Detection of novel swine origin influenza A virus (H1N1) by real-time nucleic acid sequence-based amplification.
Rapid detection of novel swine origin influenza A virus (S-OIV) (H1N1) is crucial for timely implementation of infection control measures. In this study, a haemagglutinin (HA) gene-based real-time nucleic acid sequence-based amplification (NASBA) assay was developed for the specific detection of S-OIV (H1N1). The assay was evaluated and validated by comparing it with existing detection methods for S-OIV (H1N1). Results obtained in a 10-fold dilution series assay demonstrated the analytic sensitivity of the present assay was comparable to that of a commercial S-OIV (H1N1) real-time RT-PCR kit and higher than that of the Centers for Disease Control and Prevention (CDC) TaqMan assay. The actual detection limit of the real-time NASBA assay was approximately 50 copies per reaction. Compared with reference methods (viral culture, conventional RT-PCR, and real-time RT-PCR), the sensitivity, specificity, positive predictive value, and negative predictive value of the present assay were all 100%. Overall, the results showed that the real-time NASBA assay could be used for sensitive and specific detection of S-OIV (H1N1).
Ge
Yiyue
Y
Institute of Microbiology, Jiangsu Provincial Center for Diseases Prevention and Control, 172 Jiangsu Road, Nanjing 210009, China.
Cui
Lunbiao
L
Qi
Xian
X
Shan
Jun
J
Shan
Yunfeng
Y
Qi
Yuhua
Y
Wu
Bing
B
Wang
Hua
H
Shi
Zhiyang
Z
D006389:Hemagglutinins, Viral / Q000235:genetics; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000302:isolation & purification*; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology; D011237:Predictive Value of Tests; D021161:Self-Sustained Sequence Replication / Q000379:methods*; D012680:Sensitivity and Specificity
D006389:Hemagglutinins, Viral; C458881:hemagglutinin fusogenic peptide, influenza virus