LAMP 技术在食源性病原微生物检测中的应用.docVIP

LAMP技术在食源性病原微生物检测中的应用 陈传1，凌霄2，郭震1，邵筱1，张如胜3，魏泉德3* （1.广东医学院，广东 湛江 524023；2.广东医学院第二附属医院，广东 湛江 524023；3.珠海市疾病预防与控制中心，广东 珠海519000） 目的 依据环介导恒等温扩增技术(LAMP)原理，建立一种快速、灵敏、特异、有效、适用于现场、战时野外及基层的食物中毒常见菌的检测体系。方法 本实验针对食物中毒常见的细菌的目的基因设计引物，并以此套引物建立基于LAMP技术的病原微生物检测方法。结果 灵敏性及特异性试验证明，该方法灵敏度高、特异性好，LAMP法最低检出限为10-Real-Time LAMP最低检出限为10-普通PCR方法；全部反应可在1h内完成；反应结果可通过肉眼观测直接判定结果。结论 初步建立了LAMP快速检测常见食物中毒菌的方法，该法能在简陋的环境和不需要特殊仪器设备的条件下简便、快速、特异、敏感地完成检测任务。为现场、战时野外或条件较差的实验室进行病原微生物的快速检测提供了很好的选择。 食源性微生物；LAMP技术；；Real-Time 中图分类号：R378.3 文献标识码：A 文章编号： The application of LAMP technology in detection for forborne pathogenic microorganisms CHEN Chuan1, LING Xiao2, GUO Zhen1, SHAO Xiao1, ZHANG Rusheng3, WEI Quande3* (1. Guangdong Medical College, Zhanjiang 524023, Guangdong, P. R. China; 2. 2nd Affiliated Hospital of Guangdong Medical College, Zhanjiang 524023, Guangdong, P. R. China; 3. Zhuhai Center for Disease Control Prevention, Zhuhai 519000, Guangdong, P. R. China) 【Abstracts】 Objective Based on the principle of the loop-mediated constant isothermal amplification (LAMP), to establish a rapid, sensitive, specific, effective and suitable for on-site, wartime field and grass-roots common food poisoning bacteria detection system. Method This experiment target gene primers were designed for common food poisoning bacteria, and thus sets the primers create a pathogenic microorganism diagnosis technology based on LAMP method. Results Sensitivity and specificity of experiments show that the method had high sensitivity and specificity. LAMP minimum sensitivity detection limit was 10-5, while the lowest detection limit of the Real-Time LAMP was 10-7, more than two orders of magnitude higher than conventional PCR methods. All the reaction can be completed in 1h , and reaction results can be directly judged by the naked eye observation. Conclusion Initially established based on the LAMP method for rapid detection of common food poisoning bacteria, the law under rough conditions and does not require sp