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Background <p>Difficulties in microbiologically confirming childhood tuberculosis (TB) can result in delayed treatment and increased disease severity.</p> Methods <p>In this study, we for the first time used whole genome next-generation sequencing (NGS) to detect cell-free DNA (cfDNA) from <i>Mycobacterium tuberculosis</i> (MTB) in plasma from children.</p> Results <p>We enrolled 94 children with active TB and 32 children with other respiratory infections. Combining NGS with probe capture enrichment (targeted cfNGS) showed higher coverage and detecting capability than did NGS alone. The targeted cfNGS showed slightly lower sensitivity (31.9% vs. 44.7%, <i>P</i> = 0.072) and specificity (96.9% vs. 100.0%, <i>P</i> = 0.236) to those of sputum tested using Xpert. Agreement between cfNGS-plasma and Xpert-sputum was weak (κ = 0.217). Concordant results were obtained for only 85 children (67.5%; 16 cases positive by both tests and 69 cases negative by both tests). A total of 40 children with MTB culture negative results were tested to have positive cfNGS-plasma or Xpert-sputum outcomes, yielding a significantly increased percentage of children with bacteriological evidence (20.2% [19/94] for MTB culture-positive only vs. 62.8% [59/94] for cfNGS-plasma, Xpert-sputum or culture positive).</p> Conclusions <p>These data suggest that cfNGS performed well for diagnosing TB using plasma from children. cfNGS may be a new method for diagnosing patients with paucibacillary TB.</p>

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Cell-free DNA next-generation sequencing for Mycobacterium tuberculosis obtained from plasma of children with active tuberculosis

  • 期刊论文

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  • 发表日期：

    2025年3月3日

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  • Shuting Quan,
  • Xue Tian,
  • Yuting Sun,
  • Hui Qi,
  • Weiwei Jiao,
  • Baixu Sun,
  • Fang Xu,
  • Min Fang,
  • Xuemei Yang,
  • Xi Zeng,
  • Kun Duan,
  • Jichao Wang,
  • Xue Fu,
  • Li Duan,
  • Lin Sun,
  • Adong Shen

摘要

Background

Difficulties in microbiologically confirming childhood tuberculosis (TB) can result in delayed treatment and increased disease severity.

Methods

In this study, we for the first time used whole genome next-generation sequencing (NGS) to detect cell-free DNA (cfDNA) from Mycobacterium tuberculosis (MTB) in plasma from children.

Results

We enrolled 94 children with active TB and 32 children with other respiratory infections. Combining NGS with probe capture enrichment (targeted cfNGS) showed higher coverage and detecting capability than did NGS alone. The targeted cfNGS showed slightly lower sensitivity (31.9% vs. 44.7%, P = 0.072) and specificity (96.9% vs. 100.0%, P = 0.236) to those of sputum tested using Xpert. Agreement between cfNGS-plasma and Xpert-sputum was weak (κ = 0.217). Concordant results were obtained for only 85 children (67.5%; 16 cases positive by both tests and 69 cases negative by both tests). A total of 40 children with MTB culture negative results were tested to have positive cfNGS-plasma or Xpert-sputum outcomes, yielding a significantly increased percentage of children with bacteriological evidence (20.2% [19/94] for MTB culture-positive only vs. 62.8% [59/94] for cfNGS-plasma, Xpert-sputum or culture positive).

Conclusions

These data suggest that cfNGS performed well for diagnosing TB using plasma from children. cfNGS may be a new method for diagnosing patients with paucibacillary TB.