Microfluidic Array Chip for Parallel Detection of Waterborne Bacteria

The polymerase chain reaction (PCR) is a robust technique used to make multiple copies of a segment of DNA. However, the available PCR platforms require elaborate and time-consuming operations or costly instruments, hindering their application. Herein, we introduce a sandwiched glass–polydimethylsiloxane (PDMS)–glass microchip containing an array of reactors for the real-time PCR-based detection of multiple waterborne bacteria. The PCR solution was loaded into the array of reactors in a single step utilising capillary filling, eliminating the need for pumps, valves, and liquid handling instruments. Issues of generating and trapping bubbles during the loading chip step were addressed by creating smooth internal reactor surfaces. Triton X-100 was used to enhance PCR compatibility in the chip by minimising the nonspecific adsorption of enzymes. A custom-made real-time PCR instrument was also fabricated to provide thermal cycling to the array chip. The microfluidic device was successfully demonstrated for microbial faecal source tracking (MST) in water

Microfluidic-Based Nucleic Acid Amplification Systems in Microbiology

Rapid, sensitive, and selective bacterial detection is a hot topic, because the progress in this research area has had a broad range of applications. Novel and innovative strategies for detection and identification of bacterial nucleic acids are important for practical applications. Microfluidics is an emerging technology that only requires small amounts of liquid samples. Microfluidic devices allow for rapid advances in microbiology, enabling access to methods of amplifying nucleic acid molecules and overcoming difficulties faced by conventional. In this review, we summarize the recent progress in microfluidics-based polymerase chain reaction devices for the detection of nucleic acid biomarkers. The paper also discusses the recent development of isothermal nucleic acid amplification and droplet-based microfluidics devices. We discuss recent microfluidic techniques for sample preparation prior to the amplification process

Microfluidic array chip for parallel detection of waterborne bacteria

The polymerase chain reaction (PCR) is a robust technique used to make multiple copies of a segment of DNA. However, the available PCR platforms require elaborate and time-consuming operations or costly instruments, hindering their application. Herein, we introduce a sandwiched glass-polydimethylsiloxane (PDMS)-glass microchip containing an array of reactors for the real-time PCR-based detection of multiple waterborne bacteria. The PCR solution was loaded into the array of reactors in a single step utilising capillary filling, eliminating the need for pumps, valves, and liquid handling instruments. Issues of generating and trapping bubbles during the loading chip step were addressed by creating smooth internal reactor surfaces. Triton X-100 was used to enhance PCR compatibility in the chip by minimising the nonspecific adsorption of enzymes. A custom-made real-time PCR instrument was also fabricated to provide thermal cycling to the array chip. The microfluidic device was successfully demonstrated for microbial faecal source tracking (MST) in water

Circulating tumor DNA and liquid biopsy: opportunities, challenges, and recent advances in detection technologies.

Opportunities and challenges in translational application of ctDNA along with recent developments in chip-based ctDNA detection technologies have been reviewed.</p

Graphene-Oxide-Loaded Superparamagnetic Iron Oxide Nanoparticles for Ultrasensitive Electrocatalytic Detection of MicroRNA

We report the electrocatalytic activity of a new class of superparamagnetic nanoparticles, graphene-oxide-loaded iron oxide (GO/IO hybrid material), towards the reduction of ruthenium hexaammine(III) chloride (Ru(NH3)6]3+, RuHex). Leveraging the electrocatalytic activity of the GO/IO hybrid material and the signal enhancement capacity of [Ru(NH3)6]3+/[Fe(CN)6]3 in an electrocatalytic cycle, an ultrasensitive and specific electrochemical sensor was developed for the detection of cancerrelated microRNA (miRNA). Using the direct affinity interaction between RNA and graphene oxide, magnetically isolated and purified target miRNA were directly adsorbed onto a screenprinted electrode modified with the GO/IO hybrid material. The detection was enabled by chronocoulometric (CC) readout of charge-compensating [Ru(NH3)6]3+ followed by an enhancement in CC charge display through the Ru(NH3)6]3+ /[Fe(CN)6]3 system. We demonstrate an excellent limit of detection of 1.0 fM by accurately detecting miR-21 in synthetic samples and showcase its clinical utility in ovarian cancer cell lines with high sensitivity (ten cells) and good reproducibility (%RSD= \u3c5%, for n=3)

Core-Shell Beads Made by Composite Liquid Marble Technology as A Versatile Microreactor for Polymerase Chain Reaction

Over the last three decades, the protocols and procedures of the DNA amplification technique, polymerase chain reaction (PCR), have been optimized and well developed. However, there have been no significant innovations in processes for sample dispersion for PCR that have reduced the amount of single-use or unrecyclable plastic waste produced. To address the issue of plastic waste, this paper reports the synthesis and successful use of a core-shell bead microreactor using photopolymerization of a composite liquid marble as a dispersion process. This platform uses the core-shell bead as a simple and effective sample dispersion medium that significantly reduces plastic waste generated compared to conventional PCR processes. Other improvements over conventional PCR processes of the novel dispersion platform include increasing the throughput capability, enhancing the performance and portability of the thermal cycler, and allowing for the contamination-free storage of samples after thermal cycling

Detection of FGFR2: FAM76A fusion gene in circulating tumor RNA based on catalytic signal amplification of graphene oxide-loaded magnetic nanoparticles

Circulating tumor nucleic acids (ctNAs) are promising biomarkers for minimally invasive cancer assessment. The FGFR2 : FAM76A fusion gene is one of the highly promising ovarian cancer biomarkers detectable in ctNAs. Herein, we introduce a new amplification-free electrochemical assay for the detection of FGFR2 : FAM76A fusion gene in ctNAs extracted from ovarian cancer patients. The assay relies on the electrocatalytic activity of a new class of superparamagnetic graphene-loaded iron oxide nanoparticles (GO-NPFeO). After isolation and purification, the target RNA was directly adsorbed onto the GO-NPFeO surface through graphene-RNA affinity interaction. The electrocatalytic signal was achieved by the reduction of surface-attached ruthenium hexaammine(III) chloride which was further amplified by using the ferricyanide redox system. Our assay depicted an excellent detection sensitivity down to 1.0 fM, high specificity and excellent reproducibility (% RSD