【摘要】 目的 研制一项检测严重急性呼吸综合征的即时逆转录-聚合酶链反应(RT-PCR)方法,用于患者全血的检查。方法 内引物和短柄圆环探针(分子信标,molecular beacon)的设计,针对基因库SARS相关冠状病毒的高度保守区(RNA聚合酶基因区),在15 301和15 480之间(G130027616)。结果 即时RT-PCR荧光测量结果能显示SARS相关冠状病毒特有序列的扩增程度。常规PCR的145bp产物经测序证实与探针杂交的靶序列一致。结论 这一方法的建立对SARS临床诊断和病原研究提供一种快捷和准确的手段。更多还原

【Abstract】 Objective To develop a newly real-time RT-polymerase chain reaction assay for severe acute respiratory syndrome (SARS) related coronavirus in human whole blood. Methods A pair of primers and a probe ( molecular beacon) had been designed that were specific for the recognition of a highly conservative region between 15 301 and 15 480 of the SARS-related coronavirus polymerase gene sequences obtained from GenBank ( G130027616). Results In the real-time RT-PCR assay, the extent of SARS related coronavirus amplification was measured in terms of the increase in fluorescence during the amplification process. The 145 bp fragment of PCR product was further confirmed by conventional PCR assay and proved by DNA sequencing to be identical to the target sequence to which the probe was hybridized. Conclusion This assay has a broad application for clinical diagnosis and surveillance investigation.更多还原