Vabret A, Mouthon F, Mourez T, Gouarin S, Petitjean J, Freymuth F
Laboratory of Human and Molecular Virology, University Hopital, av. Georges Clemenceau, 14 033, Caen, France.
J Virol Methods. 2001 Sep;97(1-2):59-66. doi: 10.1016/s0166-0934(01)00343-3.
An RT-PCR-hybridization was developed that amplified genetic material from the M protein gene of HCoV-229E and HCoV-OC43. The analytic sensitivity of these original primers were compared with primers defined in the N gene and described previously. The results show that 0.05 TCID50 of HCoV-229E and 0.01 TCID50 of HCoV-OC43 can be detected by this molecular method using the original method. Detection of HCoV-229E and HCoV-OC43 in clinical specimens is possible using this method: 348 respiratory specimens (202 sputum and 146 nasal aspirates) were tested with this RT-PCR-hybridization and 12 human coronavirus are detected (3%). The method could provide a useful tool for demonstrating the role of human coronavirus in infections of the respiratory tract.
开发了一种逆转录聚合酶链反应-杂交方法,该方法可扩增人冠状病毒229E(HCoV-229E)和人冠状病毒OC43(HCoV-OC43)M蛋白基因的遗传物质。将这些原始引物的分析灵敏度与先前描述的N基因中定义的引物进行了比较。结果表明,使用原始方法,通过这种分子方法可检测到0.05半数组织培养感染剂量(TCID50)的HCoV-229E和0.01 TCID50的HCoV-OC43。使用该方法可检测临床标本中的HCoV-229E和HCoV-OC43:用这种逆转录聚合酶链反应-杂交方法检测了348份呼吸道标本(202份痰液和146份鼻吸出物),检测到12株人冠状病毒(3%)。该方法可为证明人冠状病毒在呼吸道感染中的作用提供有用的工具。