Hand-held syringe as a portable plastic pump for on-chip continuous-flow PCR: miniaturization of sample injection device

Abstract

On-chip continuous-flow polymerase chain reactions (PCRs) generally require peripheral apparatus such as a pump for injecting a sample liquid into the fluidic channel. This makes the overall instrumentation bulky, limiting integration. In this study, we propose a new scheme for injecting a sample employing a hand-held syringe as a portable plastic pump, and apply it to an on-chip continuous-flow PCR. In the proposed injection scheme, sample actuation was realized inside a highly gas-permeable and blunt-ended fluidic conduit connected to a hand-held plastic syringe filled with compressed air. In this system, the degree of air diffusion via the walls of the gas-permeable conduit becomes greater in the anterior (closer to the outlet) end of the sample plug than the posterior (closer to the inlet) end, because a relatively larger quantity of air is retained inside the syringe at the posterior end of the sample plug. This creates a pressure gradient at the inlet and outlet of the fluidic conduit and propels the sample forward toward the outlet. Preliminary experiments were performed for the quantitative analyses and evaluation of the proposed sample injection scheme using gas-permeable silicone tubes. As practical applications, a 230 bp gene fragment from a plasmid vector and the first 282 bp of the interferon-beta (IFN-β) promoter from a human genomic DNA were successfully amplified on a microdevice coupled with a hand-held syringe as a portable sample actuation device, greatly enhancing device portability for on-site analyses.