项(桂科AD17195083) 桂科专项(17-1) “广西八桂学者”专项(2019.79) 国家“万人计划”领军人才专项(W02060083)
关键词 禽流感病毒 H4亚型 N2亚型 二重RT-PCR 检测方法 avian influenza virus H4 subtype N2 subtype duplex RT-PCR detection method
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  • 1陈溥言.应用酶联免疫吸附试验检测雏鸭肝炎病毒抗原[J].畜牧与兽医,1989,21(5):200-201. 被引量:12
  • 2谢芝勋,董建宝,唐小飞,刘加波,庞耀珊,邓显文,谢志勤.3株H9N2亚型禽流感病毒广西分离株全基因组序列的测定与分析[J].中国人兽共患病学报,2007,23(9):916-922. 被引量:9
  • 3Murty D K, Hanson L E.A modified microgel diffusion method and its application in the study of the virus of duck hepatitis [J]. Am J Vet Res, 1961,22(3 ): 274-278.
  • 4Kim M C, Kwon Y K, Joh S J, et al.Development of one step reverse transcriptase polymerase chain reaction to detect duck hepatitis virus type I [J].Avian Dis, 2007, 51 (2): 540-545.
  • 5Sambrook J,Fritsh E T, Maniatis T.Molecular Cloning: a laboratory manuel [M].New York: Cold Spring Harbor Laboratory Press, 1989.
  • 6Xie Z,Xie L,Fan Q,et al. A duplex quantitative real-time PCR assay for the detection of Haplosporidium and Perkinsus spe- cies in Shellfish[J]. ParasitoI Res,2013,112 (4):1597-1606.
  • 7Xie Z,Pang Y S,Liu J,et al. A multiplex RT-PCR for detec- tion of type a influenza virus and differentiation of avian HS, H7 and H9 hemagglutinin subtype[J]. Mol Cell Probes, 2006,20(3-4) : 245-249.
  • 8Peng Y,Xie Z,Liu J,et al. Visual detection of H3 subtybe avi- an influenza viruses by reverse transcription loop-mediated iso- thermal amplifieation assay[J]. Virol J, 2011,8 : 337.
  • 9Nishiura H, Mizumoto K, Ejima K. How to interpret the transmissibility of novel influenza A(HTN9) : an analysis of initial epidemiological data of human cases from China[J]. 2013,10:30.
  • 10Liu D, Shi W, Shi Y, et al. Origin and diversity of novel avian influenza A HTN9 viruses causing human infection: phyloge- netic, structural, and coalescent analyses[J]. Lancet, 2013, 381 (9881) : 1926-1932.

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