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Brief Communications |
1 Department of Microbiology, The University of Hong Kong, Hong Kong SAR, China;
2 Department of Microbiology, Queen Mary Hospital, Hong Kong SAR, China;
3 HKU-Pasteur Research Centre, Hong Kong SAR, China;
aaddress correspondence to these authors at: Leo L.M. Poon, Department of Microbiology, University Pathology Building, Queen Mary Hospital, Pokfulam, Hong Kong SAR, China. Fax: 852 2855 1241; e-mail: llmpoon{at}hkucc.hku.hk; or J.S. Malik Peiris, Department of Microbiology, University Pathology Bldg., Queen Mary Hospital, Pokfulam, Hong Kong SAR, China. Fax: 852-2855-1241; e-mail: malik{at}hkucc.hku.hk.
Abstract
Background: Influenza A viruses are medically important viral pathogens that cause significant mortality and morbidity throughout the world. The recent emergence of a novel human influenza A virus (H1N1) poses a serious health threat. Molecular tests for rapid detection of this virus are urgently needed.
Methods: We developed a conventional 1-step RT-PCR assay and a 1-step quantitative real-time RT-PCR assay to detect the novel H1N1 virus, but not the seasonal H1N1 viruses. We also developed an additional real-time RT-PCR that can discriminate the novel H1N1 from other swine and human H1 subtype viruses.
Results: All of the assays had detection limits for the positive control in the range of 1.0 x 10–4 to 2.0 x 10–3 of the median tissue culture infective dose. Assay specificities were high, and for the conventional and real-time assays, all negative control samples were negative, including 7 human seasonal H1N1 viruses, 1 human H2N2 virus, 2 human seasonal H3N2 viruses, 1 human H5N1 virus, 7 avian influenza viruses (HA subtypes 4, 5, 7, 8, 9, and 10), and 48 nasopharyngeal aspirates (NPAs) from patients with noninfluenza respiratory diseases; for the assay that discriminates the novel H1N1 from other swine and human H1 subtype viruses, all negative controls were also negative, including 20 control NPAs, 2 seasonal human H1N1 viruses, 2 seasonal human H3N2 viruses, and 2 human H5N1 viruses.
Conclusions: These assays appear useful for the rapid diagnosis of cases with the novel H1N1 virus, thereby allowing better pandemic preparedness.
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