J Vet Sci.  2016 Sep;17(3):421-425. 10.4142/jvs.2016.17.3.421.

Uracil-DNA glycosylase-treated reverse transcription loop-mediated isothermal amplification for rapid detection of avian influenza virus preventing carry-over contamination

Affiliations
  • 1College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University, Daegu 41566, Korea. parkck@knu.ac.kr
  • 2RAD Inc., Daegu 39852, Korea.
  • 3M Monitor Inc., Daegu 41914, Korea.
  • 4Virology Division, Animal and Plant Quarantine Agency, Gimcheon 39660, Korea.
  • 5Avian Disease Division, Animal and Plant Quarantine Agency, Gimcheon 39660, Korea.

Abstract

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.

Keyword

avian influenza virus; contamination; loop-mediated isothermal amplification; uracil-DNA glycosylase
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