摘要
目的:建立食品中脊髓灰质炎病毒的RT-PCR检测体系。方法:在脊髓灰质炎病毒基因组2A区保守序列设计引物,建立同时适于3种血清型脊髓灰质炎病毒的检测方法,并对人工接种病毒的贝类(生蚝、文蛤)、蔬果(草莓、樱桃和生菜)和水样等实际样品进行检测。结果:基于构建的分别适于3种血清型脊髓灰质炎病毒检测的参照分子,确定所建立方法的检测下限可达50拷贝参照分子DNA。针对各种食品基质建立的脊髓灰质炎病毒富集、RNA提取和RT-PCR方法检测3种血清型病毒可达10RT-PCR单位(RTU)。结论:建立的RT-PCR检测体系可用于食品中脊髓灰质炎病毒的检测。
Objective：To establish a RT-PCR system suitable for polioviruses detection in foodstuffs.Methods： A primer pair based on the conserved 2A region of poliovirus genomic cDNA has been designed to set up the detection system for three serotypes of polioviruses.Moreover,several inoculated samples,including oyster,clam, strawberry,cherry,lettuce and water were analyzed by the developed detection system.Results：Sensitivity tests based on three constructed reference molecules showed that 50 copies of reference molecules DNA can be stably detected.Results from inoculated samples analyses suggested that the concentration of 10 RTU of either serotype 1, 2 or 3 of poliovirus can be detected in six food samples using the optimized virus concentration,RNA extraction and RT-PCR methods.Conclusion：The established RT-PCR system can be applied to the detection of polioviruses in foodstuffs.
出处
《生物技术通讯》
CAS
2010年第1期65-69,共5页
Letters in Biotechnology
基金
上海技术标准专项(07DZ05026)
国家质检总局科研项目(2007B150)
科技部世博科技专项(2009BAK43B31)
