Integrated, DC voltage-driven nucleic acid diagnostic platform for real sample analysis: Detection of oral cancer

Abstract

We present an integrated and low-cost microfluidic platform capable of extraction of nucleic acids from real biological samples. We demonstrate the application of this platform in pathogen detection and cancer screening. The integrated platform consists of three units including a pretreatment unit for separation of nucleic acids from lysates, a preconcentration unit for concentration of isolated nucleic acids and a sensing unit localized at a designated position on the chip for specific detection of the target nucleic acid. The platform is based on various electrokinetic phenomena exhibited by ion exchange membranes in a DC electrical field that allow them to serve as molecular filters, analyte preconcentrators and sensors. In this manuscript, we describe each unit of the integrated chip separately and show specific detection of a microRNA (miRNA 146a) biomarker associated with oral cancer as a proof-of-concept experiment. This platform technology can easily be extended to other targets of interest by optimizing the properties of the ion exchange membranes and the specific probes functionalized onto the sensors.

High-flux ionic diodes, ionic transistors and ionic amplifiers based on external ion concentration polarization by an ion exchange membrane: a new scalable ionic circuit platform

“…The AEM is embedded in a circular polyurethane resin pellet and fabricated using a specific molding protocol. 25 A PMMA tubing glued to the resin pellet serves as the “G” terminal reservoir. The reservoirs for the three electrical terminals “D”, “S” and “B” are cut pipette tips and are separated from the channels by filter papers and a layer of 1% agarose gel to prevent possible bubble entry into the channels.…”

Sensor-Integrated Microfluidic Approaches for Liquid Biopsies Applications in Early Detection of Cancer

“…As a proof of concept, different biological samples were analyzed in MERCA system. Another integrated microfluidic technology was based on an ionic membrane modular biochip platform developed by Slouka et al, 2015 [124] (Figure 5C). With this modern technique, it is possible to rapidly measure (30-60 min) a maximum of three different miRNAs, stating an LOD of 106 copies with high selectivity and sensitivity, using raw cell media or cell lysate as the substrate.…”

“…The final target extraction is carried out by a probe-functioned membrane that enhances selectivity and allows the effective quantification of the target probe. [124].…”

Current Technologies and Recent Developments for Screening of HPV-Associated Cervical and Oropharyngeal Cancers

“…In contrast, Slouka et al and Senapati et al developed a simple, single-channel integrated diagnostic platform as shown in Figure 2 , consisting of a pretreatment unit for extraction of nucleic acid biomarkers, a preconcentration unit to concentrate the extracted nucleic acids (reduces assay time by minimizing diffusion limitation) and a nanomembrane-based biosensing unit for selective detection of target nucleic acids [ 195 , 196 ]. The team utilized the inherent charge of nucleic acid molecules and ion-exchange membranes to develop an integrated multiplexed diagnostics platform capable of processing real biological samples and detecting pathogens in 45 min, often without PCR amplification for overexpressed miR targets [ 196 ]. While further validation of this platform using biological and clinical samples is ongoing, the unique features of this system are highlighted below.…”