猪丁型冠状病毒CH/GX/1468B/2017的分离鉴定及全基因组序列分析

doi:10.16431/j.cnki.1671-7236.2019.07.004

Abstract

Abstract:

The small intestine tissue and its contents of diarrhea piglets from porcine deltacoronavirus (PDCoV) suspected case were collected for virus isolation in the study,the isolated virus were identified by typical cytopathic effect (CPE),RT-PCR,IFA and genomic sequence analysis.The results showed that a PDCoV strain named CH/GX/1468B/2017 (PDCoV 1468B) was isolated from piglets.The strain could grow and proliferate steadily and effectively in LLC-PK cells with typical cytopathic changes.The strain has been passaged for 15 generations in LLC-PK cells.The infectious titers of the viruses gradually increasing and stabilized above 10^8.10 TCID₅₀/mL.The complete genome sequence of PDCoV 1468B was 25 399 nt in length,excluding the poly(A) tail.Genomic analysis showed that PDCoV 1468B shared 97.2% to 99.4% nucleotide identity with other 23 reference PDCoV strains (GenBank),and the highest sequence identity (99.4%) was found with Vietnam/Binh21/2015 (Vietnamese strain).Genome-wide phylogenetic analysis showed that the PDCoV 1468B strain belonged to group Ⅱ and was closely related to the PDCoV strain in Southeast Asian countries which belonged to the same evolutionary branch.In summary,PDCoV 1468B strain was successfully isolated and sequenced,which would lay a foundation for further research on biological characteristics of PDCoV,such as pathogenicity and vaccine development,and provided data support for genetic evolution of PDCoV.

Key words: porcine deltacoronavirus (PDCoV); isolation and identification; complete genome; sequence analysis

CLC Number:

S852.65

QIN Yibin, HE Pingping, LU Bingxia, HE Ying, LIANG Jiaxing, LIU Lei, DUAN Qunpeng, WEI Pinyuan, LI Bin, CHEN Zhongwei, ZHOU Yingning, SU Qianlian, JIANG Dongfu, LU Jingzhuan, ZHAO Wu. Isolation, Identification and Complete Genome Sequence Analysis of Porcine Deltacoronavirus Strain CH/GX/1468B/2017[J]. , 2019, 46(7): 1907-1916.

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Isolation, Identification and Complete Genome Sequence Analysis of Porcine Deltacoronavirus Strain CH/GX/1468B/2017

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