应用一步和套式PCR法检测SARS患者标本中的SARS病毒核酸序列

目的：建立敏感、特异和快速的针对SARS病毒的RT-PCR方法，为SARS的早期诊断及防治提供依据。方法：采用一步和套式PCR法对SARS患者不同标本中的SARS病毒RNA进行扩增及序列测定。结果与结论：应用外引物和内引物可分别扩增出约380bp和150bp的单一条带，其大小与预期的相一致。应用套式PCR可使检测敏感性从l0-6提高到lO-15。采用这种方法从SARS患者粪便、痰、鼻咽拭子和血液标本中均可扩增出与预期大小一致的条带，且测序结果显示该扩增片段为SARS病毒的特异序列，表明本研究建立的RT-PCR方法敏感、特异，可用于SARS的快速检测.．

Detection of SARS viruses in specimens from SARS patients by one-step and nested PCR assays

Objective:To develop a sensitive, specific and rapid RT_PCR assay for early diagnosis, effective prevention and treatment of SARS.Methods:cDNA fragments of SARS viruses from different specimens of SARS patients were amplified by one_step RT_PCR and nested PCR assays, then sequenced.Results and Conclusions:A single fragment with about 380?bp was amplified by outer primers and 150?bp fragment amplified by inner primers.The sizes of the amplified fragments were equal to those of the expected products. Nested PCR made the sensitivity of detection increase from 10 _6 to 10 _15 . The fragments with expected size could be amplified from stools, sputa, nasopharyneal swabs and sera of SARS patients, and the results of sequence determination showed that these amplified fragments were specific to SARS viruses, which indicated that the RT_PCR assay established in this study was sensitive and specific, and could be used for rapid detection of SARS.