鸭坦布苏病毒分离鉴定及其囊膜蛋白遗传进化分析

doi:10.16431/j.cnki.1671-7236.2022.04.032

Abstract

Abstract: 【Objective】 The aim of this study was to understand and grasp the epidemiological and biological characteristics of Duck Tembusu virus (DTMUV),in order to provide theoretical basis and technical support for DTMUV prevention and treatment.【Method】 The pathogen was isolated from 10 ducks showing typical clinical signs of DTMUV from duck farm in Hebei province by cell and chicken embryo inoculation method,and was identified by RT-PCR,transmission electron microscopy,Western blotting and indirect immunofluorescence assay (IFA).The animal regression test was performed to detect its virulency,and the inheritance of envelop protein was carried out.【Result】 The isolated DTMUV strain could well proliferate in DF-1 cells and produce typical cytopathic effect (CPE) and had ability to kill the chicken embryo.After purification,the virus particles with a diameter of 30-60 nm were observed under electron microscopy.RT-PCR results showed that there was a single band at about 270 bp, which was consistent with the expected size of DTMUV; Western blotting results showed that there was a specific band at 60 ku, which was consistent with the size of E protein; The results of IFA showed that after DF-1 cells were inoculated with virus, bright specific fluorescence appeared in the cytoplasm, the above results showed that the isolated virus was DTMUV.The identified virus strain was named as AX2020 strain.After intramuscular injection,the infection rate of AX2020 strain infected Peking duck was as high as 100%,the diseased ducks had typical clinical symptoms such as neurological symptoms and diarrhea.Through sequence alignment,it was found that AX2020 strain had the highest similarity with GA strain (MK907880.1),and was far away from SD14 strain (MH748542.1).Compared with commercial inactivated vaccine strain HB2010 (MN649262.1) and live vaccine strain FX2010 (MH414568.1),amino acid site mutations had occurred at positions 93,277 and 487 of AX2020 strain.【Conclusion】 A DTMUV strain AX2020 was successfully isolated,which showed strong pathogenicity in Peking ducks.Compared with the domestic vaccine strains,the envelope protein of AX2020 strain had mutation at amino acid site,which laid a certain foundation for epidemiological investigations and vaccine-related studies.

Key words: Duck Tembusu virus (DTMUV); isolation and identification; envelope protein; genetic evolutionary analysis

CLC Number:

S858.32

YIN Yajie, LIANG Ruiying, CUI Huan, MENG Lijia, NI Weiling, QIAO Sina, GUO Kangkang, ZHANG Bo, LI Songli, HOU Shaohua, DONG Shishan. Isolation,Identification of Duck Tembusu Virus,and Genetic and Evolutionary Analysis of Its Envelope Protein[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(4): 1497-1506.

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Isolation,Identification of Duck Tembusu Virus,and Genetic and Evolutionary Analysis of Its Envelope Protein

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