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


CONCLUSIONS
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.
Chiou
Chiuan-Chian
CC
Department of Medical Biotechnology and Laboratory Science, Chang Gung University, Tao-Yuan, Taiwan.
Chen
Tai-Long
TL
Tsao
Kuo-Chien
KC
Shih
Shin-Ru
SR
Huang
Chung-Guei
CG
Huang
Ya-Ling
YL
Chang
Chung-Ming
CM
D000483:Alleles; D000818:Animals; D001483:Base Sequence; D004196:Disease Outbreaks; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics* / Q000302:isolation & purification*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors