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.
[1] 周镇海. 2016. 鸭腺病毒3型基因组序列分析及致病性研究[D]. 硕士学位论文, 华南农业大学, 导师: 钟仰进, pp. 8-63. (Zhou Z H.2016. Genonmic sequence analysis and pathogenecity study of duck adenovirus 3[D]. Thesis for M.S., South China Agricultural University, Suppervisor: Zhong Y J, pp. 8-63.) [2] Ahi Y S, Mittal S K.2016. Components of adenovirus genome packaging[J]. Frontiers in Microbiology, 7:1503 [3] Biasiotto R, Akusjärvi G.2015. Regulation of human adenovirus alternative RNA splicing by the adenoviral L4-33K and L4-22K proteins[J]. International Journal of Molecular Sciences, 16(2): 2893-2912. [4] Bouquet J F, Moreau Y, McFerran J B, et al.1982. Isolation and characterisation of an adenovirus isolated from Muscovy ducks[J]. Avian Pathology, 11(2): 301-307. [5] Cha S Y, Kang M, Moon O K, et al.2013. Respiratory disease due to current Egg drop syndrome virus in Pekin ducks[J]. Veterinary Microbiology, 165(3-4): 305-311. [6] Das S, Fearnside K, Sarker S, et al.2017. A novel pathogenic aviadenovirus from red-bellied parrots (Poicephalus rufiventris) unveils deep recombination events among avian host lineages[J]. Virology, 502: 188-197. [7] Guimet D, Hearing P.2013. The adenovirus L4-22K protein has distinct functions in the posttranscriptional regulation of gene expression and encapsidation of the viral genome[J]. Journal of Virology, 87(13): 7688-7699. [8] Kaján G L, Stefancsik R, Ursu K, et al.2010. The first complete genome sequence of a non-chicken aviadenovirus, proposed to be turkey adenovirus 1[J]. Virus Research, 153(2): 226-233. [9] Kang M, Cha S Y, Jang H K.2017. Tropism and infectivity of duck-derived egg drop syndrome virus in chickens[J]. PLoS One, 12(5): e0177236. [10] Kutyavin I V, Afonina I A, Mills A, et al.2000. 3'-minor groove binder-DNA probes increase sequence specificity at PCR extension temperatures[J]. Nucleic Acids Research, 28(2): 655-661. [11] Lan S, Kamel W, Punga T, et al.2017. The adenovirus L4-22K protein regulates transcription and RNA splicing via a sequence-specific single-stranded RNA binding[J]. Nucleic Acids Research, 45(4): 1731-1742. [12] Li C J, Li H Y, Wang D D, et al.2016. Characterization of fowl adenoviruses isolated between 2007 and 2014 in China[J]. Veterinary Microbiology, 197(): 62-67. [13] Marek A, Kaján G L, Kosiol C, et al.2014. Complete genome sequences of pigeon adenovirus 1 and duck adenovirus 2 extend the number of species within the genus Aviadenovirus[J]. Virology, 462-463: 107-114. [14] Marek A, Ballmann M Z, Kosiol C, et al.2014. Whole-genome sequences of two turkey adenovirus types reveal the existence of two unknown lineages that merit the establishment of novel species within the genus Aviadenovirus[J]. The Journal of General Virology, 95(1): 156-170. [15] Mittal D, Jindal N, Tiwari A K, et al.2014. Characterization of fowl adenoviruses associated with hydropericardium syndrome and inclusion body hepatitis in broiler chickens[J]. Virus Disease, 25(1): 114-119. [16] Morshed R, Hosseini H, Langeroudi A G, et al.2017. Fowl adenoviruses D and E cause inclusion body hepatitis outbreaks in broiler and broiler breeder pullet flocks[J]. Avian Diseases, 61(2): 205-210. [17] Niu Y J, Sun W, Zhang G H, et al.2016. Hydropericardium syndrome outbreak caused by fowl adenovirus serotype 4 in China in 2015[J]. The Journal of General Virology, 97(10): 2684-2690. [18] Pitcovski J, Mualem M, Rei-Koren Z, et al.1998. The complete DNA sequence and genome organization of the avian adenovirus, hemorrhagic enteritis virus[J]. Virology, 249(2): 307-315. [19] Said A, Wang W, Woldermariam T, et al.2018. Domains of bovine adenovirus-3 protein 22K involved in interacting with viral protein 52K and cellular importins α-5/α-7[J]. Virology, 522: 209-219. [20] To K K, Tse H, Chan W M, et al.2014. A novel psittacine adenovirus identified during an outbreak of avian chlamydiosis and human psittacosis: Zoonosis associated with virus-bacterium coinfection in birds[J]. PLoS Neglected Tropical Diseases, 8(12): e3318. [21] Uchiyama M, Maesawa C, Yashima-Abo A, et al.2005. Consensus JH gene probes with conjugated 3'-minor groove binder for monitoring minimal residual disease in acute lymphoblastic leukemia[J]. The Journal of Molecular Diagnostics, 7(1): 121-126. [22] Van Eck J H, Davelaar F G, Heuvel-Plesman T A, et al.1976. Dropped egg production, soft shelled and shell-less eggs associated with appearance of precipitins to adenovirus in flocks of laying fowls[J]. Avian Pathology, 5(4): 261-272. [23] Wang X, Qi X, Yang B, et al.2018. Autophagy benefits the replication of egg drop syndrome virus in duck embryo fibroblasts[J]. Frontiers in Microbiology, 9: 1091 [24] Wu K, Orozco D, Hearing P.2012. The adenovirus L4-22K protein is multifunctional and is an integral component of crucial aspects of infection[J]. Journal of Virology, 86(19): 10474-10483. [25] Zhang X, Zhong Y, Zhou Z, et al.2016. Molecular characterization, phylogeny analysis and pathogenicity of a Muscovy duck adenovirus strain isolated in China in 2014[J]. Virology, 493: 12-21.
