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Purpose <p><i>Mycoplasma pneumoniae</i>, a globally prevalent cause of community-acquired pneumonia in children and young adults, is typically diagnosed using molecular methods, including DNA- or RNA-based nucleic acid amplification tests (NAATs). While RNA-based NAAT is considered to reflect the viability and replication status of <i>M. pneumoniae</i>, a direct comparison of the clinical performance between DNA- and RNA-based NAATs has been lacking. This study aimed to compare these NAATs in clinical pediatric <i>M. pneumoniae</i> samples and elucidate the reasons for their differential detection outcomes.</p> Methods <p>This study analyzed clinical <i>M. pneumoniae</i> samples from pediatric patients across different sample types, seasons, age, and sex subgroups between January and December 2018. The underlying reasons for their differential detection outcomes were further elucidated through in vitro culture and cell infection models.</p> Results <p>A total of 3180 clinical samples were analyzed. The performance of DNA- and RNA-based NAATs showed specific disparities, particularly associated with sample type and patient age, but not with sampling season or patient sex. In vitro experiments revealed that <i>M. pneumoniae</i>-RNA (MP-RNA) has higher synthesis and degradation rates, whereas <i>M. pneumoniae</i>-DNA (MP-DNA) accumulates and is sustained longer in the growth dynamics of the pathogen.</p> Conclusion <p>The differential nucleic acid kinetics of <i>M. pneumoniae</i> across respiratory microenvironments likely explain the observed variations in clinical detection. These findings may enable evidence-based selection of DNA- or RNA-based NAATs according to patient age and sample type, thereby improving the reliable detection of <i>M. pneumoniae</i> in children.</p>

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Comparison between DNA- and RNA-based nucleic acid amplification tests for detecting Mycoplasma pneumoniae in pediatric specimens

  • 期刊论文

  • 发表日期：

    2026年2月28日

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  • Boyi Jiang,
  • Hanqing Zhao,
  • Chao Yan,
  • Mingxuan Wang,
  • Zhen Wang,
  • Yanling Feng,
  • Shijie Wang,
  • Jing Yuan,
  • Yuehua Ke

摘要

Purpose

Mycoplasma pneumoniae, a globally prevalent cause of community-acquired pneumonia in children and young adults, is typically diagnosed using molecular methods, including DNA- or RNA-based nucleic acid amplification tests (NAATs). While RNA-based NAAT is considered to reflect the viability and replication status of M. pneumoniae, a direct comparison of the clinical performance between DNA- and RNA-based NAATs has been lacking. This study aimed to compare these NAATs in clinical pediatric M. pneumoniae samples and elucidate the reasons for their differential detection outcomes.

Methods

This study analyzed clinical M. pneumoniae samples from pediatric patients across different sample types, seasons, age, and sex subgroups between January and December 2018. The underlying reasons for their differential detection outcomes were further elucidated through in vitro culture and cell infection models.

Results

A total of 3180 clinical samples were analyzed. The performance of DNA- and RNA-based NAATs showed specific disparities, particularly associated with sample type and patient age, but not with sampling season or patient sex. In vitro experiments revealed that M. pneumoniae-RNA (MP-RNA) has higher synthesis and degradation rates, whereas M. pneumoniae-DNA (MP-DNA) accumulates and is sustained longer in the growth dynamics of the pathogen.

Conclusion

The differential nucleic acid kinetics of M. pneumoniae across respiratory microenvironments likely explain the observed variations in clinical detection. These findings may enable evidence-based selection of DNA- or RNA-based NAATs according to patient age and sample type, thereby improving the reliable detection of M. pneumoniae in children.