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ORIGINAL RESEARCH article

Front. Cell. Infect. Microbiol.

Sec. Clinical and Diagnostic Microbiology and Immunology

Volume 15 - 2025 | doi: 10.3389/fcimb.2025.1643991

Multicenter evaluation of fast multiplex PCR for detection of pathogens in lower respiratory tract infections

Provisionally accepted
Ling WangLing Wang1Jingyao CaiJingyao Cai1Lizhong DaiLizhong Dai2Weimin MiaoWeimin Miao2Ziyang LiZiyang Li1Wei CaoWei Cao1Sisong HuangSisong Huang2Mei SunMei Sun1Lehuan XiaLehuan Xia3Xixin JiangXixin Jiang4Pingbang WangPingbang Wang5Jianhua PanJianhua Pan6Qinghe YangQinghe Yang7Anqun YangAnqun Yang7Min HuMin Hu1*
  • 1Second Xiangya Hospital, Central South University, Changsha, China
  • 2Sansure Biotech Inc., Changsha, China
  • 3Chenzhou Third People’s Hospital, Chenzhou, China
  • 4Yueyang People's Hospital, Yueyang, China
  • 5Liuyang People Hospital, Liuyang, China
  • 6Changsha Central Hospital, Changsha, China
  • 7The Second People's Hospital of Hunan Province Huaihua City, Huaihua, China

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Background: This study aimed to compare the diagnostic performance between bacterial culture and a rapid multiplex PCR assay (Respiratory Pathogens Multiplex Nucleic Acid Diagnostic Kit) for detecting six bacterial (Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus, Streptococcus pneumoniae, Haemophilus influenzae, Legionella pneumophila) and six viral targets (Influenza A/B, Respiratory syncytial virus, Adenoviruses, Human rhinovirus, Mycoplasma pneumoniae) in 728 bronchoalveolar lavage (BAL) specimens. Methods: This multicentric observational study was conducted at six comprehensive large hospitals in Hunan Province, China (May to October 2023) and assessed performance of mPCR kit by comparison with conventional culture method. Results: The mPCR kit detected ≥1 pathogen in 628 specimens (positivity rate: 86.3%), demonstrating positive percentage agreement (PPA) of 84.6% (95% CI: 76.6-92.6%) and negative percentage agreement (NPA) of 96.5% (95% CI: 96.0-97.1%) versus culture. Notably, semi-quantitative concordance was 79.3% (283/357) for culture-positive specimens. Multiple pathogens were detected by mPCR in 144 samples (19.8%). Lower Ct values (≤30) correlated strongly with culture positivity.

Keywords: multicenter evaluation, Lower respiratory tract infections, Pathogen Detection, multiplex PCR, Semi-quantitative result

Received: 09 Jun 2025; Accepted: 04 Sep 2025.

Copyright: © 2025 Wang, Cai, Dai, Miao, Li, Cao, Huang, Sun, Xia, Jiang, Wang, Pan, Yang, Yang and Hu. 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: Min Hu, Second Xiangya Hospital, Central South University, Changsha, 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.

Supplementary Material