论文部分内容阅读
目的 探索SARS冠状病毒早期诊断的有效手段。方法 对临床病例样品采用反转录巢式PCR方法鉴定SARS冠状病毒。提取样品RNA,用巢式PCR外侧下游引物进行反转录,以反转录产物为模板,进行巢式第1次、第2次PCR。PCR产物克隆到pMD18-T载体后进行测序鉴定。结果 被检样品扩增出特异片段,经测序验证确实来自SARS相关冠状病毒。Blast比较结果与SARS冠状病毒多伦多株相应片段只有一个碱基的差异。结论 反转录巢式PCR方法验证在非典型肺炎患者标本中存在SARS相关冠状病毒的序列,证实该法不失为一种检测SARS冠状病毒的快速、简便、易行的方法。
Objective To explore the effective means of early diagnosis of SARS coronavirus. Methods The SARS coronavirus was identified by reverse transcription nested PCR in clinical cases. The RNA samples were extracted and reversely transcribed with primers downstream of the nested PCR. The first and second nested PCR were performed using the reverse transcripts as a template. The PCR product was cloned into pMD18-T vector and sequenced. Results The samples were amplified specific fragments, confirmed by sequencing confirmed from SARS-associated coronavirus. The result of Blast is only one base difference from the corresponding fragment of the SARS coronavirus strain Toronto. Conclusion The reverse transcriptional nested PCR method was used to verify the sequence of SARS-associated coronavirus in SARS patients, which confirmed that this method is a rapid, simple and easy method to detect SARS coronavirus.