Journal of Food, Agriculture and Environment
Vol 10,
Issue 2,2012
Online ISSN: 1459-0263
Print ISSN: 1459-0255
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Development and application of a rapid detection system for swine influenza virus by a reverse transcription loop-mediated isothermal amplification assay
Author(s):
Ye-bing Liu 1*,
Jin-qing Jiang 2, Lei Zhang 1, Lei Chen 1, Yi-bao Ning 1Recieved Date: 2012-01-11, Accepted Date: 2012-05-02
Abstract:Based on the published sequences of different serotypes of swine influenza virus (SIV), six degenerate primers were designed to target the conserved region of the SIV polymerase A (PA) gene. An SIV reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was established by employing the genomic material from SIV H1, H3, H5, and H9 subtype strains as templates. Reactions performed using a Loopamp LA-320 turbidimeter demonstrated that the four SIV subtype strains were specifically amplified within 45 min under isothermal conditions at 63°C. This study demonstrated that there was effective amplification of the RNA template diluted 104-fold, and 19 clinical samples were SIV positive in concordance with detection by conventional virus isolation techniques. Further validation by gene chip analysis showed that there was 100% parity compared with RT-LAMP. Of the 79 clinical samples analyzed by RT-LAMP, indicating that the assay was suitable for SIV detection and was rapid, specific, and sensitive.
Keywords:Swine influenza virus,
degenerate primer, RT-LAMPJournal: Journal of Food, Agriculture and Environment
Year: 2012
Volume: 10
Issue: 2
Category: Food and Health
Pages: 196-200
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