3D-Printed Modular Microfluidic Device Enabling Preconcentrating Bacteria and Purifying Bacterial DNA in Blood for Improving the Sensitivity of Molecular Diagnostics
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...
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doaj-8d443d8185364fda86506d1a0230e6a82020-11-25T00:31:11ZengMDPI AGSensors1424-82202020-02-01204120210.3390/s20041202s200412023D-Printed Modular Microfluidic Device Enabling Preconcentrating Bacteria and Purifying Bacterial DNA in Blood for Improving the Sensitivity of Molecular DiagnosticsAbdurhaman Teyib Abafogi0Jaewon Kim1Jinyeop Lee2Merem Omer Mohammed3Danny van Noort4Sungsu Park5School of Mechanical Engineering, Sungkyunkwan University, Suwon 16419, KoreaSchool of Mechanical Engineering, Sungkyunkwan University, Suwon 16419, KoreaSchool of Mechanical Engineering, Sungkyunkwan University, Suwon 16419, KoreaDepartment of Biomedical Engineering, Sungkyunkwan University, Suwon 16419, KoreaDivision of Biotechnology, IFM, Linkoping University, 58183 Linkoping, SwedenSchool of Mechanical Engineering, Sungkyunkwan University, Suwon 16419, KoreaMolecular 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 <i>Escherichia coli</i> 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.https://www.mdpi.com/1424-8220/20/4/1202molecular diagnosticsmicrofluidic devicebacterial preconcentrationdna purificationpathogen |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Abdurhaman Teyib Abafogi Jaewon Kim Jinyeop Lee Merem Omer Mohammed Danny van Noort Sungsu Park |
spellingShingle |
Abdurhaman Teyib Abafogi Jaewon Kim Jinyeop Lee Merem Omer Mohammed Danny van Noort Sungsu Park 3D-Printed Modular Microfluidic Device Enabling Preconcentrating Bacteria and Purifying Bacterial DNA in Blood for Improving the Sensitivity of Molecular Diagnostics Sensors molecular diagnostics microfluidic device bacterial preconcentration dna purification pathogen |
author_facet |
Abdurhaman Teyib Abafogi Jaewon Kim Jinyeop Lee Merem Omer Mohammed Danny van Noort Sungsu Park |
author_sort |
Abdurhaman Teyib Abafogi |
title |
3D-Printed Modular Microfluidic Device Enabling Preconcentrating Bacteria and Purifying Bacterial DNA in Blood for Improving the Sensitivity of Molecular Diagnostics |
title_short |
3D-Printed Modular Microfluidic Device Enabling Preconcentrating Bacteria and Purifying Bacterial DNA in Blood for Improving the Sensitivity of Molecular Diagnostics |
title_full |
3D-Printed Modular Microfluidic Device Enabling Preconcentrating Bacteria and Purifying Bacterial DNA in Blood for Improving the Sensitivity of Molecular Diagnostics |
title_fullStr |
3D-Printed Modular Microfluidic Device Enabling Preconcentrating Bacteria and Purifying Bacterial DNA in Blood for Improving the Sensitivity of Molecular Diagnostics |
title_full_unstemmed |
3D-Printed Modular Microfluidic Device Enabling Preconcentrating Bacteria and Purifying Bacterial DNA in Blood for Improving the Sensitivity of Molecular Diagnostics |
title_sort |
3d-printed modular microfluidic device enabling preconcentrating bacteria and purifying bacterial dna in blood for improving the sensitivity of molecular diagnostics |
publisher |
MDPI AG |
series |
Sensors |
issn |
1424-8220 |
publishDate |
2020-02-01 |
description |
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 <i>Escherichia coli</i> 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. |
topic |
molecular diagnostics microfluidic device bacterial preconcentration dna purification pathogen |
url |
https://www.mdpi.com/1424-8220/20/4/1202 |
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