Endogenous Biomarkers Analysis and False-Negative Results for SARSCov2 Using two Commercial RT-PCR Diagnostic Kits.

Area of Science:

• Virology
• Molecular Diagnostics
• Public Health

Background:

• Real-time PCR is standard for SARS-CoV-2 detection.
• False-negative results can occur despite internal controls.
• Investigating alternative internal controls is crucial for accuracy.

Purpose of the Study:

• To evaluate the impact of different internal control materials (DNA vs. RNA) on SARS-CoV-2 detection kit accuracy.
• To assess the reliability of DNA and RNA internal controls in COVID-19 diagnostics.
• To identify strategies for reducing false-negative results in SARS-CoV-2 testing.

Main Methods:

• 162 respiratory samples were tested using Pishtaz Teb (DNA control) and Geneova (RNA control) kits.
• Negative samples were retested with the Geneova kit for comparison.
• Positive and negative controls were used throughout the study.

Main Results:

• Only one false-negative result was identified after retesting with the RNA control.
• DNA and RNA controls showed agreement in only 44% of cases.
• Higher cycle threshold values for RNA controls indicated potential degradation.

Conclusions:

• Accurate SARS-CoV-2 detection relies on robust quality control.
• Selecting sensitive and specific diagnostic kits is vital to minimize false negatives.
• RNA internal controls can detect degradation, aiding in the identification of false negatives and improving disease management.