Detection of pandemic (H1N1) 2009 influenza virus by allele discrimination

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• 作者：Chiuan-Chian Chiou ; Tai-Long Chen ; Kuo-Chien Tsao ; Shin-Ru Shih ; Chung-Guei Huang ; Ya-Ling Huang ; Chung-Ming Chang
• 关键词：Influenza virus ; Pandemic ; Allele discrimination ; Real-time RT-PCR
• 刊名：Clinica Chimica Acta
• 出版年：2010
• 出版时间：5 August 2010
• 年：2010
• 卷：411
• 期：15-16
• 页码：1080-1083
• 全文大小：399 K

文摘

Background

The pandemic (H1N1) 2009 has become a threat of public health. To manage rapidly increased infections and disease control, a simple and reliable first-line screening test for viral infection is on urgent demand.

Methods

Through comprehensive bioinformatics analysis, a single nucleotide polymorphism in nucleoprotein gene which differentiates swine lineage virus and human seasonal H1N1 virus was selected as target. A TaqMan probe-based allele discrimination analysis was designed to analyze clinical samples. In total, 93 clinical specimens and 39 viral isolates were used to test the assay efficacy. Traditional viral culture and molecular analysis were used as gold standard.

Results

The testing results showed that the established assay has high sensitivity and specificity (92 % and 100 % ) for pandemic (H1N1) 2009. The assay could detect as low as 5 copies of NP gene of pandemic (H1N1) 2009 or 2 viral particles of human seasonal H1N1.

Conclusion

This assay can be used as a first-line screening and confirmation test for pandemic (H1N1) 2009 virus and human seasonal flu in one-tube reaction. The assay can serve as a convenient method to reduce the burden of PCR manipulation for diagnostic laboratories when large amount of samples need to be analyzed in a short time.