Detection of highly pathogenic zoonotic influenza virus H5N6 by reverse-transcriptase quantitative polymerase chain reaction.
Detection of highly pathogenic zoonotic influenza virus H5N6 by reverse-transcriptase quantitative polymerase chain reaction.
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DOI：
10.1186/s12985-015-0250-3
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发表时间：
2015-02-08
期刊：
影响因子：
4.8
通讯作者：
Meehan B
中科院分区：
文献类型：
--
作者：
Heine HG;Foord AJ;Wang J;Valdeter S;Walker S;Morrissy C;Wong FY;Meehan B
由于H5血凝素基因与神经氨酸酶基因NA1、Na2、Na5、Na6和Na8的重组，最近出现了高致病性H5变异型禽流感病毒。这些病毒构成新建议的HA分支2.3.4.4(以前暂时称为分支2.3.4.6)，并已在中国、韩国、老挝、日本和越南的家禽中爆发疾病，并在中国导致一人死亡。人们真正担心的是，这一新的分支可能会广泛传播，并不容易使用现有的诊断算法来识别。建立了基于荧光探针的逆转录酶定量聚合酶链式反应(RT-qPCR)方法，用于鉴定亚洲新出现的H5N6亚型新分支病毒2.3.4.4。用血凝素(HA)基因鉴定分支，用NA基因鉴定N6。使用小型凹槽粘合剂(MGB)探针的HA检测能够检测和区分属于拟议分支2.3.4.4的A/鸭子/LAOS/XBY004/2014(H5N6)和相关的甲型流感(H5N6)病毒分离株与其他H5 HPAI病毒。此外，欧亚N6试验能够将N6与其他NA亚型区分开来。代表拟议分支2.3.4.4的老挝甲型流感病毒(H5N6)通过分支特异的HA RT-qPCR方法检测到并与其他H5N1分支中的病毒区分开来，而N6-NA亚型则通过欧亚N6 RT-qPCR方法确定。这种针对支系的化验对于监测和诊断实验室来说具有特别的价值，因为这些实验室还不能轻易获得测序。
Variant high pathogenicity avian influenza (HPAI) H5 viruses have recently emerged as a result of reassortment of the H5 haemagglutinin (HA) gene with different neuraminidase (NA) genes, including NA1, NA2, NA5, NA6 and NA8. These viruses form a newly proposed HA clade 2.3.4.4 (previously provisionally referred to as clade 2.3.4.6), and have been implicated in disease outbreaks in poultry in China, South Korea, Laos, Japan and Vietnam and a human fatality in China. There is real concern that this new clade may be wide spread and not readily identified using existing diagnostic algorithms. Fluorescent probe based reverse-transcriptase quantitative polymerase chain reaction (RT-qPCR) assays were developed to facilitate the identification of novel clade 2.3.4.4 viruses of H5N6 subtype emerging in Asia. Assays were aimed at the haemagglutinin (HA) gene for clade identification and at the NA gene to identify N6. The HA assay employing a minor groove binder (MGB) probe was able to detect and differentiate A/duck/Laos/XBY004/2014(H5N6) and related influenza A(H5N6) virus isolates belonging to the proposed clade 2.3.4.4 from other H5 HPAI viruses. In addition, an Eurasian N6 assay was able to differentiate N6 from other NA subtypes. Laos influenza A(H5N6) virus representative of proposed clade 2.3.4.4, was detected and differentiated from viruses in other H5N1 clades using a clade-specific HA RT-qPCR assay whereas the N6-NA subtype was determined by an Eurasian N6 RT-qPCR assay. Such a clade-specific assay would be of particular value for surveillance and in diagnostic laboratories where sequencing is not readily available.
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影响因子：
11.8
作者：
Gu, Min;Zhao, Guo;Liu, Xiufan
通讯作者：
Liu, Xiufan
影响因子：
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作者：
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通讯作者：
Ostell J
影响因子：
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作者：
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通讯作者：
LIPMAN, DJ