【作者】 孟钰榕； 郑龙； 祝岩波； 王璇； 尤红煜； 刘健敏； 栗彦宁； 连伟光； 张东明； 王俊霞；

【Author】 MENG Yu-rong;ZHENG Long;ZHU Yan-bo;WANG Xuan;YOU Hong-yu;LIU Jian-min;LI Yan-ning;LIAN Wei-guang;ZHANG Dong-ming;WANG Jun-xia;Department of key laboratory Animal Hebei Province, laboratory Animal Hebei Medical University;The second hospital of Hebei Medical University;

【机构】 河北省实验动物重点实验室河北医科大学实验动物学部； 河北医科大学第二医院；

【摘要】 目的建立快速检测实验大鼠冠状病毒和仙台病毒的双重PCR方法。方法根据大鼠冠状病毒N基因、仙台病毒L基因设计特异性引物;经过双重PCR优化,特异性和敏感性的检测,建立双重PCR体系。应用该PCR体系检测人工感染样本和实验动物组织样本,并与ELISA方法比对。结果双重PCR扩增出大鼠冠状病毒(168bp)和仙台病毒(262bp)目的条带。仙台病毒和大鼠冠状病毒的检测下限为1.56×10~2 copies/μL。应用建立的双重PCR体系检测人工感染样本,30份仙台病毒核酸阳性标本均被检出。结论建立的双重PCR方法操作简单、快速、特异性强、灵敏度高,能够实现对实验动物仙台病毒和大鼠冠状病毒病原体的快速检测。更多还原

【Abstract】 Objective The purpose of this study is to establish a rapid duplex polymerase chain reaction(PCR) test for coinstantaneous identification ofRat Coronavirus and Sendai Virus.Methods Designing specific primers based on rats coronavirus N gene, sendai virus L gene. To establish duplex PCR system by optimizing concentration of primers and annealing temperature,testing specificity and sensitivity. Screening sample of artificial infections and experimental animal tissues through the PCR system that is compared with ELISA method. Results Duplex PCR amplification of Rat Coronavirus(168 bp) and Sendai Virus(262 bp).The lower limit of detection for Rat Coronavirus and Sendai Virusis 1.56×10~2 copies/μL.The specificity of detection was not detected from other pathogens.To test artificial infection specimens applying this newduplex PCR,30 Sendai Virus positivespecimenswere detected.Conclusions This essay established the duplex PCR method which has the superiority of specific,sensitive,simple and rapid,and provides a credible way for quick test in laboratory animals microbiology.更多还原