Microchip RT-PCR Detection of Nasopharyngeal SARS-CoV-2 Samples.

Fast, accurate, and reliable diagnostic tests are critical for controlling the spread of the coronavirus disease 2019 (COVID-19) associated with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. The current gold standard for testing is real-time PCR; however, during the current pandemic, supplies of testing kits and reagents have been limited. We report the validation of a rapid (30 minutes), user-friendly, and accurate microchip real-time PCR assay for detection of SARS-CoV-2 from nasopharyngeal swab RNA extracts.

A High-Throughput Screening Assay for NKCC1 Cotransporter Using Nonradioactive Rubidium Flux Technology.

A high-throughput screening (HTS) assay was developed for cotransporter, NKCC1, which is a potential target for the treatment of diverse disorders. This nonradioactive rubidium flux assay coupled with ion channel reader series provides a working screen for this target expressed in human embryonic kidney (HEK) cell line. An eightfold window of detection was achieved with the optimized assay.

The ninth annual Ion Channel Retreat, Vancouver, Canada, June 27-29, 2011.

Nine years ago Aurora Biomed Inc. (Vancouver, Canada) committed to gathering the brightest minds and the most innovative research companies at one conference. The Ion Channel Retreat provides a podium for scientific discourse spanning a wide range of ion channel disciplines.

The eighth annual ion channel retreat, Vancouver, Canada, June 28-30, 2010.

Eight years ago Aurora Biomed Inc. (Vancouver, Canada) committed to gathering the brightest minds and the most innovative research companies at one conference. We sought to provide a podium for scientific discourse spanning a wide range of ion channel disciplines.

Validation of a Rb+ uptake assay for the mouse embryonic stem cell-derived cardiomyocytes Na+, K+ ATPase.

Human Na+, K+ ATPase, an ATP-driven ion transporter, is an emerging drug target for heart-related conditions. Three types of assays including purified enzyme, radiotracer flux, and cold Rb+ flux have been used to determine the activity of this transporter. As an alternative to primary cardiomyocytes, mouse embryonic stem cells-derived cardiomyocytes with functional expression of essential cardiac ion channels were used in the present studies.

A cell-based Rb(+)-flux assay of the Kv1.3 potassium channel.

The Kv1.3 channels expressed by human T lymphocytes are emerging as important therapeutic targets. Peptides like agitoxin and margatoxin in scorpion venom and some non-peptide small molecules are known to inhibit this channel.

Development and validation of HTS flux assay for endogenously expressed chloride channels in a CHO-K1 cell line.

An atomic absorption spectroscopy-based detection system was employed to develop a new non-radioactive flux assay for chloride (Cl-) channels in a high throughput format. Cl- flux is assayed by measuring the extent to which Cl- precipitates an excess amount of silver ions (Ag+). A linear correlation was observed between theoretical and determined Cl- concentration with an r2 value of 0.

Evaluation of functional and binding assays in cells expressing either recombinant or endogenous hERG channel.

Introduction: The hERG (human ether-a-go-go related gene) potassium channel is required for normal cardiac repolarization, is susceptible to inhibition by a wide variety of compounds, and its blockage can lead to cardiac QT interval prolongation and life threatening arrhythmias. The present report examines the ability of hERG binding and functional assays to identify compounds with potential cardiovascular liabilities at the earliest stages of drug discovery.

Methods: Competitive binding assays were developed using (3)H-dofetilide and membranes from HEK293EBNA cells stably expressing recombinant hERG (HEK293-hERG) and IMR-32 cells expressing hERG endogenously.

Development of an HTS assay for Na+, K+-ATPase using nonradioactive rubidium ion uptake.

A high-throughput screening (HTS) assay was developed for the Na(+),K(+)-ATPase channel in order to study rubidium uptake as a measure of the functional activity and modulation of this exchanger. The assay uses elemental rubidium as a tracer for K(+) ions. Three cell lines were used to study the exchanger, and the assay was performed in a 96-well microtiter plate format.

Flux assays in high throughput screening of ion channels in drug discovery.

Ion channels have been identified as therapeutic targets in various disorders, such as cardiovascular disease, neurological disease, and cystic fibrosis. Flux assays to detect functional ionic flux through ion channels are becoming increasingly popular as tools for screening compounds. In an optimized flux assay, modulation of ion channel activity may produce readily detectable changes in radiolabeled or nonradiolabeled ionic flux.