Development and Evaluation of Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Rapid and Real-Time Detection of the Swine-Origin.

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Development and Evaluation of Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Rapid and Real-Time Detection of the Swine-Origin Influenza A H1N1 Virus Manmohan Parida, Jyoti Shukla, Shashi Sharma, Sanna Ranghia Santhosh, Vasanthapuram Ravi, Reeta Mani, Maria Thomas, Shashi Khare, Arvind Rai, Radha Kant Ratho, Sujit Pujari, Bijayanti Mishra, Putcha Venkata Lakshmana Rao, Rajagopalan Vijayaraghavan The Journal of Molecular Diagnostics Volume 13, Issue 1, Pages 100-107 (January 2011) DOI: 10.1016/j.jmoldx.2010.11.003 Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 1 Real-time kinetics of S-OIV H1 gene-specific RTLAMP showing the amplification curve. The x axis depicts the time of positivity (Tp); the y axis shows the turbidity value in terms of optical density at 400 nm. The Journal of Molecular Diagnostics 2011 13, 100-107DOI: (10.1016/j.jmoldx.2010.11.003) Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 2 A: Sensitivity of S-OIV H1 gene-specific RTLAMP assay as monitored by real-time measurement of turbidity. Shown (left to right) are the amplification curves of decreasing concentration of virus, ranging from 106 to 1 copy number of the 188 bp template (HA) gene in serial tenfold dilution. B: S-OIV H1 RTLAMP-specific standard curve as generated from the amplification plot between different copy number of the template (serial tenfold dilution from 106 to 1 copy number) and Tp. The Tp value shown here is the average of triplicates run for each concentration. The Journal of Molecular Diagnostics 2011 13, 100-107DOI: (10.1016/j.jmoldx.2010.11.003) Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 3 Quantitative determination of the virus load in terms of genome copy number in acute-phase throat-swab samples as determined from the S-OIV H RTLAMP-specific standard curve based on their Tp values. The Journal of Molecular Diagnostics 2011 13, 100-107DOI: (10.1016/j.jmoldx.2010.11.003) Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions