云南边境地区帕利亚姆血清群中山病病毒的分离与鉴定

doi:10.16431/j.cnki.1671-7236.2022.11.031

摘要/Abstract

摘要： 【目的】了解西南边境地区牛感染中山病病毒（Chuzan virus，CHUV）的情况。【方法】应用BHK21细胞对云南景洪、江城采集的312份健康黄牛血液样品盲传分离病毒，对出现细胞病变的毒株进行形态学、基因组带型和分子生物学鉴定，对分离到的病毒进行S7、S2基因序列测定和比对分析。【结果】 4份血液样品可致BHK21细胞病变，电镜观察病毒颗粒呈球形，直径约50 nm；琼脂糖凝胶电泳发现，4株分离毒株基因组为10节段，带型为"3-3-4"，与2012年云南师宗分离的CHUV SZ187毒株相似；4株病毒S7、S2基因核苷酸序列相似性分别为98.7%~99.7%和97.9%~99.1%；S2基因片段核苷酸和氨基酸序列与中国本土分离的SZ187、GHN-GS-16等毒株相似性最高；S7基因片段核苷酸和氨基酸序列与日本分离的ON-1/E/18、ON-3/E/17及中国本土的SZ187、GHN-GS-16等毒株的相似性最高。S7、S2基因遗传进化分析结果显示，4株新分离毒株亲缘关系最近；4株毒株的S7基因序列与中国本土和日本分离的已知帕利亚姆血清群病毒（Palyam serogroup virus，PALV）毒株位于同一分支，提示4株毒株为PALV，且形成一簇，有一定的地域性；4株毒株S2基因片段核苷酸序列与中国本土分离的CHUV位于同一分支，亲缘关系较近，进一步证实这4株毒株为CHUV。【结论】本研究首次报告了从云南边境地区牛群中分离到CHUV毒株，为CHUV在中国的流行病学和边境地区疫病风险防控提供了重要参考及有力依据。

关键词: 中山病病毒(CHUV); 分离鉴定; 序列分析

Abstract: 【Objective】 This study was aimed to understand the situation of cattle infected with Chuzan virus (CHUV) in the southwest frontier area.【Method】 BHK21 cells were used to isolate virus from the blood samples of healthy cattle collected in Jinghong city and Jiangcheng county, Yunnan province.The samples with cytopathy were identified by morphology, genomic banding and molecular biology.The sequences of S7 and S2 genes were determined and compared.【Result】 Four samples with cytopathy were found.The virus particles were spherical with a diameter of about 50 nm by electron microscopy.The results of agarosegel electrophoresis showed that the genome of the four newly isolated viruses was 10 segments and the band was "3-3-4", which was similar to the strain of CHUV SZ187 isolated from Shizong in Yunnan province in 2012.The nucleotide similarity of the S7 and S2 genes of the four viruses were 98.7%-99.7%, 97.9%-99.1%.The sequence of S2 fragment had the highest homology with SZ187 and GHN-GS-16 strains isolated in China.The nucleotide and amino acid sequence of S7 gene had the highest homology with ON-1/E/18, ON-3/E/17 in Japan and SZ187, GHN-GS-16 in China.The results of genetic evolution analysis of S7 and S2 genes showed that the four newly isolated strains were the closest relatives.The S7 gene sequences of the four strains were in the same branch as the known PALV strains isolated from China and Japan, indicating that the four strains were PALV and formed a cluster, which had a certain regional character.The four strains were further confirmed to be CHUV, because the sequence of S2 gene of the four strains was on the same branch as the isolated CHUV from China.【Conclusion】 This study first reported the isolation of CHUV from cattle in the border area of Yunnan province.It provided an important reference and a powerful basis for the epidemiology of CHUV in China and the prevention and control of epidemic disease risk in border areas.

Key words: Chuzan virus(CHUV); isolation and identification; sequence analysis

中图分类号:

S852.65^＋9.4

寇美玲, 苗海生, 李乐, 谢佳芮, 李华春, 宋建领. 云南边境地区帕利亚姆血清群中山病病毒的分离与鉴定[J]. 中国畜牧兽医, 2022, 49(11): 4401-4409.

KOU Meiling, MIAO Haisheng, LI Le, XIE Jiarui, LI Huachun, SONG Jianling. Isolation and Identification of Chuzan Virus from Palyam Serogroup Virus in Yunnan Border Area[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4401-4409.

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