Por favor, use este identificador para citar o enlazar este ítem: https://hdl.handle.net/20.500.12815/193
Título : 3D-printed modular microfluidic device enabling preconcentrating bacteria and purifying bacterial DNA in blood for improving the sensitivity of molecular diagnostics
Autor: Abafogi, Abdurhaman Teyib
Kim, Jaewon
Lee, Jinyeop
Mohammed, Merem Omer
van Noort, Danny
Park, Sungsu
Fecha de publicación : 21-feb-2020
Resumen : Molecular diagnostics for sepsis is still a challenge due to the presence of compounds that interfere with gene amplification and bacteria at concentrations lower than the limit of detection (LOD). Here, we report on the development of a 3D printed modular microfluidic device (3DpmµFD) that preconcentrates bacteria of interest in whole blood and purifies their genomic DNA (gDNA). It is composed of a W-shaped microchannel and a conical microchamber. Bacteria of interest are magnetically captured from blood in the device with antibody conjugated magnetic nanoparticles (Ab-MNPs) at 5 mL/min in the W-shaped microchannel, while purified gDNA of the preconcentrated bacteria is obtained with magnetic silica beads (MSBs) at 2 mL/min in the conical microchamber. The conical microchamber was designed to be connected to the microchannel after the capturing process using a 3D-printed rotary valve to minimize the exposure of the MSBs to interfering compounds in blood. The pretreatment process of spiked blood (2.5 mL) can be effectively completed within about 50 min. With the 3DpmµFD, the LOD for the target microorganism Escherichia coli O157:H7 measured by both polymerase chain reaction (PCR) with electrophoresis and quantitative PCR was 10 colony forming unit (CFU) per mL of whole blood. The results suggest that our method lowers the LOD of molecular diagnostics for pathogens in blood by providing bacterial gDNA at high purity and concentration.
Palabras clave : Molecular diagnostics
Microfluidic device
Bacterial preconcentration
DNA purification
Pathogen
Editorial : MDPI
URI: https://hdl.handle.net/20.500.12815/193
DOI : https://doi.org/10.3390/s20041202
Aparece en las colecciones: Artículos Científicos

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