Detection of Mycobacterium tuberculosis from tongue swabs using sonication and sequence-specific hybridization capture.

Autor: Yan AJ; Department of Bioengineering, University of Washington, Seattle, Washington, United States of America., Olson AM; Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, Washington, United States of America., Weigel KM; Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, Washington, United States of America., Luabeya AK; South African Tuberculosis Vaccine Initiative, Institute of Infectious Disease & Molecular Medicine and Department of Pathology, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa., Heiniger E; Department of Bioengineering, University of Washington, Seattle, Washington, United States of America., Hatherill M; South African Tuberculosis Vaccine Initiative, Institute of Infectious Disease & Molecular Medicine and Department of Pathology, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa., Cangelosi GA; Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, Washington, United States of America., Yager P; Department of Bioengineering, University of Washington, Seattle, Washington, United States of America.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2024 Aug 15; Vol. 19 (8), pp. e0308235. Date of Electronic Publication: 2024 Aug 15 (Print Publication: 2024).
DOI: 10.1371/journal.pone.0308235
Abstrakt: Tongue swabs hold promise as a non-invasive sample for diagnosing tuberculosis (TB). However, their utility as replacements for sputum has been limited by their varied diagnostic performance in PCR assays compared to sputum. The use of silica-based DNA extraction methods may limit sensitivity due to incomplete lysis of Mycobacterium tuberculosis (MTB) cells and co-extraction of non-target nucleic acid, which may inhibit PCR. Specificity may also be compromised because these methods are labor-intensive and prone to cross-contamination. To address these limitations, we developed a sample preparation method that combines sonication for MTB lysis and a sequence-specific MTB DNA capture method using hybridization probes immobilized on magnetic beads. In spiked tongue swabs, our hybridization capture method demonstrated a 100-fold increase in MTB DNA yield over silica-based Qiagen DNA extraction and ethanol precipitation. In a study conducted on clinical samples from South Africa, our protocol had 74% (70/94) sensitivity and 98% (41/42) specificity for detecting active pulmonary TB with sputum Xpert MTB/RIF Ultra as the reference standard. While hybridization capture did not show improved sensitivity over Qiagen DNA extraction and ethanol precipitation, it demonstrated better specificity than previously reported methods and was easier to perform. With integration into point-of-care platforms, these strategies have the potential to help enable rapid non-sputum-based TB diagnosis across key underserved patient populations.
Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: Paul Yager has a nonpaying appointment as CSO of the UbiDX corporation. There are no patents, products in development, or marketed products associated with this research to declare. Swabs were kindly donated by Copan. This does not alter our adherence to PLOS ONE policies on sharing data and materials.
(Copyright: © 2024 Yan et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
Databáze: MEDLINE
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