Novel Lateral Flow-Based Assay for Simple and Visual Detection of SARS-CoV-2 Mutations.

Area of Science:

• Molecular biology
• Virology
• Public Health

Background:

• Real-time identification of SARS-CoV-2 variants is crucial for effective virus control and public health responses.
• Reverse transcription quantitative polymerase chain reaction (RT-qPCR) is the standard for screening SARS-CoV-2 mutations but has limitations in multiplexing and susceptibility to new mutations.

Purpose of the Study:

• To present a proof-of-concept for a simple molecular assay for detecting key SARS-CoV-2 mutations.
• To develop and evaluate multiplex kits for variant surveillance.

Main Methods:

• Development of multiplex asymmetric one-step RT-PCR amplification targeting different regions of the SARS-CoV-2 S gene.
• Visual detection of mutations using a lateral flow DNA microarray.
• Clinical performance assessment using RNA extracts from 113 SARS-CoV-2-positive samples.

Main Results:

• The developed assay demonstrated specific and sensitive detection of key SARS-CoV-2 mutations.
• Performance was comparable to RT-qPCR for variant screening.
• The VAR-CoV assay successfully screened spike mutations associated with major variants of concern.

Conclusions:

• The assay is a promising tool for real-time SARS-CoV-2 variant surveillance.
• The assay can be rapidly adapted to detect emerging mutations.
• It offers a sensitive, specific, and visually detectable alternative for mutation screening.