CAJ | 학술논문

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RT-PCR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。
목적：료해상해지구실험소서자연감염소서낙와극병독（ murine norovirus ，MNV）적상황，병분리독주。방법추취위탁검측단위송검적SPF소서319지，분별채집맹장내용물급혈액양본，응용역전록-취합매련반응（ RT-PCR）방법확증소서맹장내용물양본중MNV적특이성기인편단래검측MNV적감염정황，동시채용매련면역흡부시험（ enzyme linked immunosorbent assay ，ELISA）여핵산검측방법진행대비。장RT-PCR확증결과위양성적맹장내용물양본희석병경0.22μm려막과려，접충도소서거서세포계RAW 264.7세포，맹전후채용RT-PCR방법감정。결과 RT-PCR검측적319빈소서맹장내용물양본중，양성양본95빈，양성솔위29.78％。대180빈경RT-PCR검측적소서혈청진행ELISA검측，양성양본70빈，양성솔위38.89％。 RAW 264.7세포맹전5대후재72 h내출현세포병변，경RT-PCR감정，현시187 bp적목적조대。결론통과핵산검측방법화혈청학방법증실상해지구실험소서존재MNV자연감염，차감염솔교고，응가강실험소서적사양관리。
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 .