Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, PR China.
Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, PR China.
Biosens Bioelectron. 2015 Dec 15;74:628-36. doi: 10.1016/j.bios.2015.07.019. Epub 2015 Jul 14.
Foodborne illnesses have always been a serious problem that threats public health, so it is necessary to develop a method that can detect the pathogens rapidly and sensitively. In this study, we designed a magnetic controlled microfluidic device which integrated the dynamic magnetophoretic separation and stationary magnetic trap together for sensitive and selective detection of Salmonella typhimurium (S. typhimurium). Coupled with immunomagnetic nanospheres (IMNs), this device could separate and enrich the target pathogens and realize the sensitive detection of target pathogens on chip. Based on the principle of sandwich immunoassays, the trapped target pathogens identified by streptavidin modified QDs (SA-QDs) were detected under an inverted fluorescence microscopy. A linear range was exhibited at the concentration from 1.0×10(4) to 1.0×10(6) colony-forming units/mL (CFU/mL), the limit of detection (LOD) was as low as 5.4×10(3) CFU/mL in milk (considering the sample volume, the absolute detection limit corresponded to 540C FU). Compared with the device with stationary magnetic trap alone, the integrated device enhanced anti-interference ability and increased detection sensitivity through dynamic magnetophoretic separation, and made the detection in complex samples more accurate. In addition, it had excellent specificity and good reproducibility. The developed system provides a rapid, sensitive and accurate approach to detect pathogens in practice samples.
食源性疾病一直是威胁公众健康的严重问题,因此有必要开发一种能够快速、灵敏地检测病原体的方法。本研究设计了一种磁控微流控装置,将动态磁泳分离和固定磁阱集成在一起,用于灵敏和选择性检测鼠伤寒沙门氏菌(S. typhimurium)。该装置与免疫磁纳米球(IMNs)结合,可分离和富集目标病原体,并在芯片上实现目标病原体的灵敏检测。基于夹心免疫分析原理,通过链霉亲和素修饰的量子点(SA-QDs)对捕获的目标病原体进行检测,在 1.0×10(4)至 1.0×10(6) 菌落形成单位/毫升(CFU/mL)的浓度范围内呈现线性范围,在牛奶中的检测限(LOD)低至 5.4×10(3) CFU/mL(考虑到样品体积,绝对检测限相当于 540C FU)。与仅具有固定磁阱的装置相比,集成装置通过动态磁泳分离增强了抗干扰能力并提高了检测灵敏度,使复杂样品中的检测更加准确。此外,该装置还具有出色的特异性和良好的重现性。所开发的系统为实际样品中病原体的检测提供了一种快速、灵敏和准确的方法。