摘要
目的:建立两种简单、快速和特异性的RT-PCR法鉴定SARS-HCoV、HCoV-229E和HCoV-OC43。方法:用DNA Star和Prem ier 5.0软件设计SARS-HCoV、HCoV-229E和HCoV-OC43及其它冠状病毒针对保守区Pol1b基因的一对通用引物和分别针对M基因的3对引物,然后通过对扩增片断测序、限制性酶切方法和根据扩增片断的长度进行鉴定。结果:两种方法均能扩增到与预期目标一致的片断,特异性和灵敏度高,能快速区分3种病毒。结论:这些方法将为了解冠状病毒在呼吸道感染中的作用提供一个有利的工具。
Objective：To establish two simple, rapid and specific methods to diagnose human coronaviruses：SARS- HCoV, HCoV - 229E and HCoV - OC43. Methods： The primers of the assay were designed with the software packages of DNA star and primer premier 5.0. The first method was based on consensus primers targeting conserved regions Pollb gene and the second method on consensus primers targeting conserved regions M gene of coronavirus genome sequences. Species identification was provided by sequencing the amplicons or by a rapid screening test by restriction enzyme analysis or by the length of the amplicons. Results： Two methods could be used to detect three coronavirus with high specificity and sensitiveness. Conclusion：These methods could provide a useful tool for demonstrating the role of human coronavirus in infection of the respiratory tract.
出处
《中国卫生检验杂志》
CAS
2006年第5期513-515,553,共4页
Chinese Journal of Health Laboratory Technology
基金
广东省科技攻关重大项目(SARS项目)资助(2003FD02-05)
