应用DNA环介导恒温扩增技术快速检测空肠弯曲菌

doi:10.3864/j.issn.0578-1752.2012.13.020

中国农业科学 ›› 2012, Vol. 45 ›› Issue (13): 2751-2757.doi: 10.3864/j.issn.0578-1752.2012.13.020

应用DNA环介导恒温扩增技术快速检测空肠弯曲菌

徐义刚, 李丹丹, 田长永, 刘忠梅, 李苏龙, 张国财

1.黑龙江出入境检验检疫局检验检疫技术中心，哈尔滨 150001
2.东北林业大学，哈尔滨 150040
3.海南出入境检验检疫局检验检疫技术中心，海口 570125
4.辽宁农业职业技术学院，辽宁营口 115009

收稿日期:2011-07-29 出版日期:2012-07-01 发布日期:2012-06-01
通讯作者: 通信作者张国财，Tel：0451-82191756；E-mail：zhang640308@126.com
作者简介:徐义刚，E-mail：yigangxu_china@yahoo.com.cn
基金资助:
国家质检总局科技计划项目（2008IK162）、黑龙江省博士后基金（LBH-Z09）

Rapid Detection of Campylobacter jejuni Using Loop-Mediated Isothermal Amplification Assay

XU Yi-Gang, LI Dan-Dan, TIAN Chang-Yong, LIU Zhong-Mei, LI Su-Long, ZHANG Guo-Cai

1.黑龙江出入境检验检疫局检验检疫技术中心，哈尔滨 150001
2.东北林业大学，哈尔滨 150040
3.海南出入境检验检疫局检验检疫技术中心，海口 570125
4.辽宁农业职业技术学院，辽宁营口 115009

Received:2011-07-29 Online:2012-07-01 Published:2012-06-01

摘要/Abstract

摘要： 【目的】空肠弯曲菌是一种重要的人畜共患病致病菌，主要经被污染的水源、乳制品、生禽肉进行传播，建立快速、准确的诊断方法是防控空肠弯曲菌感染的重要措施。【方法】本文引入一种新型的核酸检测技术——DNA环介导恒温扩增 (loop-mediated isothermal amplification, LAMP)，以空肠弯曲菌ORF-C序列为检测靶基因，建立了空肠弯曲菌LAMP快速检测方法。【结果】利用本LAMP方法对21种病原菌进行检测，仅空肠弯曲菌为阳性结果，说明该方法具有高度的特异性。该LAMP方法对空肠弯曲菌纯培养菌的检测灵敏度达6CFU/反应管，对污染材料中空肠弯曲菌的检测灵敏度为9CFU/反应管。【结论】本研究建立的空肠弯曲菌LAMP检测方法操作简便、检测快速，具有良好的实用性。

关键词: 空肠弯曲菌, 环介导恒温扩增, 快速检测

Abstract: 【Objective】Campylobacter jejuni is a major zoonotic pathogen, mainly initiates its transmission with contaminated water, milk products and raw poultry. Thus, the development of a rapid and accurate diagnostic method is an important measure to prevent and control C. jejuni.【Method】Loop-mediated isothermal amplification (LAMP) assay was introduced to develop a rapid method for the detection of C. jejuni using the ORF-C sequence of C. jejuni as the target gene.【Result】Twenty-one bacterial strains were used to evaluate the specificity of the LAMP method. The result showed that only C. jejuni was positive and the other strains were negative in LAMP, indicating that the LAMP method was highly specific to the target pathogen with the sensitivity of 5CFU per test tube for C. jejuni cultivated and 9 CFU per test tube for its contaminated materials.【Conclusion】The LAMP method developed in this work is a sensitive, rapid and simple tool for the detection of C. jejuni.

Key words: Campylobacter jejuni, LAMP, rapid detection

徐义刚, 李丹丹, 田长永, 刘忠梅, 李苏龙, 张国财. 应用DNA环介导恒温扩增技术快速检测空肠弯曲菌[J]. 中国农业科学, 2012, 45(13): 2751-2757.

XU Yi-Gang, LI Dan-Dan, TIAN Chang-Yong, LIU Zhong-Mei, LI Su-Long, ZHANG Guo-Cai. Rapid Detection of Campylobacter jejuni Using Loop-Mediated Isothermal Amplification Assay[J]. Scientia Agricultura Sinica, 2012, 45(13): 2751-2757.

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应用DNA环介导恒温扩增技术快速检测空肠弯曲菌

Rapid Detection of Campylobacter jejuni Using Loop-Mediated Isothermal Amplification Assay

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