title: Rapid detection and differentiation of swine-origin influenza A virus (H1N1/2009) from other seasonal influenza A viruses.
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&#160;the&#160;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.
authors.affiliation: Laboratory of Molecular Virology, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892, USA. jiangqin.zhao@fda.hhs.gov
authors.forename: Jiangqin
authors.initials: J
authors.lastname: Zhao
authors.forename: Xue
authors.initials: X
authors.lastname: Wang
authors.forename: Viswanath
authors.initials: V
authors.lastname: Ragupathy
authors.forename: Panhe
authors.initials: P
authors.lastname: Zhang
authors.forename: Wei
authors.initials: W
authors.lastname: Tang
authors.forename: Zhiping
authors.initials: Z
authors.lastname: Ye
authors.forename: Maryna
authors.initials: M
authors.lastname: Eichelberger
authors.forename: Indira
authors.initials: I
authors.lastname: Hewlett
mesh_terms: D000818:Animals; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics*; D009980:Influenza A virus / Q000145:classification / Q000235:genetics*; D058889:Molecular Typing / Q000379:methods; D009439:Neuraminidase / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods
chemical_list: D019267:Hemagglutinin Glycoproteins, Influenza Virus; D009439:Neuraminidase