IGI-LuNER: single-well multiplexed RT-qPCR test for SARS-CoV-2

Area of Science:

• Molecular Biology
• Virology
• Diagnostic Assays

Background:

• Current SARS-CoV-2 RT-qPCR diagnostics often require multiple reactions, increasing time and cost.
• Reliance on single viral targets in some tests elevates the risk of false-negative results.
• A single-tube assay combining multiple SARS-CoV-2 targets and a sample control is lacking.

Approach:

• Developed the IGI-LuNER RT-qPCR assay for simultaneous detection of SARS-CoV-2 N gene, E gene, and human RNase P (NER).
• Utilized the Luna Probe Universal One-Step RT-qPCR master mix with established primers and probes.
• Optimized for performance in 384-well plates.

Key Points:

• The IGI-LuNER assay integrates detection of two key SARS-CoV-2 genes (N and E) and a human sample control (RNase P) into a single reaction.
• This multiplexed approach is cost-effective and maintains high detection sensitivity suitable for clinical reporting.
• The assay is compatible with high-throughput platforms like 384-well plates.

Conclusions:

• The IGI-LuNER assay offers a streamlined, efficient, and accurate method for SARS-CoV-2 detection.
• This single-reaction assay reduces diagnostic time, cost, and the likelihood of false negatives.
• It facilitates future sample pooling strategies, significantly enhancing SARS-CoV-2 testing throughput.