Cheng-Dong Yu, Yu-Ri Choi, Jong-Yeol Park, Sang-Won Kim, Se-Yeoun Cha, Hyung-Kwan Jang, Min Kang, Bai Wei
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Establishment and Application of Mismatch Amplification Mutation Assay-PCR for Rapid Detection and Differentiation of Duck Hepatitis A Virus-1 Attenuated Vaccine and Wild Strains.
Duck hepatitis A virus type 1 (DHAV-1) is the main pathogen causing viral hepatitis in ducks, marked by high contagion and acute mortality. Live attenuated DHAV-1 vaccines are widely used to control the disease. This study aims to develop a mismatch amplification mutation assay (MAMA)-PCR for the rapid detection and differentiation of Korean DHAV-1 wild-type strains from vaccine strains. A MAMA primer was designed to target a single nucleotide polymorphism (SNPs) at position 2276 within the VP1 gene, allowing differentiation in a single PCR reaction. The MAMA-PCR accurately identified both strains, with detection limits of 100.5 ELD50/mL and 102.3 ELD50/mL, respectively. The MAMA-PCR demonstrated specificity, showing no cross-reactivity with 12 other viral and bacterial pathogens. The MAMA-PCR was applied to 89 farms, yielding results consistent with nested-PCR and sequence determination, identifying four positive farms for DHAV-1 vaccine strains. In conclusion, this study is the first to employ the MAMA-PCR method to distinguish between DHAV-1 wild-type and vaccine strains. The developed method is rapid, simple, specific, and sensitive, thereby serving as an effective tool for clinical diagnostics in identifying and differentiating between Korean DHAV-1 wild-type and vaccine strains.
AnimalsAgricultural and Biological Sciences-Animal Science and Zoology
CiteScore
4.90
自引率
16.70%
发文量
3015
审稿时长
20.52 days
期刊介绍:
Animals (ISSN 2076-2615) is an international and interdisciplinary scholarly open access journal. It publishes original research articles, reviews, communications, and short notes that are relevant to any field of study that involves animals, including zoology, ethnozoology, animal science, animal ethics and animal welfare. However, preference will be given to those articles that provide an understanding of animals within a larger context (i.e., the animals'' interactions with the outside world, including humans). There is no restriction on the length of the papers. Our aim is to encourage scientists to publish their experimental and theoretical research in as much detail as possible. Full experimental details and/or method of study, must be provided for research articles. Articles submitted that involve subjecting animals to unnecessary pain or suffering will not be accepted, and all articles must be submitted with the necessary ethical approval (please refer to the Ethical Guidelines for more information).