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志贺菌依赖解旋酶DNA恒温扩增技术建立

Detection of Shigella with helicase-dependent isothermal DNA amplification technology

  • 摘要: 目的 利用依赖解旋酶DNA恒温扩增技术(HDA),建立一种快速检测志贺菌(Shigella)的新方法。方法 根据志贺菌的ipaH基因序列设计特异性引物,优化反应体系和反应条件;并对方法进行特异性和灵敏度评价。结果 对21株实验菌株检测,3株志贺菌均为阳性,其余18株非志贺菌均为阴性,灵敏度为5.1×103cfu/mL,与普通PCR方法检测结果相当。结论 HDA法检测志贺菌具有特异、灵敏及仪器要求低等特点,具有广阔的应用前景。

     

    Abstract: Objective To establishe a new rapid method to detect Shigella based on helicase-dependent isothermal DNA amplification(HDA).Methods A highly specific set of primers was synthesized to target ipaH gene of Shigella and then HDA condition and the reaction system were optimized simultaneously.Specificity and sensitivity of the method w ere evaluated.Results The results of all three strains of Shigella were positive,and the othes were negative.The sensitivity was 5.1 × 103 cfu/mL,which was similar to the result of PCR method.Conclusion Detecting Shigella with HDA is specific and sensitive as PCR method and has lower instrumental requirement.

     

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