Molecular test for COVID-19 diagnosis based on a colorimetric genomagnetic assay.

The world is in a long pandemic period caused by the SARS-CoV-2 virus and massive diagnostic tests to assist efforts to control the spread of the disease and also to avoid new coronavirus variants are still needed. Herein, we propose a simple and accurate saliva-based colorimetric test for the diagnosis of COVID-19. Magnetic beads (MBs) modified with a sequence of single-strand DNA (ssDNA) complementary to the N gene of the SARS-CoV-2 RNA were developed and used for magnetic capture and separation from a complex saliva sample. A second biotinylated ssDNA sequence was applied, and the colorimetric detection was carried out by adding streptavidin-horseradish peroxidase conjugate, H2O2, and tetramethylbenzidine (TMB) as chromogenic substrate. The test does not require viral RNA isolation, transcription, or amplification steps and can be performed at room temperature. The molecular assay test can be run using 96-well microplates, allowing the diagnosis of a large number of samples in 90 min. A simple support for magnets was designed and constructed using a 3D printer that allows the magnetic separations directly in the 96-well microplate. The colorimetric test showed an excellent ability to discriminate between healthy individuals and patients infected with SARS-CoV-2, with 92% and 100% of clinical sensitivity and specificity, respectively. This performance was similar to that achieved using the gold standard RT-PCR technique. The proposed genomagnetic assay offers an opportunity to greatly increase population testing, contribute to controlling the spread of the virus, and improve health equity in testing for COVID-19.

Enzymatic biosensors based on ingá-cipó peroxidase immobilised on sepiolite for TBHQ quantification.

Sepiolite clay mineral was used as a support for the immobilisation of the peroxidase enzyme from ingá-cipó (Inga edulis Mart.) and was used with graphite powder, multi-walled carbon nanotubes (CNTs), mineral oil, and nafion 0.5% (v/v) in the development of a new biosensor for the determination of the antioxidant tert-butylhydroquinone (TBHQ) by square-wave voltammetry (SWV). For the optimisation and application of the biosensor, several parameters were investigated to determine the optimum experimental conditions using SWV. The best performance was obtained using a 0.1 mol L(-1) phosphate buffer solution (pH 7.0), 4.0 × 10(-4) mol L(-1) hydrogen peroxide, a frequency of 50 Hz, a pulse amplitude of 60 mV, and a scan increment of 6 mV. The biosensor showed good repeatability and reproducibility and remained stable for a period of 20 weeks. The analytical curve revealed a linear response range of 1.65 to 9.82 mg L(-1) (r = 0.994) with detection and quantification limits of 0.41 and 1.25 mg L(-1). A recovery study of TBHQ in salad dressing samples yielded values from 99.6-104.8%. The proposed biosensor was successfully used for the determination of TBHQ in commercial salad dressing samples, giving a relative error of 5.4% in relation to the comparative method (chromatographic).