Performance Evaluation of Nucleic Acid Detection Reagents and Verification of Various Factors Affecting the Efficiency of Nucleic.

Background: Since the COVID-19 outbreak in 2019, nucleic acid testing has been central to epidemic control and clinical decision-making. However, variability in assay kits and extraction steps can affect sensitivity, specificity, and reproducibility. Systematic evaluation of detection reagents and extraction efficiency is essential to ensure reliable clinical results. Objective: This study aimed to evaluate whether three domestic novel coronavirus nucleic acid detection kits based on fluorescence PCR meet clinical requirements for COVID-19 testing. Materials and Methods: The performance of the kits was assessed through coincidence rate, precision, detection limit, specificity, and interference resistance using standard positive, negative, and interference reference samples. Amplification efficiency of three detection reagents was compared with eight positive clinical samples. Additionally, amplification performance was examined under oscillation and non-oscillation conditions before nucleic acid extraction, and with varying input volumes. Results: