文章摘要
重组酶介导等温扩增技术检测哈维氏弧菌
Rapid detection of Vibrio harvey with Recombinase Aided Amplification
投稿时间:2020-05-23  修订日期:2020-11-16
DOI:
中文关键词: 哈维氏弧菌  重组酶介导等温扩增  快速检测
英文关键词: Vibrio harvey  Recombinase aided amplification  Rapid detection
基金项目:国家科技重大专项“海上丝绸之路及国门输入重要传染病军民融合防控技术研究”(2018ZX10101003);病原微生物生物安全国家重点实验室开放研究基金资助项目(SKLPBS1832)
作者单位邮编
方微微 上海海洋大学 201306
卢瑛 上海海洋大学 201306
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中文摘要:
      摘要:针对哈维弧菌LuxS基因保守区段设计特异性引物、探针,通过引物、探针筛选以及反应条件优化,建立基于重组酶介导等温扩增(recombinase-aided amplification, RAA)快速、灵敏、特异检测哈维弧菌的方法。,采用细菌纯培养物评估该方法的最低检测限、特异性等性能,通过对加标样品检测,与LAMP,PCR等方法比较评估该检测方法的可靠性。结果显示该方法在 39 °C条件下 20 min内可完成检测,检测细菌纯培养物的最低检测浓度为7.55×10-1 copies/μL,且与其他食源性致病菌无交叉反应,特异性良好。综上,本文建立的哈维弧菌重组酶介导等温扩增检测方法快速、特异、简便、灵敏,在食源性疫情现场调查以及食源性疾病监测中有一定的应用前景。
英文摘要:
      The purpose of this study was to develop a rapid, simple and sensitive diagnostic test based on RAA to detecting V. harvey. Specific primers and probes were designed for the conserved section of the LuxS gene of V.harvey. Through primer and probe screening and optimization of reaction conditions, a fluorescent RAA method for detecting V. harveyi was established. Use the pure bacterial culture to evaluate the minimum detection limit, specificity and other performance of the method, and test it by simulating the sample, and use the LAMP, PCR and other methods to evaluate the reliability of the method. Our results indicated that the method can complete the detection within 20 min at 39 °C. No positive reaction was observed besides of pathogenic V. harvey. 7.55×10-1 copies/μL V.harvey could be detected within 20 min. This recombinase-aided amplification method for the detection of V. harvey is fast, specific, simple and sensitive, and shows a potential application in the investigation of foodborne outbreaks and surveillance of foodborne diseases of V. harvey.
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