Development, Analytical, and Clinical Evaluation of Rapid Immunochromatographic Antigen Test for SARS-CoV-2 Variants Detection

Abstract

The antigen rapid diagnostic test (Ag-RDT) is a useful diagnostic tool for the detection and management of COVID-19 spread. Global SARS-CoV-2 variant outbreaks have highlighted the need for a test capable of detecting SARS-CoV-2 variants with high sensitivity and a low limit of detection. This study aimed to develop and evaluate, both analytically and clinically, an antigen rapid diagnostic test (the KestrelTM COVID-19 Ag Rapid Test) for professional use. A lateral flow immunoassay-based diagnostic test kit was developed, and various aspects of its analytical performance were evaluated. This test kit was clinically evaluated by two independent laboratories and showed closely related results of 96.49% and 98.33% of sensitivity, 100% and 100% of specificity, and 99.01% and 99.44% of accuracy, respectively. A limit of detection was observed at values as low as 0.156 ng/mL for recombinant SARS-CoV-2 nucleocapsid protein. Moreover, the test kit successfully detected the recombinant SARS-CoV-2 nucleocapsid protein (NP) of wild-type, Alpha-, Beta-, Gamma-, Delta-, Epsilon-, Kappa-, and Omicron-variants as positive results. Therefore, the KestrelTM COVID-19 Ag Rapid Test may have potential use for effective COVID-19 screening, surveillance, and infection control in a variety of global SARS-CoV-2 variant outbreaks.

Asia Pacific perspectives on the second year of the COVID‐19 pandemic: A follow‐up survey

“…Thirdly, diagnostic tests like antigen rapid tests (ART) 31 do have increased sensitivity of early diagnosis of asymptomatic or mildly symptomatic persons when community prevalence is high. They appear to be able to pick up variants like Omicron with high sensitivity, although results differ depending on different test kits 32 . This allows for early self‐detection, short test turnaround time and reduction in manpower and infrastructure needed to administer and run PCR tests.…”

Development of an Antigen Capture Lateral Flow Immunoassay for the Detection of Burkholderia pseudomallei

“…The gliding was carried out at a flow rate of 100 mm/second and with a volume of 1 µL/cm using a Kinbio reagent dispenser (Shanghai Kinbio Tech., Shanghai, China). After drying, the membranes were assembled with an Ahlstrom 8964 conjugate pad (Ahlstrom, Helsinki, Finland), a Millipore C048 sample pad (Merck KGaA, Darmstadt, Germany), and a Whatman 470 absorbance pad (Whatman, Buckinghamshire, UK), with each pad, overlapped by approximately 2 mm [16,24,25]. The assembled cards were then cut into 4 mm-width strips.…”

“…Bp2.1 mAb was conjugated to 40 nm colloidal gold nanoparticles (Kestrel BioSciences, Pathum Thani, Thailand) by passive absorption, blocked with PBS containing 10% bovine serum albumin, and concentrated to an OD 540 of 10 [24,25]. The gold-conjugated Bp2.1 (4 µL) was added to the conjugate pad prior to running the assay to serve as a detection antibody.…”

Catalytic Antibodies: Design, Expression, and Their Applications in Medicine

“…At the end of 2019, a new coronavirus spread over the globe, causing a new type of pneumonia disease (COVID-19) with high transmission and fatality [ 186 – 190 ]. A safe and effective vaccination is desperately needed, but development takes time.…”