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Evaluation of a fully automatic nucleic acid amplification system for detecting four respiratory pathogens

Pediatric Research (2025)Cite this article

Abstract

Background

Respiratory syncytial virus (RSV), adenovirus (ADV), human parainfluenza virus (hPIV), and Mycoplasma pneumoniae (MP) are prevalent pathogens causing acute respiratory infections (ARIs) in children. Prompt and precise detection of these pathogens is essential for early differentiation. This study sought to assess the diagnostic efficacy of a fully automatic real-time fluorescence PCR assay utilizing microfluidic technology (PCR-MT) for the rapid detection of RSV, hPIV, ADV, and MP in children in a hospital setting in Zhejiang, China.

Methods

The study was conducted on 420 children with ARIs from March to December 2022 at our hospital. Throat swab samples were collected and detected for RSV, hPIV, ADV, and MP using both PCR-capillary electrophoresis fragment analysis (PCR-CEFA) and PCR-MT. The results obtained from the PCR-MT method were compared with those from PCR-CEFA.

Results

With PCR-CEFA as the gold standard, the sensitivity and specificity of the PCR-MT method were as follows: 94.4% and 100.0% for RSV, 96.0% and 99.1% for hPIV, 100.0% and 98.6% for ADV, and 93.5% and 98.8% for MP, respectively.

Conclusion

The PCR-MT method demonstrates substantial potential for clinical application in the early diagnosis of RSV, hPIV, ADV, and MP in an outpatient setting, offering robust sensitivity and specificity.

Impact

  • Rapid, accurate, and convenient multiple pathogen detection technologies represent a significant area of research in the medical field. The method evaluated in this study enables simultaneous detection of four pathogens on a single chip, covering various subtypes, with results available within half an hour. Although some multi-pathogen detection chips are already commercially available, they may still have limitations such as sensitivity, specificity, and cost. Ongoing technological advancements could make pathogen detection more efficient, accurate, and economical. Continued attention to the development, validation, and optimization of these technologies in clinical practice will better serve patients.

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Data availability

The data presented in this study are available upon reasonable request from the corresponding author.

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Acknowledgements

We are grateful to the Department of Clinical Laboratory at The Children’s Hospital of Zhejiang University School of Medicine for supplying the essential laboratory equipment and location required for the collection of experimental data.

Funding

The study was supported by the Natural Science Foundation of Zhejiang Province, China under Grant No.ZCLTGC24H2001 and Medical Health Science and Technology Project of Zhejiang Provincial Health Commission under Grant No.WKJ-ZJ-2534.

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Authors and Affiliations

  1. Department of Clinical Laboratory, The Children’s Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China

    Ying-ying Chen, Wen-qing xiang, Ya-jun Guo, Zheng Shen & Wei Li

Authors
  1. Ying-ying Chen
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  2. Wen-qing xiang
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  3. Ya-jun Guo
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  4. Zheng Shen
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Contributions

Ying-ying Chen, Formal analysis, Investigation, Writing – original draft, Writing – review and editing. Wen-qing Xiang, Data curation, Formal analysis, Software, Validation, Writing – review and editing. Ya-jun Guo, Investigation, Writing – review and editing. Zheng Shen, Investigation, Writing – review and editing. Wei Li,Writing - review & editing, Supervision, Investigation, Formal analysis, Conceptualization. All authors have read and approved the final version of the manuscript.

Corresponding author

Correspondence to Wei Li.

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Competing interests

The authors declare no competing interests.

Ethics approval and consent to participate

The study’s design and methodology received approval from the Ethics Committee of the Children’s Hospital at Zhejiang University School of Medicine (Approval No. 2023-IRB-0112-Y-01). Given that the samples used in this research were residual from routine clinical tests, obtaining informed consent was exempted as per the committee’s guidelines. This research adheres to the principles of the Declaration of Helsinki.

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Chen, Yy., xiang, Wq., Guo, Yj. et al. Evaluation of a fully automatic nucleic acid amplification system for detecting four respiratory pathogens. Pediatr Res (2025). https://doi.org/10.1038/s41390-025-04101-1

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