Abstract: We previously developed a rapid and simple gold nanoparticle(NP)-based genomic microarray assay for identification of the avian H5N1 virus and its discrimination from other influenza A virus strains (H1N1, H3N2). In this study, we expanded the platform to detect the 2009 swine-origin influenza A virus (H1N1/2009). Multiple specific capture and intermediate oligonucleotides were designed for the matrix (M), hemagglutinin (HA), and neuraminidase (NA) genes of the H1N1/2009 virus. The H1N1/2009 microarrays were printed in the same format as those of the seasonal influenza H1N1 and H3N2 for the HA, NA, and M genes. Viral RNA was tested using capture-target-intermediate oligonucleotide hybridization and gold NP-mediated silver staining. The signal from the 4 capture-target-intermediates of the HA and NA genes was specific for H1N1/2009 virus and showed no cross hybridization with viral RNA from other influenza strains H1N1, H3N2, and H5N1. All of the 3 M gene captures showed strong affinity with H1N1/2009 viral RNA, with 2 out of the 3 M gene captures showing cross hybridization with the H1N1, H3N2, and H5N1 samples tested. The current assay was able to detect H1N1/2009 and distinguish it from other influenza A viruses. This new method may be useful for simultaneous detection and subtyping of influenza A viruses and can be rapidly modified to detect other emerging influenza strains in public health settings.
Keywords: nanoparticle; H5N1; swine influenza A virus; nanomicroarray
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Zhao, J.; Wang, X.; Ragupathy, V.; Zhang, P.; Tang, W.; Ye, Z.; Eichelberger, M.; Hewlett, I. Rapid Detection and Differentiation of Swine-Origin Influenza A Virus (H1N1/2009) from Other Seasonal Influenza A Viruses. Viruses 2012, 4, 3012-3019.
Zhao J, Wang X, Ragupathy V, Zhang P, Tang W, Ye Z, Eichelberger M, Hewlett I. Rapid Detection and Differentiation of Swine-Origin Influenza A Virus (H1N1/2009) from Other Seasonal Influenza A Viruses. Viruses. 2012; 4(11):3012-3019.
Zhao, Jiangqin; Wang, Xue; Ragupathy, Viswanath; Zhang, Panhe; Tang, Wei; Ye, Zhiping; Eichelberger, Maryna; Hewlett, Indira. 2012. "Rapid Detection and Differentiation of Swine-Origin Influenza A Virus (H1N1/2009) from Other Seasonal Influenza A Viruses." Viruses 4, no. 11: 3012-3019.