多重RPA-LFD技术快速检测磺胺类耐药基因研究

Rapid detection of sulfonamide resistance genes and by multiplex RPA-LFD

目的:畜禽养殖业普遍存在抗生素过量使用甚至滥用的现象,导致动物源细菌耐药性问题越来越严重。磺胺类药物是应用最广泛的抗生素之一,对磺胺类耐药基因进行快速检测,有利于控制动物源耐药细菌的传播。方法:利用重组酶聚合酶扩增技术(RPA)在常温下快速扩增痕量核酸的特点,结合横向流动试纸条(LFD),对动物源大肠杆菌磺胺耐药基因 sul1,sul2和sul3进行同步快速检测,以实现现场可视化快速检测。结果:磺胺耐药基因sul1、sul2、sul3 在动物源大肠杆菌中的检出率分别为36.36%(84/231)、54.54%(126/231)和68.39%(158/231),检测限可达到10 3copies/μL,且无交叉反应。结论:此方法结果简单易读,可为磺胺类耐药基因现场可视化快速检测提供技术支撑。

Aims: Antibiotics abuse in the livestock and poultry farming industry leads to a serious problem of drug resistance to zoonotic bacteria. Sulfa drug is one of the most widely used antibiotics. Rapid detecting of sulfonamide resistance genes is helpful to control the spread of the zoonotic bacterial drug resistance. Methods: The recombinase polymerase amplification can rapidly amplify trace nucleic acids at room temperature. The lateral flow dipsticks can realize visual detection in field rapidly. In this paper, the multiple RPA-LFD technique was used to detect the sulfonamine-resistant genes sul1, sul 2 and sul3 in zoonotic Escherichia coli simultaneously and rapidly. Results: The detection rates of sulfonamine-resistant genes sul1, sul 2 and sul3 in zoonotic Escherichia coli were 36.36%(84/231), 54.54%(126/231), and 68.39%(158/231) respectively. The detection limit could reach 10 3copies;and there was no cross reaction. Conclusions: The method is simple and can provide technical support for the visual detection of sulfonamides-resistant genes rapidly in field.