数字农科院2.0

Development of a DNA microarray-based multiplex assay of avian influenza virus subtypes H5, H7, H9, N1, and N2

文献类型： 外文期刊

作者： Shi, L.;Sun, J. -S.;Yu, X. -W.;Zheng, S. -M.;Shi, L.;Sun, J. -S.;Yang, Z. -P.;Bao, H. -M.;Jiang, Y. -P.;Xiong, Y. -Z.;Chen, H. -L.;Wang, X. -R.;Shi, L.;Sun, J. -S.;Yang, Z. -P.;Bao, H. -M.;Jiang, Y. -P.;Xiong, Y. -Z.;Chen, H. -L.;Wang, X. -R.;Shi, L.;Cao, D.;Yu, X. -W.

作者机构：

关键词： DNA microarray;avian influenza virus;subtyping

期刊名称： ACTA VIROLOGICA

ISSN： 0001-723X

年卷期： 2014 年 58 卷 1 期

页码：

收录情况： SCI

摘要： Outbreaks of highly pathogenic avian influenza have caused considerable economic losses in the poultry industry and have also resulted in human deaths since 2004. Rapid subtyping of highly pathogenic avian influenza viruses(HPAIVs) in clinical specimens is a prerequisite of prompt control of disease and prevention of its spreading. In this study, we describe development of a DNA microarray-based detection and subtyping of HPAIVs in field samples. DNA copies of matrix (M) protein genes for the H5, H7, and H9 subtypes of hemagglutinin (HA) and the N1 and N2 subtypes of neuraminidase (NA) were prepared by RT-PCR and specific primers and then spotted onto aldehyde slides to form DNA microarrays. The HPAIV samples to be tested were subjected to total RNA isolation, RT-PCR with universal primers and Cy3 labeling, and the obtained double-stranded DNAs (targets) were finally hybridized with DNA microarrays (probes). A fluorescent spot on the microarray, detected by scanning indicated positive hybridization, i.e. the involved subtype. The assay was specific as various heterologous viruses or HPAIVs of other subtypes tested were negative. No cross-hybridization among different subtypes could be detected. The assay was more sensitive than RT-PCR and chicken embryo inoculation and could be also used for field samples. Summing up, the assay has proved useful for simultaneous detection and differentiation of main epidemic HPAIV subtypes.

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