成都市猫诺如病毒的检测及VP1基因序列分析  

Detection of Feline Norovirus in Chengdu and Its VP1 Gene Sequence Analysis

作　　者：黄思琦 黄坚 杨晓农[1] 郭琪 李妍 HUANG Si-qi;HUANG Jian;YANG Xiao-nong;GUO Qi;LI Yan(College of Animal Science and Veterinary Medicine,Southwest Minzu University,Chengdu,Sichuan,610041,China)

机构地区：[1]西南民族大学畜牧兽医学院,四川成都610041

出　　处：《动物医学进展》2021年第1期25-32,共8页Progress In Veterinary Medicine

基　　金：西南民族大学研究生创新型科研项目(CX2019SZ104);西南民族大学中央高校基本科研业务费专项项目(2020NQN32);中央高校基本科研业务费专项项目(2019NQN41)。

摘　　要：为了解四川成都市猫诺如病毒(FNoV)的感染情况,应用公开报道的3对PCR引物对127份临床采集的猫粪便样本(含临床腹泻样本71例,临床健康样本56例)进行FNoV核酸检测,并根据病毒基因片段序列构建遗传进化树。结果显示,采用猫诺如病毒特异性引物FNoV-F9/R15的检出率为9.45%(12/127),高于采用杯状病毒广泛性引物p289d-p290d(2.36%,3/127)及诺如病毒特异性引物JV12Y/13I的检出率(0.79%,1/127)。检测发现在临床阳性样本中存在猫泛白细胞减少症病毒(FPV)、猫肠道冠状病毒(FECoV)和猫星状病毒(FAstV)的混合感染。12株临床阳性样本病毒的RNA聚合酶基因序列分析显示,它们与GenBank上提交的猫诺如病毒及狮子诺如病毒处于同一分支,证实为猫诺如病毒。基于VP1基因序列分析结果显示,临床毒株间核苷酸同源性为89.0%~100.0%,氨基酸同源性为96.5%~100%,均位于诺如病毒GⅣ.2分支。临床毒株可分为2个分支,其中4株与日本毒株(GenBank登录号:LC389583)、美国毒株(GenBank登录号:JF781268)关系紧密;而另外3株则独立聚为1个分支,并存在共有的氨基酸变异位点。结果表明,成都市猫群中存在FNoV的流行,并与其他猫肠道病毒呈现混合感染。采用引物FNoV-F9/R15检测灵敏度高,适用于FNoV的临床检测。由于FNoV基因不断变异,存在临床发病风险,需要对其进行持续监测。In order to understand the infection of feline norovirus(FNoV)in Chengdu city,127 clinical feline fecal samples(including 71 diarrhea samples and 56 clinical healthy samples)were detected with 3 pairs of PCR Primers,and the genetic evolution tree was constructed based on the sequence of viral gene 1(VP1)fragments.The results showed that the detection rate of norovirus specific primer FNoV-F9/R15 was 9.45%(12/127),higher than that of calicivirus universal primer p289d-p290d(2.36%,3/127)and norovirus specific primer JV12Y/13I(0.79%,1/127).Co-infection of feline leukopenia virus(FPV),feline enteric coronavirus(Fecov)and feline astrovirus(FAstV)was detected in FNoV positive samples.The RNA polymerase gene sequence analysis showed that 12 strains were in the same branch belonging to the feline norovirus and lion norovirus.Based on the analysis of VP1 gene sequence,the nucleotide homology and amino acid homology among 12 strains were 89.0%-100.0%and 96.5%-100%,all of which were located in the G-IV.2 branch of Norovirus.These strains can be divided into two branches,four of which are closely related to the Japanese strain(GenBank accession number:LC389583)and the American strain(GenBank accession number:JF781268),while the other three are isolated and have common amino acid variation sites.The results showed that FNoV was prevalent in feline population in Chengdu,and co-infected with other feline enteroviruses.The primer FNoV-F9/R15 is highest sensitive and suitable for the clinical detection for FNoV.The results attributed to continuous variation of the FNoV gene,constant monitoring is warranted.

关 键 词：猫诺如病毒 病毒性腹泻 反转录-聚合酶链反应 VP1基因 

分 类 号：S852.659.1[农业科学—基础兽医学] S858.293[农业科学—兽医学]