志贺氏菌基于核酸序列恒温扩增检测方法研究

Study in the Nucleic Acid Sequence-Based Amplification Detecting Method of Shigella

使用自行建立和优化的基于核酸序列恒温扩增(Nucleic Acid Sequence-based Amplification,NASBA)检测体系,对志贺氏菌(Shigella spp.)进行检测。采用志贺氏菌的ipaH基因为目的片段设计特异性引物,建成可快速检测志贺氏菌的NASBA检测法,并进行特异性和灵敏度试验。结果表明:该检测方法具有较高的灵敏度和可靠的特异性,灵敏度为6.3×102cfu/mL,高于普通PCR方法。

A new method,which is based on Nucleic Acid Sequence Amplification（NASBA） to detect Shigella of sample,was established.A specific set of primers was synthesized to target the ipaH gene of Shigella so as to establish Nucleic Acid Sequence-based Amplification method.Specific and sensitivity were tested.The results showed that the sensitivity of NASBA was 6.3×102 cfu/ml which was higher than the result of PCR method.NASBA was more specific and sensitive than PCR method and has lower instrumental requirement.