猪轮状病毒江西株AY01的分离鉴定

doi:10.16431/j.cnki.1671-7236.2022.08.031

Abstract

Abstract: 【Objective】 The study was aimed to determine the etiology of diarrhea piglets in a pig farm in Jiangxi province.【Method】 The small intestine samples of piglets were tested for Porcine epidemic diarrhea virus (PEDV),porcine Transmissible gastroenteritis virus (TGEV) and Porcine rotavirus (PoRV) by RT-PCR.Positive sample was inoculated into MA104 cells for passage to isolate PoRV.The isolated strain was detected by electron microscope observation,indirect immunofluorescence test,PoRV VP4 and VP7 genes sequencing and animal regression test.【Result】 The results showed that porcine intestinal disease samples were incubated at 37 ℃ for 2 h with final concentration of 15 μg/mL trypsin and inoculated with MA104 cells,which could proliferate and passage on cells,and the 6th-generation began to show stable cytopathic effect. Electron microscopic observation showed that the virus particles were 61-70 nm in diameter,the average size was 65 nm,with short fibrils and smooth outer edges,similar to wheel-like particles,which had typical morphological characteristics of PoRV virus particles.Both indirect immunofluorescence assay and RT-PCR were positive for PoRV,and the isolated strain was determined to be PoRV.Sequence analysis of VP4 and VP7 genes revealed that VP4 genotype had the highest homology with P【23】 genotype, and VP7 genotype had the highest homology with G5 genotype,according to the latest classification method of group A Rotavirus,the isolate was classified as G5P【23】 genotype.The results of animal regression test showed that the one-day-old newborn piglets with oral infection of the isolate had watery diarrhea,vomiting and other clinical symptoms about 24 h after infection,and could detect PoRV in the feces.【Conclusion】 Through continuous passage of MA104 cells,a strain of PoRV was successfully isolated from the small intestine samples of diarrhea piglets in a pig farm in Jiangxi province,the isolated strain belonged to G5P【23】 genotype PoRV,which was the pathogen of diarrhea in piglets.

Key words: Porcine rotavirus (PoRV); isolation and identification; animal regression experiment

CLC Number:

S852.65⁺1

LIU Xiaolan, LIU Changjin, YU Wenyang, LI Xiaoxiang, BIAN Yanchao, HUANG Xiaohua, LUO Feng, DENG Shunzhou. Isolation and Identification of Porcine Rotavirus Jiangxi Strain AY01[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 3151-3162.

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Isolation and Identification of Porcine Rotavirus Jiangxi Strain AY01

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