An integrated rotary microfluidic system with DNA extraction, loop-mediated isothermal amplification, and lateral flow strip based detection for point-of-care pathogen diagnostics

다국어 초록 (Multilingual Abstract)

<P><B>Abstract</B></P> <P>Point-of-care (POC) molecular diagnostics plays a pivotal role for the prevention and treatment of infectious diseases. In spite of recent advancement in microfluidic based POC devices, there are still rooms for development to realize rapid, automatic and cost-effective sample-to-result genetic analysis. In this study, we propose an integrated rotary microfluidic system that is capable of performing glass microbead based DNA extraction, loop mediated isothermal amplification (LAMP), and colorimetric lateral flow strip based detection in a sequential manner with an optimized microfluidic design and a rotational speed control. Rotation direction-dependent coriolis force and siphon valving structures enable us to perform the fluidic control and metering, and the use of the lateral flow strip as a detection method renders all the analytical processes for nucleic acid test simplified and integrated without the need of expensive instruments or human intervention. As a proof of concept for point-of-care DNA diagnostics, we identified the food-borne bacterial pathogen which was contaminated in water or milk. Not only monoplex <I>Salmonella Typhimurium</I> but also multiplex <I>Salmonella Typhimurium</I> and <I>Vibrio parahaemolyticus</I> were analysed on the integrated rotary genetic analysis microsystem with a limit of detection of 50 CFU in 80min. In addition, three multiple samples were simultaneously analysed on a single device. The sample-to-result capability of the proposed microdevice provides great usefulness in the fields of clinical diagnostics, food safety and environment monitoring.</P> <P><B>Highlights</B></P> <P> <UL> <LI> An integrated rotary microsystem for point-of-care DNA diagnostics was demonstrated. </LI> <LI> The extraction, amplification and detection of DNA were integrated in a single disc. </LI> <LI> Food borne pathogens were successfully identified on the microdevice with a RPM control. </LI> <LI> The sample-to-result capability of the proposed microdevice would be adequate in early diagnosis. </LI> </UL> </P>