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

Biotechnology and Bioengineering
Quang Nghia PhamNae Yoon Lee

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.

References

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Citations

May 28, 2019·Journal of Nanobiotechnology·Monika RuzyckaIreneusz P Grudzinski
Aug 23, 2020·The Science of the Total Environment·Anna ToldràMònica Campàs

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Methods Mentioned

BETA
PCR
genotyping
fluorescence imaging
thermal treatment
electrophoresis
Fluorescence

Software Mentioned

ThermaCAM Researcher
ProgRes ® Capture Pro
Image Lab
Image Pro

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