基于环介导等温扩增的离心式微流控芯片检测3种致病菌

The detection of three pathogenic bacteria by centrifugal microfluidic chip based on LAMP methodology

采用微流控技术可以将食源性致病菌样品的制备、分离、检测等基本单元集成到仅有几平方厘米的芯片上,与分子检测技术相结合,能够满足对食源性致病菌快速现场检测的需要。基于环介导等温扩增技术(loop-mediated isothermal amplification, LAMP),设计制作出一种用于食源性致病菌快速检测的离心式微流控芯片。通过离心式微流控核酸等温扩增装置实现微流控芯片上的流体控制、等温扩增及可视化检测,可1次进行5种样品的3种食源性致病菌的检测。结果显示,基于LAMP的离心式微流控方法用于检测大肠杆菌、沙门氏菌和单增李斯特氏菌,能够在60 min内完成芯片的可视化检测,并且检测结果具有良好的特异性及敏感性,检测限为10^(3)copies/μL。将微流控技术用于食源性致病菌的核酸提取、等温扩增及检测,具有集成化、易于携带和快速分析等优势,在食品安全快速检测中具有应用的潜力。

Microfluidic technology can integrate basic units such as sample preparation, separation, and detection on a chip which is only a few square centimeters. It can be combined with molecular detection technology to meet the requirements of rapid on-site testing of food-borne pathogens. Based on the loop-mediated isothermal amplification technology, a centrifugal microfluidic chip for the rapid detection of food-borne pathogens was designed and manufactured. The chip included a sample reservoir, aliquoting chambers, reaction chambers, an injection hole, a vent, capillary valves and other structures. The centrifugal microfluidic nucleic acid isothermal amplification device realized fluid control, isothermal amplification and visual detection on the microfluidic chip, and could detect three food-borne pathogens in five samples at a time. The results showed that E. coli, Salmonella and Listeria monocytogenes could be detected with the loop-mediated isothermal amplification(LAMP)-based centrifugal microfluidic method within 60 minutes, and the results had good specificity and sensitivity. The lowest detection limit was 10^(3)copies/μL. Microfluidic technology is used for nucleic acid extraction, isothermal amplification and detection of food-borne pathogens. It has the advantages of integration, ease of carrying and rapid analysis, showing the potential of microfluidic technology in the rapid detection of food safety.