Performance Characteristics of a High-Throughput Automated Transcription-Mediated Amplification Test for SARS-CoV-2...

Area of Science:

• Molecular diagnostics
• Virology
• Public health

Background:

• The COVID-19 pandemic necessitated rapid development of diagnostic tests for SARS-CoV-2.
• Laboratories faced challenges in scaling up diagnostic testing capacity.
• Automated molecular assays are crucial for high-throughput detection.

Purpose of the Study:

• To evaluate the analytical and clinical performance of a new automated nucleic acid amplification test system for SARS-CoV-2 detection.
• To assess the sensitivity, specificity, and reliability of the automated assay.

Main Methods:

• Utilized target capture, transcription-mediated amplification, and acridinium ester probe chemistry on the automated Panther system.
• Determined the limit of detection using inactivated virus and synthetic RNA targets.
• Assessed analytical sensitivity with commercial nucleic acid controls.
• Evaluated cross-reactivity and interference with other pathogens.
• Conducted clinical performance testing on nasopharyngeal swabs and compared results to RT-PCR.

Main Results:

• The limit of detection was as low as 0.004 TCID50/ml (virus) and 25 copies/ml (synthetic RNA).
• Analytical sensitivity confirmed 100% detection between 83-194 copies/ml.
• No cross-reactivity or interference observed with related coronaviruses and other pathogens.
• Clinical testing showed 100% positive, 98.7% negative, and 99.3% overall agreement with RT-PCR.

Conclusions:

• The automated system provides a sensitive and specific method for SARS-CoV-2 molecular diagnostics.
• This high-throughput assay is suitable for pandemic-scale automated testing.
• The validated performance supports its use in clinical diagnostic settings.