竹鼠源细小病毒分离鉴定及全基因组序列分析

doi:10.16431/j.cnki.1671-7236.2023.07.024

Abstract

Abstract: 【Objective】 This study was aimed to understand and master the biological characteristics and genetic variation of Bamboo rat parvovirus (BRPV) in Guangxi, and to provide theoretical basis and technical support for the control of BRPV.【Method】 The tissue supernatant was prepared from the diseased bamboo mice from farms, and the virus was isolated by Vero cell synchronous inoculation method.Identification was carried out through methods such as cytopathic observation, PCR, transmission electron microscopy observation, and hemagglutination test, and specific amplification primers were designed and synthesized for sequencing analysis of the whole genome sequence of BRPV.【Result】 The isolated virus could grow and proliferate stably on Vero cells, and the infected cells became long, spindle or thread-like.After purification, virions with a diameter of 20-25 nm were observed by electron microscopy with stereosymmetric, membraneless virions.They were identified as BRPV by PCR, agglutination test and whole genome sequencing.In this study, a total of 3 BRPV (GenBank accession No.:MF497824, MF497825 and MF497826), the total length of the genome of this strain was about 4 758 bp, and the whole genome sequence comparison showed that it was closely related to Bat-derived arvovirus.The amino acid genetic characteristics of BRPV gene have strong correlation with its host specificity.The amino acid variation of NS1 gene was more similar to that of bat and Rat parvovirus, which was in the same branch.Nucleotide homology was 96.9% to 97.6%, and amino acid homology was 97.5% to 98.4%.Amino acids 257 and 360 belonged to high mutation sites.The phylogenetic tree of VP2 gene encoded amino acids showed that the phylogenetic tree was located in an independent branch, with the nucleotide homology of 58.3% to 66.8% and the amino acid homology of 51.6% to 63.0% with rat, dog, cat and other animal parvoviruses.Amino acids at sites 8, 99, 114, 118, 247 and 253 of VP2 peotein showed host bias, and amino acids at sites 53 and 386 were highly mutated.【Conclusion】 The BRPV strains isolated in this study were Parvovirus.The results clarified the genetic characteristics and morphological characteristics of BRPV in China, which laid a foundation for the epidemiological investigation and control of BRPV.

Key words: Bamboo rat parvovirus (BRPV); isolation and identification; whole genome; genetic evolutionary analysis

CLC Number:

S852.65

TANG Haibo, JIANG Jiajia, CHEN Fenglian, YANG Jinlan, BAI Anbin, LIU Jinfeng, QIN Shaomin, WU Jianmin. Isolation, Identification and Genomic Characterization Analysis of Parvovirus Isolate from Bamboo Rat[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(7): 2843-2853.

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Isolation, Identification and Genomic Characterization Analysis of Parvovirus Isolate from Bamboo Rat

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