牛A群轮状病毒G10型XJ-2022株分离鉴定及基因型分析

doi:10.16431/j.cnki.1671-7236.2022.09.027

Abstract

Abstract: 【Objective】 The purpose of the experiment was to identify the etiology and genotype of calves suffering from diarrhea in a large-scale cattle farm in Xinjiang.【Method】 15 fecal samples were randomly collected from calves suffering from diarrheal diseases in a cattle farm in Xinjiang using the antigen diagnostic kit method.The virus was isolated and passaged in Marc-145 cells.Indirect immunofluorescence assay (IFA) and negative-staining direct electron microscopy were performed on the isolated strains to further identify the pathogen.The VP6 and VP7 genes of the isolates were amplified and sequenced by PCR and genetic evolution analysis.【Result】 The results of the antigen diagnostic kit showed that 3 stool samples were positive for Bovine rotavirus (BRV) antigen.Positive fecal samples were inoculated into Marc-145 cells, and only 1 sample showed obvious cytopathic effect (CPE) in the ninth passage.The IFA results showed that the Marc-145 cells inoculated with this isolate had bright green fluorescence, but no fluorescence was seen in the control group.Circular virus particles of about 65 nm were observed by electron microscope, and were named as XJ-2022 strain.The target bands of VP6 and VP7 genes were obtained by PCR amplification with lengths of 1 356 and 342 bp, respectively.Gene similarity comparison and genetic evolution analysis showed that VP6 gene had the highest similarity and the closest genetic evolutionary relationship with the human group A Rotavirus reference strain DB2015-066 (LC367318.1), and VP7 gene had the highest similarity and genetic evolutionary relationship with bovine G10 Rotavirus reference strain XJX2(MN937506.1), the isolate was identified as group A G10 Rotavirus.【Conclusion】 The genotype of group A G10 BRV XJ-2022 strain was successfully isolated, which was the first multi-host gene reassortment virus discovered in Xinjiang.

Key words: Bovine rotavirus (BRV); isolation and identification; IFA; genotyping

CLC Number:

S852.65^＋3

XI Jing, YI Jihai, WANG Yueli, XU Zhenyu, LI Peidong, ZHANG Jiangwei, CHEN Chuangfu. Isolation, Identification and Genotype Analysis of Group A Bovine Rotavirus G10 Strain XJ-2022[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3530-3538.

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Isolation, Identification and Genotype Analysis of Group A Bovine Rotavirus G10 Strain XJ-2022

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