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Establishment of a MGB TaqMan Probe-based Real-time Quantitative PCR Detection Method for Duck (Anas platyrhynchos domestica) Adenovirus Type 3 |
CHEN Cui-Teng, CHEN Zhen, ZHU Chun-Hua, CAI Guo-Zhang, LIU Bing-Qiong, HUANG Yu* |
Institute of Animal Husbandry & Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou 350013, China |
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Abstract Duck (Anas platyrhynchos domestica) adenovirus type 3 (DAdV-3) is a newly discovered duck-derived adenovirus in recent years. In this study, specific primers and probe were designed based on the 22K gene characteristics of DAdV-3 representative strain (CH-GD-12-2014 strain, GenBank No. KR135164) in the NCBI database, and a MGB TaqMan probe real-time quantitative PCR method for detecting DAdV-3 was established. The results showed that the established qRT-PCR method had the following characteristics: High sensitivity, minimum detection limit of 55 copies/μL; Strong specificity, and no cross-reaction with common infectious disease pathogens in duck flocks; Good reproducibility, intra-group and inter-group repetitive experimental variation coefficients were 0.44%~2.10% and 0.64%~2.37%, respectively. 61 duck-derived materials were detected by the established method, and 15 positive DAdV-3 samples were detected with a positive rate of 24.6%. This study laid the foundation for rapid laboratory diagnosis and epidemiological investigation of DAdV-3 infection.
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Received: 20 August 2018
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Corresponding Authors:
* huangyu_815@163.com
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