小体积水样中鼠伤寒沙门菌氨基磁珠富集的LAMP可视化检测方法  

Visualized LAMP Determination of Salmonella typhimurium in Small Volume Water Samples by Amine-Functionalized Magnetic Particle Concentration

作　　者：石雪梅 蒲荣春 郑江梅子 刘恋 阮佳 SHI Xuemei;PU Rongchun;ZHENG Jiangmeizi;LIU Lian;RUAN Jia(School of Public Health,Chengdu Medical College,Chengdu 621000,China;Nanchong Gaoping District Center for Disease Control and Prevention,Nanchong 637100,China)

机构地区：[1]成都医学院公共卫生学院,四川成都621000 [2]南充市高坪区疾病预防控制中心,四川南充637100

出　　处：《净水技术》2023年第3期189-197,共9页Water Purification Technology

基　　金：四川省自然科学基金项目(2022NSFSC0649);成都医学院教育教学改革项目(JG202120);四川省大学生创新创业训练计划项目(s201913705115)。

摘　　要：研究建立了小体积(1 mL)水样中鼠伤寒沙门菌氨基磁珠富集-环介导等温扩增(LAMP)可视化检测方法。研究对氨基磁珠富集水中鼠伤寒沙门菌的pH和时间进行优化,考察了氨基磁珠富集-LAMP可视化检测方法的特异性、抗干扰能力和灵敏度,并用所建立的方法对人工模拟水样和实际水样进行检测。在pH值=7.0的条件下,氨基磁珠与水样作用60 min,氨基磁珠对水中的鼠伤寒沙门菌的捕获效率接近100%。氨基磁珠富集-LAMP扩增体系特异性好,抗干扰能力强,对水中鼠伤寒沙门菌的检测灵敏度为2.39 CFU/mL,阳性结果可直接观察到反应管由橙黄色变成草绿色,而阴性体系不变色。将所建立的方法用于人工模拟水样,检测结果均为阳性,对成都市新都区河流或景观湖等水样检测,共检出1份鼠伤寒沙门菌阳性水样。本研究建立的氨基磁珠富集-LAMP方法能够实现小体积水样中鼠伤寒沙门菌高效、准确的可视化检测。A visualized determination method for the detection of Salmonella typhimurium in small volume water samples(1 mL)by amine-functionalized magnetic particle concentration-loop-mediated isothermal amplification(LAMP)was established in this study.In this research,the amine-functionalized magnetic particle concentration conditions,including the pH value and the enrichment time,were optimized.And the specificity,anti-interference performance and the sensitivity of the amine-functionalized magnetic particle concentration-LAMP method were investigated,respectively.What’s more,the established method was used to detect the Salmonella typhimurium in artificially contaminated water samples and actual water samples,either.Under the optimized conditions(pH value of the enrichment solution was 7.0,and enrichment time was 60 min),the recovery rate of the Salmonella typhimurium in water samples was close to 100%.The amine-functionalized magnetic particle concentration-LAMP method showed a good specificity and anti-interference performance.And the sensitivity of the method for Salmonella typhimurium in water samples was 2.39 CFU/mL.The reaction tube could be changed from orange to green for positive result,but it was still orange for the negative result.The results were all positive for the artificial contaminated water samples by the amine-functionalized magnetic particle concentration-LAMP method.There was one Salmonella typhimurium positive water sample when the river and landscape lake samples in Xindu District,Chengdu City were detected by the established method.The amine-functionalized magnetic particle concentration-LAMP method established in this research can visually detect the Salmonella typhimurium in small volume water samples effectively and accurately.

关 键 词：水环境 鼠伤寒沙门菌 富集 氨基磁珠 环介导等温扩增(LAMP) 

分 类 号：R123[医药卫生—环境卫生学]