Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2009 Aug;47(8):2675-7.
doi: 10.1128/JCM.01087-09. Epub 2009 Jun 24.

Detection of novel (swine origin) H1N1 influenza A virus by quantitative real-time reverse transcription-PCR

Ruixue Wang, Zong-Mei Sheng, Jeffery K Taubenberger

Detection of novel (swine origin) H1N1 influenza A virus by quantitative real-time reverse transcription-PCR

Ruixue Wang et al. J Clin Microbiol. 2009 Aug.

Erratum in

  • J Clin Microbiol. 2009 Oct;47(10):3403
No abstract available

PubMed Disclaimer

Figures

FIG. 1.
FIG. 1.
RT-PCR for H1 subtype HAs. (A) RT-PCR amplification with the universal H1 HA primer set (set 3). cDNAs from swine, human, and avian origin H1 subtype influenza A viruses were amplified. The final reaction mixture contained 1× PCR Gold buffer, 2.5 mM MgCl2, 0.2 mM deoxynucleoside triphosphate mixture, 4 U AmpliTaq Gold polymerase (Applied Biosystems, Foster City, CA), and 0.6 μM primers. Thermocycling was performed in a DNAEngine (Bio-Rad, Hercules, CA) under the following cycling conditions: 10 min at 95°C, followed by 40 cycles of 94°C for 30 s, 60°C for 30 s, and 72°C for 1 min. PCR products were electrophoresed on a 2% Tris-acetate-EDTA agarose gel, stained with ethidium bromide, and photographed under UV transillumination. Lanes (left to right): 1, 100-bp DNA molecular weight ladder; 2, A/swine/Ohio/23/1935(H1N1), 3, A/swine/Jamesburg/1942 (H1N1), 4, A/swine/Wisconsin/1/1967 (H1N1); 5, A/Maryland/NIH-37/2009 (H1N1); 6, A/Maryland/NIH-39/2009 (H1N1); 7, A/mallard/Ohio/171/1990 (H1N1); 8, A/green-winged teal/Ohio/72/1999(H1N1); 9, A/green-winged teal/Ohio/430/1987 (H1N1); 10, A/California/04/2009 (H1N1); 11, A/New York/470/2004 (H3N2); 12, water. (B) Amplification curve for influenza A virus H1 HA real-time RT-PCR assay. Shown are the 10−1 to 10−5 dilutions of A/California/04/2009 (H1N1) cDNA (final concentrations, 0.4 ng to 40 fg viral RNA per reaction mixture) with the FAM (novel swine origin) probe. Reactions were run in duplicate. The final reaction mixture contained 1× ABI master mix (Applied Biosystems), 0.9 μM primers, and 0.25 μM of each probe under the following cycling conditions: 2 min at 50°C and 10 min at 95°C, followed by 40 cycles of 95°C for 15 s and 60°C for 1 min.
See this image and copyright information in PMC

References

    1. Centers for Disease Control and Prevention. 12 May 2009, posting date. H1N1 Flu (Swine Flu): Resources for Laboratories. Centers for Disease Control and Prevention, Atlanta, GA. http://www.cdc.gov/h1n1flu/lab/.
    1. Garten, R. J., C. T. Davis, C. A. Russell, B. Shu, S. Lindstrom, A. Balish, W. M. Sessions, X. Xu, E. Skepner, V. Deyde, M. Okomo-Adhiambo, L. Gubareva, J. Barnes, C. B. Smith, S. L. Emery, M. J. Hillman, P. Rivailler, J. Smagala, M. de Graaf, D. F. Burke, R. A. Fouchier, C. Pappas, C. M. Alpuche-Aranda, H. Lopez-Gatell, H. Olivera, I. Lopez, C. A. Myers, D. Faix, P. J. Blair, C. Yu, K. M. Keene, P. D. Dotson, Jr., D. Boxrud, A. R. Sambol, S. H. Abid, K. St. George, T. Bannerman, A. L. Moore, D. J. Stringer, P. Blevins, G. J. Demmler-Harrison, M. Ginsberg, P. Kriner, S. Waterman, S. Smole, H. F. Guevara, E. A. Belongia, P. A. Clark, S. T. Beatrice, R. Donis, J. Katz, L. Finelli, C. B. Bridges, M. Shaw, D. B. Jernigan, T. M. Uyeki, D. J. Smith, A. I. Klimov, and N. J. Cox. 22 May 2009. Antigenic and genetic characteristics of swine-origin 2009 A(H1N1) influenza viruses circulating in humans. Science doi:10.1126/science.1176225. - DOI - PMC - PubMed
    1. Krafft, A. E., K. L. Russell, A. W. Hawksworth, S. McCall, M. Irvine, L. T. Daum, J. L. Connoly, A. H. Reid, J. C. Gaydos, and J. K. Taubenberger. 2005. Evaluation of PCR testing of ethanol-fixed nasal swab specimens as an augmented surveillance strategy for influenza virus and adenovirus identification. J. Clin. Microbiol. 431768-1775. - PMC - PubMed
    1. Layne, S. P., A. S. Monto, and J. K. Taubenberger. 2009. Pandemic influenza: an inconvenient mutation. Science 3231560-1561. - PMC - PubMed
    1. Poon, L. L., K. H. Chan, G. J. Smith, C. S. Leung, Y. Guan, K. Y. Yuen, and J. S. Peiris. 21 May 2009. Molecular detection of a novel human influenza (H1N1) of pandemic potential by conventional and real-time quantitative RT-PCR assays. Clin. Chem. doi:10.1373/clinchem.2009.130229. - DOI - PMC - PubMed

Publication types

MeSH terms