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石琰璟1， 王建广1，2， 房保海2， 张健2，
祝素珍2， 刘云国2，姜英辉2， 雷质文2*， 杨大伟2
(1．青岛科技大学 化工学院，山东 青岛 266042；2.山东出入境检验检疫局，山东 青岛 266002)
摘要: 以副溶血性弧菌的tlh基因为目的片段设计特异性引物，成功建立了恒温条件下快速检测副溶血性弧菌的依赖解旋酶DNA恒温扩增技术(HDA)的检测法，进行了特异性和灵敏度实验，并与普通PCR方法进行了比较。该HDA检测方法最低检测限为19.9 ng·mL-1，与普通PCR方法相当。副溶血性弧菌的HDA检测方法具有普通PCR的特异、灵敏等特点，并且对仪器要求更低，用普通水浴槽即可进行反应。
关键词: 副溶血性弧菌； 检测； 依赖解旋酶DNA恒温扩增技术
中图分类号: S 917.4 文献标志码： A
Detection of Vibrio Parahaemolyticus by Helicase-dependent Isothermal DNA Amplification Technology
SHI Yan-jing1， WANG Jian-guang1,2， FANG Bao-hai2, ZHANG Jian2,
ZHU Su-zhen2, LIU Yun-guo2， JIANG Ying-hui2, LEI Zhi-wen2, YANG Da-wei2
(1.College of Chemical Engineering, Qingdao University of Science and Technology，Qingdao 266042，China；
2.Shandong Entry-exit Inspection and Quarantine Bureau,Qingdao 266002,China)
Abstract: A new rapid method to detect Vibrio parahaemolyticus samples has been established on the basis of Helicase-dependent isothermal DNA amplification (HDA).A highly specific set of primers were synthesized to target the tlh gene of Vibrio parahaemolyticus. Specificity and sensitivity were tested by comparing with PCR method. The sensitivity was 19.9 ng·mL-1 which was similar to the result of PCR method. Detecting Vibrio parahaemolyticus with HDA is specific and sensitive as well as PCR method and has lower instrumental requirement.
Key words: Vibrio parahaemolyticus； detection； helicase-dependent isothermal DNA amplification
收稿日期: 2010-09-07
基金项目: 国家质检总局科研项目(2008IK140)；山东出入境检验检疫局科研项目(SK200821).
作者简介: 石琰璟(1974—)，女，副教授. *通信联系人.
