摘要
目的 建立小鼠腺病毒（MAdV） PCR检测方法,应用于长爪沙鼠、小鼠等实验动物MAdV的检测.方法 根据已发表的小鼠腺病毒（MAdV） E1B基因序列,设计合成引物.建立RT-PCR方法,并对方法的特异性、敏感性、稳定性等进行验证.用建立的方法对65只长爪沙鼠、12只小鼠进行检测.结果 建立的MAdV PCR检测方法与小鼠微小病毒、多瘤病毒无交叉反应;以MAdVDNA为模板,所能检测DNA最小模板浓度为1.67 pg/μl,可检测病毒最小滴度为10-7/ml; MAdVDNA在-30℃冰箱放置12个月,仍能扩增出大小约606 bp的可见目条带.经PCR检测,65只沙鼠和12只小鼠均为阴性.结论 建立的小鼠腺病毒（MAdV） PCR检测方法特异、敏感、稳定,可用于小鼠、长爪沙鼠等实验动物MAdV的检测.
Objective To develop a RT-PCR method for detection of Mouse Adenovirus （MAdV） in Mongolian gerbils and Laboratory mice. Methods The primers were designed and synthesized according to the published MAdv specific sequences of E1B gene. PCR method is established, and the specificity, sensitivity, stability test were carded out. The method is used to detect 65 Mongolian gerbils, and 12 mice. Results The developed PCR method was no cross reaction with Minute virus, Polyoma virus, its minimum detection limit ,using the recombinant plasmid containing MAdV gene as a template, was 1.67 pg/ml, and the lowest detection virus titer is 10-7/ml. The MAdV cDNA maintained at -30℃ for 12 months, still can enlarge the size of about 606 bp visible mesh belt. The 65 Mongolian gerbils and 12 mice were all negative about MAdv detected by PCR. Conclusion The developed PCR method is good in specificity, sensitivity, stability,and may be used for detecting the MAdv in laboratory animal, such as Mongolian gerbils and mice.
出处
《实验动物与比较医学》
CAS
2014年第1期35-41,共7页
Laboratory Animal and Comparative Medicine
基金
国家科技支撑计划项目（2011BAI15801）
