国境口岸志贺氏菌的NASBA快速检测  

Rapid detection of Shigella spp. using NASBA assay at frontier port

作　　者：钟响 左锋[2] 韩辉[3] 

机构地区：[1]烟台出入境检验检疫局,山东烟台264000 [2]天津出入境检验检疫局,天津300457 [3]中国检验检疫科学研究院,北京100176

出　　处：《口岸卫生控制》2014年第6期26-29,35,共5页Port Health Control

摘　　要：目的采用NASBA技术,建立国境口岸志贺氏菌的快速检测方法。方法根据志贺氏菌侵袭性质粒抗原ipa H基因设计引物,进行NASBA检测,并采用人工添加菌株污染样品进行验证。结果 NASBA技术检测志贺氏菌特异性强,能够从污染样品中扩增出391bp的特异性片段,试验中未出现假阳性和假阴性结果,灵敏度高,检出限达到8.2pg/μL总RNA;且检测周期短,NASBA反应在42℃90min即可进行有效扩增,24h即可完成样品中目标菌的筛选。结论因NASBA技术具有特异性强,灵敏度高,检测周期短等优点,不仅适用于国境口岸现场快速检测志贺氏菌,而且随着该技术的不断发展,必将有更为广阔的应用空间。Objective The rapid Shigella detecting method is established at frontier port using nucleic acid se- quence- based amplification （NASBA） assay. Methods A nucleic acid sequence -based amplification （NAS- BA） for the detection of Shigella is established using primers specific to ipaH gene, and artificially contaminated samples are used to verify the method. Results The assay can specifically amplify a 391bp fragment from artifi- cially contaminated samples, while neither false positive nor false negative results happens. The detecting limit of NASBA is 8.2pg/IxL nucleic acid in our test. The assay can amplify effectively on condition of 42~C , 90min. The detection of Shigella can be completed within 24 hours. Conclusion According to the advantage of specificity, sensitivity and short detection period, the NASBA assay is not only applicable to rapid detection of Shigella at fron- tier port, but also will have more extensive application in future with the development of technology.

关 键 词：志贺氏菌 NASBA检测 国境口岸 

分 类 号：R33[医药卫生—人体生理学]