Detection and differentiation of four poultry diseases using asymmetric reverse transcription polymerase chain reaction in combination with oligonucleotide microarrays

文献类型： 外文期刊

第一作者： Tao, Qimeng

作者： Tao, Qimeng;Wang, Xiurong;Bao, Hongmei;Wu, Jianan;Shi, Lin;Li, Yanbing;Qiao, Chuanling;Chen, Hualan;Tao, Qimeng;Wang, Xiurong;Bao, Hongmei;Wu, Jianan;Shi, Lin;Li, Yanbing;Qiao, Chuanling;Chen, Hualan;Yakovlevich, Samuilenko Anatolij;Mikhaylovna, Poukhova Nina

作者机构：

关键词： Asymmetric reverse transcription polymerase chain reaction;Avian influenza virus;microarray;Newcastle disease virus

期刊名称：JOURNAL OF VETERINARY DIAGNOSTIC INVESTIGATION （ 影响因子：1.279； 五年影响因子：1.522 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： Asymmetric reverse transcription polymerase chain reaction (RT-PCR) and microarrays were combined to distinguish 4 viruses, including Avian influenza virus (AIV), Newcastle disease virus (NDV), Infectious bronchitis virus (IBV), and Infectious bursal disease virus (IBDV), and hemagglutinin (HA) subtypes H5, H7, and H9, and neuraminidase (NA) subtypes N1 and N2 of AIV. The AIV matrix protein (M), and HA and NA genes, IBV nucleoprotein (NP) gene, NDV NP gene, and IBDV A fragment gene were cloned into plasmids. These genes were amplified from these positive recombinant plasmids, which included the inserted target genes by PCR. The PCR products were purified and printed on the amino-modified slides as the probes. RNA was extracted from samples and labeled by asymmetric RT-PCR using a cyanine (Cy)3-labeled primers. The labeled complementary (c)DNA was hybridized to the probes immobilized on the glass slides. After hybridization, the microarrays were scanned, and the hybridization pattern agreed perfectly with the known location of each probe. No cross-hybridization could be detected. Results demonstrated that microarray based on asymmetric RT-PCR is an effective way to distinguish AIV, IBV, NDV, and IBDV simultaneously.

分类号： S85

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