猪伪狂犬病病毒GZYY2015变异株的分离鉴定

doi:10.16431/j.cnki.1671-7236.2021.05.035

Abstract

Abstract: The purpose of this experiment was to isolate Pseudorabies virus (PRV) from a sample tested as pseudorabies (PR) sent by a pig farm in Yunyan district,Guiyang city,Guizhou province.Methods such as cytotoxicity experiments,physical and chemical experiments,and transmission electron microscopy observations were used to isolate and identify the pathogen,and then to determine the virus value of the pathogen and then conduct animal experiments and comparative analysis of gD and gE genes sequences.The results showed that the cell inoculation experiment successfully isolated PRV that could cause Vero cells to produce typical lesions.The physicochemical experiment showed that the isolated virus was unstable in the environment of acid and heat,and sensitive to idoxuridine(IUDR) and chloroform.Transmission electron microscopy results showed that the mature virus particles with envelope in nucleus were approximatively round with a diameter of 150-160 nm,and the virus particles in nucleus were hollow or stuffed without envelope,and the capsids were lattice arrangement in mature viral inclusion bodies.Animal experiment showed that the strain had strong pathogenicity to rabbits,which could cause itching at the inoculation site.The results of nucleic acid identification showed that the open reading frame(ORF) of the amplified PRV gD gene was 1 209 bp,and the ORF of gE gene was 1 740 bp.The results of GenBank comparison showed that the gD gene of the isolated strain was 99.8% similarity with the reference strains of PRV JS2012 strain (accession No.:KP257591) and PRV HNB strain (accession No.:KM189914.3),and the similarity with vaccine strain Bartha-K61 (accession No.:JF797217.1) was 98.7%.The gE gene nucleotide sequence was 99.9% similar to PRV JS 2012 strain (accession No.:KP257591),PRV HNB strain (accession No.:KM189914.3) and PRV Qihe547 (accession No.:KU056477).According to the phylogenetic tree of gD and gE amino acid sequences,the isolated strain belonged to the same branch of the domestic mutant strains.These results indicated that the PRV variant strain was successfully isolated,which could provide references on PRV epidemiology,genetic evolution analysis and immune control of PR in Guizhou province.

Key words: Pseudorabies virus; mutant strain; isolation and identification; gE gene; gD gene

CLC Number:

S852.65⁺1

HUANG Ersu, XIAN Wen, ZHANG Aiqiong, WANG Bin, ZENG Zhiyong, TANG Deyuan, LIANG Haiying, HE Xiaoli, XU Yu, XU Songping, ZHU Yang. Isolation and Identification of Porcine Pseudorabies Virus GZYY2015 Mutant Strain[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(5): 1832-1840.

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Isolation and Identification of Porcine Pseudorabies Virus GZYY2015 Mutant Strain

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