ORIGINAL RESEARCH article
Front. Microbiol.
Sec. Virology
Volume 16 - 2025 | doi: 10.3389/fmicb.2025.1609142
Comprehensive Performance Evaluation of a High-Throughput Automated System for Pathogen Nucleic Acid Detection in Clinical Settings
Provisionally accepted
- 1Peking Union Medical College Graduate School, Beijing, China
- 2Peking Union Medical College Hospital (CAMS), Beijing, Beijing Municipality, China
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Objective: This study evaluates the clinical performance of a high-throughput automated molecular detection system and proposes a comprehensive and standardized performance validation framework to address gaps in existing methodologies and provide a robust reference for future evaluations.Performance was validated for EBV DNA, HCMV DNA, and RSV RNA using clinical samples at various concentrations, along with WHO and national reference standards. The validation included concordance rate, accuracy, linearity, precision, limit of detection, interference testing, cross-reactivity, and carryover contamination.The positive, negative, and overall concordance rates for EBV DNA, HCMV DNA, and RSV RNA were all 100%. Both intra-assay and inter-assay precision showed coefficients of variation (CV) below 5%. The linear correlation coefficient (|r|) for EBV DNA and HCMV DNA was ≥0.98, demonstrating excellent linearity. The limits of detection (LoD) were 10 IU/mL for EBV DNA and HCMV DNA, and 200 copies/mL for RSV RNA. Both interference and cross-reactivity assessments met the CLSI EP07 standards, and no carryover contamination was observed.The system demonstrated excellent performance in terms of concordance, accuracy, precision, linearity, interference testing, and cross-reactivity. It is highly suited for large-scale pathogen screening and routine nucleic acid testing in clinical laboratories, both for qualitative and quantitative analyses. Additionally, this study introduces a comprehensive and standardized performance validation framework that addresses critical gaps in existing methodologies, offering a robust foundation for the rigorous evaluation of diagnostic systems and serving as a valuable reference for future research.
Keywords: Clinical Detection of Pathogen Nucleic Acids, Automation, high-throughput, Performance verification, RT-PCR
Received: 10 Apr 2025; Accepted: 26 May 2025.
Copyright: © 2025 Lu, Zhang and Liu. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Rui Zhang, Peking Union Medical College Hospital (CAMS), Beijing, 100730, Beijing Municipality, China
Yali Liu, Peking Union Medical College Hospital (CAMS), Beijing, 100730, Beijing Municipality, China
Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.