Development of real-time RT-PCR for the detection of avian influenza virus
Development of real-time RT-PCR for the detection of avian influenza virus
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DOI：
10.1637/0005-2086-47.s3.1079
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发表时间：
2003-01-01
期刊：
影响因子：
1.4
通讯作者：
Suarez, DL
中科院分区：
文献类型：
--
作者：
Spackman, E;Senne, DA;Suarez, DL
采用水解探针建立了实时逆转录酶/聚合酶链反应（RRT-PCR）检测禽流感病毒（AIV）及其H5和H7亚型的方法。AIV特异性引物和探针被定向到大多数A型流感病毒中保守的AIV基质基因区域。H5和H7引物和探针指向北美禽流感病毒中保守的H5和H7血凝素基因区域。将该RRT-PCR检测方法的灵敏度和特异性与从纽约和新泽西109个活禽市场（LBMs）采集的1550份临床拭子样本的鸡胚病毒分离（VI）进行比较。65个流感病毒阳性样本来源的lbm中有61个（93.8%）检测到流感病毒。在58个血凝抑制试验H7流感阳性的市场中，RRT-PCR检测到H7流感的市场有56个（96.5%）。本研究中H5阳性样本太少，无法验证H5 RRT-PCR检测。虽然RRT-PCR对单个样本的敏感性低于VI，但本研究表明，AIV和H7 RRT-PCR检测是快速筛选禽群和lbm的良好工具。
A real-time reverse transcriptase/polymerase chain reaction (RRT-PCR) assay was developed using hydrolysis probes for the detection of avian influenza virus (AIV) and the H5 and H7 subtypes. The AIV specific primers and probes were directed to regions of the AIV matrix gene that are conserved among most type A influenza viruses. The H5 and H7 primers and probes are directed to H5 and H7 hemagglutinin gene regions that are conserved among North American avian influenza viruses. The sensitivity and specificity of this RRT-PCR assay was compared to virus isolation (VI) in chicken embryos with 1550 clinical swab samples from 109 live-bird markets (LBMs) in New York and New Jersey. RRT-PCR detected influenza in samples from 61 of 65 (93.8%) of the LBMs that were the sources of VI positive samples. Of the 58 markets that were positive for H7 influenza by hemagglutination inhibition assay, RRT-PCR detected H7 influenza in 56 markets (96.5%). Too few H5 positive samples were obtained to validate the H5 RRT-PCR assay in this study. Although RRT-PCR was less sensitive than VI on an individual sample basis, this study demonstrated that the AIV and H7 RRT-PCR assays are good tools for the rapid screening of flocks and LBMs.