Microdevice‐based solid‐phase polymerase chain reaction for rapid detection of pathogenic microorganisms

Abstract

We demonstrate the integration of DNA amplification and detection functionalities developed on a lab-on-a-chip microdevice utilizing solid-phase polymerase chain reaction (SP-PCR) for point-of-need (PON) DNA analyses. First, the polycarbonate microdevice was fabricated by thermal bonding to contain microchambers as reservoirs for performing SP-PCR. Next, the microchambers were subsequently modified with polyethyleneimine and glutaraldehyde for immobilizing amine-modified forward primers. During SP-PCR, the immobilized forward primers and freely diffusing fluorescence-labeled reverse primers cooperated to generate target amplicons, which remained covalently attached to the microchambers for the fluorescence detection. The SP-PCR microdevice was used for the direct identifications of two widely detected foodborne pathogens, namely Salmonella spp. and Staphylococcus aureus, and an alga causing harmful algal blooms annually in South Korea, Cochlodinium polykrikoides. The SP-PCR microdevice would be versatilely applied in PON testing as a universal platform for the fast identification of foodborne pathogens and environmentally threatening biogenic targets.

Microfluidics for studying metastatic patterns of lung cancer

“…The miniaturization of analytical and omics techniques has been previously predicted [1], however there is still a need of incorporation all these techniques into single microdevice. Progress in this field is noticeable since microdevices for DNA nanotechnology [231] and proteomic and metabolomic profiling [232] have been recently developed, however for maintaining reproducibly and repeatability of the results and eliminating cells contamination and errors related to material processing all techniques should be combined in a way of merging cell handling with analytical, biochemical and molecular determination.…”