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〔目的〕 快速检测水中肠道病毒。〔方法〕 一步单管反转录 PCR,通用引物多重 PCR。〔结果〕 反转录与 PCR在一个反应管中一步操作即可完成 ,用 6条引物一次反应成功地检测了甲型肝炎病毒 (HAV)、脊髓灰质炎病毒 (Polio)、柯萨奇 B组病毒 (Cox B)和埃可病毒 (EV)。其特异性为半套式 PCR所证实 ,敏感性达 10 1 PFU / ml。〔结论〕 一步单管反转录 PCR,简化了操作步骤 ,减少了污染的机会 ,提高了反应的敏感性和特异性。通用引物多重PCR用较少的引物检测并鉴别较多的病毒 ,减少了引物之间的干扰 ,提高了结果的稳定性。
[Objective] Rapid detection of waterborne enterovirus. [Method] One-step single-tube reverse transcription PCR, universal primer multiplex PCR. [Results] Reverse transcription and PCR were performed in one reaction tube in a single step. Six reactions of HepA, Polio, Coxsackie B Cox B and Echovirus (EV). The specificity was confirmed by semi-nested PCR with a sensitivity of 101 PFU / ml. [Conclusion] One-step single-tube reverse transcription PCR simplified the procedure, reduced the chance of contamination and increased the sensitivity and specificity of the reaction. Universal primer multiplex PCR with fewer primers to detect and identify more viruses, reducing the interference between primers and improve the stability of the results.