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Faecal mtDNA heteroplasmy as a non-invasive biomarker in m.3243 A > G mitochondrial disease

Nature作者：Charlotte Warren2026年8月12日正文已收录本站
  • Article
  • Open access
  • Published:
  • Renae J. Stefanetti  ORCID: orcid.org/0000-0001-6402-64351,2 na1,
  • Isabel Barrow1,2,
  • David Houghton1,
  • Robert McFarland1,2,3,4,
  • Nell Richardson1,
  • Helen A. L. Tuppen1,
  • Yi Shiau Ng  ORCID: orcid.org/0000-0002-7591-20341,2,3,
  • Alasdair P. Blain  ORCID: orcid.org/0000-0002-3712-52831 &
  • …
  • Gráinne S. Gorman1 

Communications Medicine (2026) Cite this article

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Abstract

Background

Quantifying mitochondrial DNA (mtDNA) heteroplasmy is critical for diagnosis, prognosis, and monitoring of mitochondrial disease, particularly that caused by the m.3243 A > G pathogenic variant, where tissue-specific heteroplasmy contributes to disease expression. However, current methods often rely on invasive or clinic-based sampling, limiting accessibility and repeat testing. We validate a non-invasive, decentralised approach using faecal-derived mtDNA to quantify m.3243 A > G heteroplasmy.

Methods

Fifty participants with genetically confirmed m.3243 A > G mitochondrial disease were recruited remotely and sent a postal stool collection kit. DNA was extracted from returned stool samples and heteroplasmy at nucleotide position m.3243 A > G within MT-TL1 was quantified using pyrosequencing. Where available, stool mtDNA heteroplasmy was compared with previously collected measurements from muscle, blood, urine, and buccal swabs. Associations were assessed using Pearson correlation, agreement between measurements was evaluated using Bland-Altman analysis, and diagnostic performance for identifying high muscle heteroplasmy was assessed using receiver operating characteristic (ROC) analysis.

Results

Here we show that 47 of 50 participants return a sample and 44 of 47 yield an analysable result. Faecal m.3243 A > G and heteroplasmy correlates with muscle heteroplasmy (r = 0.638, R2 = 0.407, p = 0.0141). For identifying individuals with a high muscle heteroplasmy ( ≥ 60%), a faecal cut-off of ≥56.4% provides 83.3% sensitivity, 50% specificity, and a positive predictive value (PPV) of 90.9%, with high overall accuracy (area under the curve (AUC) 0.92).

Conclusions

We demonstrate that faecal mtDNA heteroplasmy is a promising biomarker and validate the feasibility of remote, postal home sample collection with central laboratory analysis aligned with decentralised precision medicine. This approach may reduce reliance on clinic-based sampling, broaden access to laboratory testing, and enable longitudinal monitoring and remote data collection in decentralised clinical trials.

Plain language summary

Mitochondrial disease caused by the m.3243 A > G genetic change is difficult to monitor because the level of change varies between tissues, and the most informative test often requires a muscle biopsy. We tested whether stool samples collected at home and returned to central laboratory could provide a simpler alternative. Fifty people in the United Kingdom were mailed a home collection kit. Returned samples were analysed to measure the proportion of the genetic change in human DNA present in stool, and we compared results with previous measurements from other tissues, including muscle. We found that stool testing provided usable results and that stool levels generally match with muscle levels. This approach could reduce invasive biopsies, improve access to testing, support repeat monitoring, and enable home-based clinical trials.

Subjects

Acknowledgements

We extend our sincere gratitude to all patients who participated in the study. We would like to extend our thanks to Laura Brown and Clare Massarella, who provided clinical trial management support, Doug Jerry and Qasim Haider for data management support and Dr James Stewart for his valuable contributions to the laboratory work and subsequent DNA analysis.

Funding

This study was funded by the Medical Research Council - Confidence in Concept award. This work was also supported by the Wellcome Trust [203105/A/16/Z] and infrastructural research support from the National Institute for Health and Care Research (NIHR) Biomedical Research Centre (BRC): Newcastle. The NIHR BRC: Newcastle is a partnership between Newcastle Hospitals NHS Foundation Trust, Newcastle University and Cumbria, Northumberland, Tyne and Wear NHS Foundation Trust funded by the NIHR. The views expressed are those of the author(s) and not necessarily those of the NIHR or the Department of Health and Social Care.

Author information

Author notes

  1. These authors contributed equally: Charlotte Warren, Renae J. Stefanetti.

Authors and Affiliations

  1. Translational and Clinical Research Institute. Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne, UK

    Charlotte Warren, Renae J. Stefanetti, Isabel Barrow, David Houghton, Robert McFarland, Nell Richardson, Helen A. L. Tuppen, Yi Shiau Ng, Alasdair P. Blain & Gráinne S. Gorman

  2. NIHR Biomedical Research Centre, Newcastle, UK

    Renae J. Stefanetti, Isabel Barrow, Robert McFarland & Yi Shiau Ng

  3. Directorate of Neurosciences, Royal Victoria Infirmary, Newcastle upon Tyne Hospitals NHS Foundation Trust, Newcastle upon Tyne, UK

    Robert McFarland & Yi Shiau Ng

  4. NHS Highly Specialised Service for Rare Mitochondrial Disorders of Adults and Children, Newcastle upon Tyne Hospitals NHS Foundation Trust, Newcastle upon Tyne, UK

    Robert McFarland

Authors

  1. Charlotte Warren
  2. Renae J. Stefanetti
  3. Isabel Barrow
  4. David Houghton
  5. Robert McFarland
  6. Nell Richardson
  7. Helen A. L. Tuppen
  8. Yi Shiau Ng
  9. Alasdair P. Blain
  10. Gráinne S. Gorman

Corresponding author

Correspondence to Renae J. Stefanetti.

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Competing interests

All authors declare no competing interests.

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Cite this article

Warren, C., Stefanetti, R.J., Barrow, I. et al. Faecal mtDNA heteroplasmy as a non-invasive biomarker in m.3243 A > G mitochondrial disease. Commun Med (2026). https://doi.org/10.1038/s43856-026-01847-5

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  • DOI: https://doi.org/10.1038/s43856-026-01847-5