摘要
为了解广东省规模化猪场中猪轮状病毒(PoRV)流行毒株的基因特征,本试验在广东省某猪场采集腹泻仔猪肠道组织样品,经实时荧光定量PCR(qPCR)进行病毒初步鉴定;分离获得病毒后,通过间接免疫荧光、电镜观察进一步鉴定,并对分离毒株进行全基因组高通量测序和遗传进化分析。结果显示,qPCR初步鉴定该腹泻仔猪病料为PoRV感染,从阳性病料中分离获得1株能引起典型细胞病变的毒株,在电镜下可观察到似车轮状的病毒颗粒,在倒置荧光显微镜下观察有绿色荧光,将该PoRV分离毒株命名为GD2022,其完整基因型为G9-P[23]-I5-R1-C1-M1-A8-N1-T1-E1-H1;基因遗传进化分析结果显示,GD2022为人轮状病毒、猪轮状病毒和狗轮状病毒基因组重组后的毒株。本试验结果丰富了PoRV基因组学和分子流行病学相关的研究资料,为轮状病毒防控提供了数据支持。
To understand the genetic characteristics of prevalent strains of porcine rotavirus(PoRV) in a large-scale pig farms in Guangdong province,this study collected intestinal tissue samples from piglets with diarrhea in a pig farm,and the virus was preliminarily identified using quantitative real-time PCR(qPCR).After isolation,the virus was further identified by indirect immunofluorescence and electron microscopy,followed by whole genome next generation sequencing and genetic evolutionary analysis of the isolated strains.The results showed that the preliminary qPCR identified the diarrheal piglet samples as being infected with PoRV,and one strain capable of inducing typical cytopathic effects was isolated from positive samples,showing wheel-like virus particles under electron microscopy and green fluorescence under inverted fluorescence microscopy.The isolated PoRV strain was named GD2022,with a complete genotype of G9-P[23]-I5-R1-C1-M1-A8-N1-T1-E1-H1.Genetic evolutionary analysis revealed that GD2022 was a recombinant strain of human rotavirus,porcine rotavirus,and canine rotavirus genomes.These findings enrich the genomic and molecular epidemiological research data on PoRV,and provide data support for prevention and control of rotaviruses.
作者
樊高
申翰钦
王连想
马保华
周庆丰
李群辉
李薇
林丽苗
蔡新斌
张辉华
FAN Gao;SHEN Han-qin;WANG Lian-xiang;MA Bao-hua;ZHOU Qing-feng;LI Qun-hui;LI Wei;LIN Li-miao;CAI Xin-bin;ZHANG Hui-hua(College of Life and Engineering,Foshan Institute of Science and Technology,Foshan 528231,China;Research Institute of Guangdong Wen's Food Group Co.,Ltd.,Xinxing 527400,China;Foshan Customs Comprehensive Technology Center,Foshan 528010,China)
出处
《中国兽医杂志》
CAS
北大核心
2023年第6期47-54,共8页
Chinese Journal of Veterinary Medicine
基金
广州海关科研项目(2022GZCK04)
广东省科技专项资金(江科[2021]183号)。
关键词
猪轮状病毒(PoRV)
全基因组测序
腹泻
电镜观察
porcine rotavirus(PoRV)
whole genome sequencing
diarrhea
electron microscope observation
