Clinical validation of a novel multiplex real-time PCR for diagnosing community-acquired pneumonia in the emergency department: a promising diagnostic tool for Mycoplasma pneumoniae
- 1Department of Laboratory Medicine, The First Medical Center, Chinese PLA General Hospital
- 2Department of Emergency, The First Medical Center, Chinese PLA General Hospital, Medical School of Chinese PLA
Abstract
Community-acquired pneumonia (CAP) is a global public health burden with high morbidity, but timely diagnosis is hindered by limitations of conventional methods. We validated a novel multiplex real-time PCR (mRT-PCR) for CAP diagnosis in the emergency department, focusing on Mycoplasma pneumoniae ( M. pneumoniae ). A retrospective cohort of 371 CAP patients (2019–2021) from the Chinese PLA General Hospital was enrolled. The panel targeted 85 pathogens (40 bacteria, 18 viruses, 17 fungi, 10 other) plus 8 resistance genes, with a turnaround time (TAT) under 4 h and a cost of approximately US$100 per test. mRT-PCR showed performance comparable to mNGS for common pathogens (concordance >69% for bacteria/fungi) and high concordance with the final clinical diagnosis. Notably, it detected M. pneumoniae in 36.7% (136/371) of cases, outperforming conventional methods by identifying 204 exclusive positives (55.0% of cohort), with M. pneumoniae (40%) and viruses (30%) predominating. Polymicrobial infections occurred in 36.1% of cases; M. pneumoniae co-infections (38 cases) showed elevated neutrophils and reduced lymphocytes vs. single infections. An independent reference standard analysis excluding both index tests showed that 24.1% (210/873) of calls were independently corroborated - high for influenza, SARS-CoV-2 and M. pneumoniae , lower for S. pneumoniae , consistent with the limited sensitivity of sputum culture. Unlike conventional syndromic panels such as FilmArray, which profile fewer targets without concurrent resistance-gene detection, this mRT-PCR combines broad coverage with rapid TAT and low cost, making it promising for timely CAP diagnosis, particularly in resource-limited settings and for targeted therapy.
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BibTeX
@article{yang2026clinical,
title = {{Clinical validation of a novel multiplex real-time PCR for diagnosing community-acquired pneumonia in the emergency department: a promising diagnostic tool for Mycoplasma pneumoniae}},
author = {Liuyang Yang and Chaomin Guo and Qiang Zhao and Jiyong Yang and Heng Zhang and Yongzhi Zhai and Yanning Ma},
journal = {Frontiers in Microbiology},
year = {2026},
month = oct,
doi = {10.3389/fmicb.2026.1979614},
url = {https://www.frontiersin.org/articles/10.3389/fmicb.2026.1979614/full},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Clinical validation of a novel multiplex real-time PCR for diagnosing community-acquired pneumonia in the emergency department: a promising diagnostic tool for Mycoplasma pneumoniae
AU - Liuyang Yang
AU - Chaomin Guo
AU - Qiang Zhao
AU - Jiyong Yang
AU - Heng Zhang
AU - Yongzhi Zhai
AU - Yanning Ma
PY - 2026
DA - 2026/10/07
JO - Frontiers in Microbiology
DO - 10.3389/fmicb.2026.1979614
UR - https://www.frontiersin.org/articles/10.3389/fmicb.2026.1979614/full
AB - Community-acquired pneumonia (CAP) is a global public health burden with high morbidity, but timely diagnosis is hindered by limitations of conventional methods. We validated a novel multiplex real-time PCR (mRT-PCR) for CAP diagnosis in the emergency department, focusing on Mycoplasma pneumoniae ( M. pneumoniae ). A retrospective cohort of 371 CAP patients (2019–2021) from the Chinese PLA General Hospital was enrolled. The panel targeted 85 pathogens (40 bacteria, 18 viruses, 17 fungi, 10 other) plus 8 resistance genes, with a turnaround time (TAT) under 4 h and a cost of approximately US$100 per test. mRT-PCR showed performance comparable to mNGS for common pathogens (concordance >69% for bacteria/fungi) and high concordance with the final clinical diagnosis. Notably, it detected M. pneumoniae in 36.7% (136/371) of cases, outperforming conventional methods by identifying 204 exclusive positives (55.0% of cohort), with M. pneumoniae (40%) and viruses (30%) predominating. Polymicrobial infections occurred in 36.1% of cases; M. pneumoniae co-infections (38 cases) showed elevated neutrophils and reduced lymphocytes vs. single infections. An independent reference standard analysis excluding both index tests showed that 24.1% (210/873) of calls were independently corroborated - high for influenza, SARS-CoV-2 and M. pneumoniae , lower for S. pneumoniae , consistent with the limited sensitivity of sputum culture. Unlike conventional syndromic panels such as FilmArray, which profile fewer targets without concurrent resistance-gene detection, this mRT-PCR combines broad coverage with rapid TAT and low cost, making it promising for timely CAP diagnosis, particularly in resource-limited settings and for targeted therapy.
SN - 1664-302X
ER -