A novel Fusion-5 Filter based Micro-chip: A highly efficient, On-chip DNA extraction and On-chip PCR amplification for rapid detection

Abstract

A novel, sensitive, portable method for detecting DNA from various human samples is most desirable. This study aims to develop a microdevice for on-chip DNA extraction and detection by PCR from various forensic samples. A microchip constructed by sandwiching a fusion-5 filter disc between PMMA layers was characterized using SEM, FTIR, and XRD. DNA capture efficiency of the microchip from human samples was quantified by Real-Time PCR. PCR products were evaluated off-chip by DNA sequencing (STR Typing). On-site detection was performed by visualizing the DNA amplicons on Fusion-5 filter paper under UV light after EtBr staining. Among all concentrations, 5% by weight PMMA membrane was found most suitable for PMMA-Fusion-5 filter disc fluidic Microchip, the best smooth cross-section morphology by SEM, strong absorption vibrations at corresponding wavelengths by FTIR, increased amorphous phase by XRD were confirmed. Using this microdevice, DNA extraction from human whole blood was, without any leakage, fast (≤7 minutes), most efficient (highest, Ct=27.22) as evaluated by real-time PCR, needs just 2µl blood sample as shown by a typical, balanced STR profile. The microdevice designed for on-chip DNA extraction has excellent potential for rapid, on-site DNA detection from various samples.

URN:NBN:sciencein.cbl.2024.v11.665