Multiplex Quantitative Polymerase Chain Reaction Test to Identify SARS-CoV-2 Variants.

Area of Science:

• Molecular Biology
• Virology
• Genetics

Background:

• Quantitative polymerase chain reaction (qPCR) is a standard technique for real-time gene expression analysis.
• Multiplexing in qPCR utilizes multiple fluorescent probes for simultaneous detection of different targets.
• The emergence of SARS-CoV-2 variants necessitates rapid and accurate detection methods.

Purpose of the Study:

• To describe a novel method for simultaneous real-time quantitation of specific SARS-CoV-2 variants.
• To evaluate the utility of an Inhibitor-Tolerant RT-qPCR kit for variant detection.

Main Methods:

• Utilized quantitative polymerase chain reaction (qPCR) with fluorescent reporter probes.
• Employed an Inhibitor-Tolerant RT-qPCR kit for enhanced reaction efficiency.
• Developed a multiplexing approach for simultaneous detection of multiple targets.

Main Results:

• The described method enables simultaneous real-time quantitation of SARS-CoV-2 variants.
• The Inhibitor-Tolerant RT-qPCR kit demonstrated efficacy in detecting UK, South Africa, and Brazil variants.
• Multiplexing allowed for unique fluorescent signatures for each detected amplicon.

Conclusions:

• The developed qPCR method offers a robust approach for simultaneous variant surveillance.
• The Inhibitor-Tolerant RT-qPCR kit is suitable for real-time detection of multiple SARS-CoV-2 strains.
• This technique can aid in monitoring the spread of specific viral lineages.