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KMID : 0893420160170030421
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Journal of Veterinary Science
2016 Volume.17 No. 3 p.421 ~ p.425
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Uracil-DNA glycosylase-treated reverse transcription loop-mediated isothermal amplification for rapid detection of avian influenza virus preventing carry-over contamination
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Kim Eun-Mi
Jeon Hyo-Sung
Kim Ji-Jung
Shin Yeun-Kyung
Lee Youn-Jeong
Yeo Sang-Geon
Park Choi-Kyu
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Abstract
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Here, we describe a uracil-DNA glycosylase (UNG)-treated reverse transcription loop-mediated isothermal amplification (uRT-LAMP) for the visual detection of all subtypes of avian influenza A virus (AIV). The uRT-LAMP assay can prevent unwanted amplification by carryover contamination of the previously amplified DNA, although the detection limit of the uRT-LAMP assay is 10-fold lower than that of the RT-LAMP without a UNG treatment. To the best of our knowledge, this is the first successful application of deoxyuridine triphosphate/UNG strategy in RT-LAMP for AIV detection, and the assay can be applied for the rapid, and reliable diagnosis of AIVs, even in contaminated samples.
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KEYWORD
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avian influenza virus, contamination, loop-mediated isothermal amplification, uracil-DNA glycosylase
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