Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 163, Issue 2, Pages 495-497Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2009.10.025
Keywords
Influenza; H1N1; Real-time; Molecular beacon; Nucleic acid sequence-based amplification (NASBA)
Funding
- Important National Science & Technology Specific Projects [2008ZX10004-002]
- National Natural Science Foundation of China [30901285]
- Natural Science Foundation of Jiangsu Province [SBK200922783]
- Jiangsu Provincial International Cooperation Program [BE2008683]
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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). (C) 2009 Elsevier B.V. All rights reserved.
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