Uracil-DNA glycosylase-treated reverse transcription loop-mediated isothermal amplification for rapid detection of avian influenza virus preventing carry-over contamination.
10.4142/jvs.2016.17.3.421
- Author:
Eun Mi KIM
1
;
Hyo Sung JEON
;
Ji Jung KIM
;
Yeun Kyung SHIN
;
Youn Jeong LEE
;
Sang Geon YEO
;
Choi Kyu PARK
Author Information
1. College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University, Daegu 41566, Korea. parkck@knu.ac.kr
- Publication Type:Brief Communication
- Keywords:
avian influenza virus;
contamination;
loop-mediated isothermal amplification;
uracil-DNA glycosylase
- MeSH:
Animals;
Deoxyuridine;
Diagnosis;
DNA;
Influenza in Birds*;
Limit of Detection;
Reverse Transcription*;
Uracil-DNA Glycosidase
- From:Journal of Veterinary Science
2016;17(3):421-425
- CountryRepublic of Korea
- Language:English
-
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.
