5种口岸重要媒介动物传播病原体微流控芯片检测方法的建立

Establishment of microfluidic chip assay for rapid detection of five important vector-borne pathogens at port

目的建立口岸重点关注的鼠疫杆菌等5种经媒介动物传播病原体的微流控芯片检测方法,提升这5种重要病原体的快速检测能力。方法利用环介导等温扩增技术,将反应体系固化在同一块塑料芯片上,建立了同时检测鼠疫杆菌、伯氏疏螺旋体、土拉弗朗西斯菌、沙门菌、非结核分枝杆菌等5种重要病原体的微流控芯片检测方法。结果建立的微流控芯片检测方法特异性好,含有目的基因的阳性样本仅在芯片上相应病原体的反应槽出现显著扩增,而金黄色葡萄球菌等其他病原体反应槽未出现扩增,无交叉反应;沙门菌检测的敏感性为1×10^(2)拷贝/ml;检测方法具有良好的稳定性和可靠性。结论微流控芯片检测方法可为口岸快速筛查、排查疫情和快速处置提供重要的技术支持。

Objective To establish a microfluidic chip assay for rapid detection of five important vector-borne pathogens at port,and accelerate the rapid detection efficiency.Methods The reaction buffer system was solidified on the plastic chip by using loop mediated isothermal amplification(LAMP).A microfluidic chip detection method for simultaneously detecting five important pathogens including Yersinia pestis,Borrelia burgdorferi,Francisella tularensis,Salmonella and non-Tuberculous Mycobacterium was established.Results The results showed that the microfluidic chip assay had good specificity to these five pathogens and had no response to other pathogens such as Staphylcoccus aureus.The sensitivity of this method was 1×10^(2) copies/ml.The excellent stability of the microfluidic chip assay was further confirmed.Conclusion Microfluidic chip detection method provides important technical support for rapid screening,epidemic investigation and rapid disposal of imported infectious disease at ports.