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Updated: May 7, 2025

Candidate Gene Testing in Clinical Cohort Studies with Multiplexed Genotyping and Mass Spectrometry
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Multigene Mycobacterium tuberculosis cell-free DNA assay.

S Ayalew1, T Wegayehu2, B Wondale2

  • 1Armauer Hansen Research Institute, Addis Ababa, Ethiopia;, Department of Biology, College of Natural and Computational Sciences, Arba Minch University, Arba Minch, Ethiopia.

The International Journal of Tuberculosis and Lung Disease : the Official Journal of the International Union Against Tuberculosis and Lung Disease
|January 4, 2025
PubMed
Summary
This summary is machine-generated.

A new plasma DNA test for tuberculosis (TB) shows promise. Targeting multiple genes in cell-free DNA (cfDNA) from TB patients improves diagnostic sensitivity, offering a potential alternative to sputum-based methods.

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Area of Science:

  • Medical Diagnostics
  • Molecular Biology
  • Infectious Diseases

Background:

  • Current tuberculosis diagnostics often rely on sputum, posing collection challenges.
  • Plasma Mycobacterium tuberculosis cell-free DNA (Mtb cfDNA) offers an alternative diagnostic target.

Purpose of the Study:

  • To evaluate a quantitative PCR (qPCR) assay using plasma Mtb cfDNA for pulmonary TB diagnosis.
  • To assess the sensitivity and specificity of targeting multiple genes (IS6110, cyp141, devR).

Main Methods:

  • Plasma samples from 106 pulmonary TB patients and 60 controls were analyzed.
  • A qPCR assay targeted IS6110, cyp141, and devR genes.
  • Sensitivity was compared against Xpert MTB/RIF, culture, and clinical diagnosis; specificity against controls.

Main Results:

  • The Mtb cfDNA assay achieved 71.7% sensitivity for all PTB cases, higher in bacteriologically confirmed (78.3%) than clinically diagnosed (28.6%) cases.
  • Combined specificity was 91.7%.
  • Targeting multiple genes, particularly IS6110 and cyp141 (70.8% sensitivity), improved overall detection rates compared to single-gene targets.

Conclusions:

  • Plasma Mtb cfDNA quantitative PCR is a potential adjunctive tool for TB diagnosis.
  • Multiplex gene targeting enhances the sensitivity of cfDNA-based TB diagnostics.
  • This approach may overcome limitations associated with sputum sample collection.