上海地区小鼠诺瓦克病毒的检测分析及病毒分离  

Detection and isolation of murine norovirus in laboratory mice in Shanghai area

作　　者：刘芹[1,2,3] 魏晓锋 田立立 高诚[2,3] 

机构地区：[1]复旦大学,上海200032 [2]上海市计划生育科学研究所,上海200032 [3]上海实验动物研究中心,上海201203

出　　处：《中国实验动物学报》2014年第2期80-85,I0009,共7页Acta Laboratorium Animalis Scientia Sinica

基　　金：上海市科委创新行动计划"小鼠诺瓦克病毒的特性研究"(NO.12140900502)

摘　　要：目的了解上海地区实验小鼠自然感染小鼠诺瓦克病毒(murine norovirus,MNV)的状况,并分离毒株。方法抽取委托检测单位送检的SPF小鼠319只,分别采集盲肠内容物及血液样本,应用逆转录-聚合酶链反应(RT-PCR)方法扩增小鼠盲肠内容物样本中MNV的特异性基因片段来检测MNV的感染情况,同时采用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)与核酸检测方法进行对比。将RT-PCR扩增结果为阳性的盲肠内容物样本稀释并经0.22μm滤膜过滤,接种到小鼠巨噬细胞系RAW 264.7细胞,盲传后采用RT-PCR方法鉴定。结果 RT-PCR检测的319份小鼠盲肠内容物样本中,阳性样本95份,阳性率为29.78%。对180份经RTPCR检测的小鼠血清进行ELISA检测,阳性样本70份,阳性率为38.89%。RAW 264.7细胞盲传5代后在72 h内出现细胞病变,经RT-PCR鉴定,显示187 bp的目的条带。结论通过核酸检测方法和血清学方法证实上海地区实验小鼠存在MNV自然感染,且感染率较高,应加强实验小鼠的饲养管理。Objective To investigate the natural infection status of murine norovirus （ MNV） in laboratory mice in Shanghai area and isolate MNV from mouse cecal feces .Methods To collect cecal contents and serum samples from 319 specific pathogen-free （ SPF） mice coming from different research institutions .Reverse transcription-polymerase chain reaction （ RT-PCR） and enzyme linked immunosorbent assay （ ELISA） were used to detect MNV infection in the mice , re-spectively.The positive stool samples were diluted and filtered through 0.22 μm membrane, inoculated into RAW 264.7 cells, and then identified by RT-PCR.Results There were 95 positive results in the 319 cecal samples by RT-PCR, and the positive rate was 29.78%.Among 180 serum samples which were tested by RT-PCR, 70 samples were positive by ELISA, and the positive rate was 38.89%.The infected RAW 264.7 cells showed cytopathic effect （ CPE） within 72 h. After 3 times of freezing and thawing , RT-PCR obtained a 187 bp band.Conclusions The results from the present study show that there is a high natural infection rate of MNV in laboratory mice in Shanghai area , and the strict breeding manage-ment must be strengthened .

关 键 词：小鼠诺瓦克病毒 自然感染 逆转录-聚合酶链反应 酶联免疫吸附试验 

分 类 号：Q95-33[生物学—动物学] R285.5[医药卫生—中药学]