Current methods for diagnosis of human coronaviruses: pros and cons.

The current global fight against coronavirus disease (COVID-19) to flatten the transmission curve is put forth by the World Health Organization (WHO) as there is no immediate diagnosis or cure for COVID-19 so far. In order to stop the spread, researchers worldwide are working around the clock aiming to develop reliable tools for early diagnosis of severe acute respiratory syndrome (SARS-CoV-2) understanding the infection path and mechanisms. Currently, nucleic acid-based molecular diagnosis (real-time reverse transcription polymerase chain reaction (RT-PCR) test) is considered the gold standard for early diagnosis of SARS-CoV-2.

Cryoprotectant-Free Freezing of Cells Using Liquid Marbles Filled with Hydrogel.

Cryopreservation without cryoprotectant remains a significant challenge for the re-establishment of cell culture after freeze-thaw. Thus, finding an alternative and a simple cryopreservation method is necessary. Liquid marble (LM)-based digital microfluidics is a promising approach for cryoprotectant-free cryopreservation.

A bisulfite treatment and PCR-free global DNA methylation detection method using electrochemical enzymatic signal engagement.

In this paper we report on a bisulfite treatment and PCR amplification-free method for sensitive and selective quantifying of global DNA methylation. Our method utilizes a three-step strategy that involves (i) initial isolation and denaturation of global DNA using the standard isolation protocol and direct adsorption onto a bare gold electrode via gold-DNA affinity interaction, (ii) selective interrogation of methylation sites in adsorbed DNA via methylation-specific 5mC antibody, and (iii) subsequent signal enhancement using an electrochemical-enzymatic redox cycling reaction. In the redox cycling reaction, glucose oxidase (GO) is used as an enzyme label, glucose as a substrate and ruthenium complex as a redox mediator.

DNA methylation detection: recent developments in bisulfite free electrochemical and optical approaches.

DNA methylation is one of the significant epigenetic modifications involved in mammalian development as well as in the initiation and progression of various diseases like cancer. Over the past few decades, an enormous amount of research has been carried out for the quantification of DNA methylation in the mammalian genome. Earlier, most of these methodologies used bisulfite treatment.

Porous nanozymes: the peroxidase-mimetic activity of mesoporous iron oxide for the colorimetric and electrochemical detection of global DNA methylation.

Nanomaterials (nanozymes) with peroxidase-mimetic activity have been widely used in biosensing platforms as low-cost, relatively stable and prevailing alternatives to natural enzymes. Herein, we report on the synthesis and application of the peroxidase-mimetic activity of mesoporous iron oxide (MIO) for the detection of global DNA methylation in colorectal cancer cell lines. The target DNA was extracted and denatured to get ssDNA followed by direct adsorption onto the surface of a bare screen-printed gold electrode (SPGE).

Mesoporous Iron Oxide Synthesized Using Poly(styrene-b-acrylic acid-b-ethylene glycol) Block Copolymer Micelles as Templates for Colorimetric and Electrochemical Detection of Glucose.

Herein, we report the soft-templated preparation of mesoporous iron oxide using an asymmetric poly(styrene-b-acrylic acid-b-ethylene glycol) (PS-b-PAA-b-PEG) triblock copolymer. This polymer forms a micelle consisting of a PS core, a PAA shell, and a PEG corona in aqueous solutions, which can serve as a soft template. The mesoporous iron oxide obtained at an optimized calcination temperature of 400 °C exhibited an average pore diameter of 39 nm, with large specific surface area and pore volume of 86.

Quantification of gene-specific DNA methylation in oesophageal cancer via electrochemistry.

Development of simple and inexpensive method for the analysis of gene-specific DNA methylation is important for the diagnosis and prognosis of patients with cancer. Herein, we report a relatively simple and inexpensive electrochemical method for the sensitive and selective detection of gene-specific DNA methylation in oesophageal cancer. The underlying principle of the method relies on the affinity interaction between DNA bases and unmodified gold electrode.

Colorimetric and electrochemical quantification of global DNA methylation using a methyl cytosine-specific antibody.

We report a simple colorimetric (naked-eye) and electrochemical method for the rapid, sensitive and specific quantification of global methylation levels using only 25 ng of input DNA. Our approach utilises a three-step strategy; (i) initial adsorption of the extracted, purified and denatured bisulfite-treated DNA on a screen-printed gold electrode (SPE-Au), (ii) immuno-recognition of methylated DNA using a horseradish peroxidase (HRP)-conjugated methylcytosine (HRP-5mC) antibody and (iii) subsequent colorimetric detection by the enzymatic oxidation of 3,3',5,5'-tetramethylbenzidin (TMB)/HO which generated a blue-coloured product in the presence of methylated DNA and HRP-5mC immunocomplex. As TMB is electroactive, it also produces detectable amperometric current at +150 mV versus a Ag pseudo-reference electrode (electrochemical detection).