Extraction of genomic DNA and detection of single nucleotide polymorphism genotyping utilizing an integrated magnetic bead-based microfluidic platform

Abstract

This study presents a new magnetic bead-based microfluidic platform, which integrates three major modules for rapid leukocytes purification, genomic DNA (gDNA) extraction and fast analysis of genetic gene. By utilizing microfluidic technologies and magnetic beads conjugated with CD 15/45 antibodies, leukocytes in a human whole blood sample can be first purified and concentrated, followed by extraction of gDNA utilizing surface-charge switchable, DNA-specific, magnetic beads in the lysis solution. Then, specific genes associated with genetic diseases can be amplified by an on-chip polymerase chain reaction (PCR) process automatically. The whole pretreatment process including the leukocytes purification and gDNA extraction can be performed in an automatic fashion with the incorporation of the built bio-separators consisting of microcoils array within less than 20 min. The detection of single nucleotide polymorphism (SNP) genotyping of methylenetetra-hydrofolate reductase (MTHFR) C677T region associated with an increased risk of genetic diseases was further performed to demonstrate the capability of the proposed system. The extracted gDNA can be transported into a micro PCR chamber for on-chip fast nucleic acid amplification of detection genes with minimum human intervention. Hence, the developed system may provide a powerful automated platform for pretreatment of human leukocytes, gDNA extraction and fast analysis of genetic gene.

Polymeric-Based In Vitro Diagnostic Devices

“…Several mixer designs have been successfully incorporated into IVD devices. Membrane-based micromixers use air to expand and compress a series of chambers; they create creating mixing in fluidic channels as part of a platform for DNA extraction, leukocyte purification, and Re = ρUL µ genotyping from whole blood [184]. Air is also used to actuate multilayer PDMS chambers for mixing in urine analysis devices [185].…”

“…In conventional PCR diagnostics, sample preparation steps such as DNA extraction are time-consuming procedures that require a commercially available kit and experienced technical skill. Recent developments have overcome these barriers by incorporating integrated sample purification steps for on-chip DNA extraction [184,207,208]. Integrated LOC platforms use magnetic beads to purify and concentrate leukocytes prior to DNA extraction in lysis solution; this is followed by PCR amplification [184,207].…”

Immunomagnetic nanoparticle-based assays for detection of biomarkers

“…63 (B) Pillar-shaped magnets are used for automated transportation, washing, and detection of DNA biomarkers. 51 working principle of the magnetic separator (B); a, metal plate with the reaction tubes containing magnetic particles suspended in buffer solution; b, and c, for the washing steps, the metal plate with the magnets can be moved from one side of the tube to the other; d, carriers for sample lysates; e, carriers for disposable tips; f, waste container; g, magnetic separator; h, carriers for DNA eluates; i, barcode reader, j, liquid handling arm and k, computer.…”

3D-Printed Microfluidic Platform Enabling Bacterial Preconcentration and DNA Purification for Molecular Detection of Pathogens in Blood

“…Recently, efforts have been made to extract and isolate target DNA using either MSBs or IMS in μFDs [7-9, 15,16]. However, conventional μFDs [13][14][15][16] are typically not suitable for processing samples larger than 1 mL due to the small dimensions (~ 1 mm) of their microchannels. In addition, their fabrication requires multiple layers and several bonding steps, making them difficult to be massproduced.…”