A portable microreactor with minimal accessories for polymerase chain reaction: application to the determination of foodborne pathogens

摘要

The authors describe a microreactor for performing a multiplexed polymerase chain reaction (PCR) which is operated with minimal accessories such as a single heater for gene amplification and a handheld syringe for sample actuation. It was fabricated by wrapping a polytetrafluoroethylene (PTFE) tube around a rectangular poly(dimethylsiloxane) (PDMS) block having a predetermined thickness. The resulting portable microreactor was applied to PCR using a single heater because the rectangular PDMS block spatially segregated the upper and lower arrays of the PTFE tube. This warrants the adjustment of distinct temperatures inside the upper and lower tubes. A hand-held plastic syringe was connected to the inlet of the PTFE tube and used as a portable pump to achieve a homogeneous transport of a sample liquid inside the microreactor. The use of gas-impermeable PTFE prevents the formation of bubbles during thermal cycling. The microreactor was successfully applied to the amplification of typical DNA fragments of three foodborne pathogens in less than 30 min. In our perception, this method paves the way to the construction of a truly portable PCR chip that is applicable to rapid clinical diagnosis and the detection of foodborne pathogens.