Characterization of Drug-Resistant Lipid-Dependent Differentially Detectable <em>Mycobacterium tuberculosis</em>
An estimated 15–20% of patients who are treated for pulmonary tuberculosis (TB) are culture-negative at the time of diagnosis. Recent work has focused on the existence of differentially detectable <i>Mycobacterium tuberculosis</i> (<i>Mtb</i>) bacilli that do not grow under r...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-07-01
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| Series: | Journal of Clinical Medicine |
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| Online Access: | https://www.mdpi.com/2077-0383/10/15/3249 |
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| author | Annelies W. Mesman Seung-Hun Baek Chuan-Chin Huang Young-Mi Kim Sang-Nae Cho Thomas R. Ioerger Nadia N. Barreda Roger Calderon Christopher M. Sassetti Megan B. Murray |
| author_facet | Annelies W. Mesman Seung-Hun Baek Chuan-Chin Huang Young-Mi Kim Sang-Nae Cho Thomas R. Ioerger Nadia N. Barreda Roger Calderon Christopher M. Sassetti Megan B. Murray |
| author_sort | Annelies W. Mesman |
| collection | DOAJ |
| description | An estimated 15–20% of patients who are treated for pulmonary tuberculosis (TB) are culture-negative at the time of diagnosis. Recent work has focused on the existence of differentially detectable <i>Mycobacterium tuberculosis</i> (<i>Mtb</i>) bacilli that do not grow under routine solid culture conditions without the addition of supplementary stimuli. We identified a cohort of TB patients in Lima, Peru, in whom acid-fast bacilli could be detected by sputum smear microscopy, but from whom <i>Mtb</i> could not be grown in standard solid culture media. When we attempted to re-grow <i>Mtb</i> from the frozen sputum samples of these patients, we found that 10 out of 15 could be grown in a glycerol-poor/lipid-rich medium. These fell into the following two groups: a subset that could be regrown in glycerol after “lipid-resuscitation”, and a group that displayed a heritable glycerol-sensitive phenotype that were unable to grow in the presence of this carbon source. Notably, all of the glycerol-sensitive strains were found to be multidrug resistant. Although whole-genome sequencing of the lipid-resuscitated strains identified 20 unique mutations compared to closely related strains, no single genetic lesion could be associated with this phenotype. In summary, we found that lipid-based media effectively fostered the growth of <i>Mtb</i> from a series of sputum smear-positive samples that were not culturable in glycerol-based Lowenstein–Jensen or 7H9 media, which is consistent with <i>Mtb</i>’s known preference for non-glycolytic sources during infection. Analysis of the recovered strains demonstrated that both genetic and non-genetic mechanisms contribute to the observed differential capturability, and suggested that this phenotype may be associated with drug resistance. |
| first_indexed | 2024-03-10T09:14:27Z |
| format | Article |
| id | doaj.art-d5438668859341c587b34becdf0fb1a0 |
| institution | Directory Open Access Journal |
| issn | 2077-0383 |
| language | English |
| last_indexed | 2024-03-10T09:14:27Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Clinical Medicine |
| spelling | doaj.art-d5438668859341c587b34becdf0fb1a02023-11-22T05:48:03ZengMDPI AGJournal of Clinical Medicine2077-03832021-07-011015324910.3390/jcm10153249Characterization of Drug-Resistant Lipid-Dependent Differentially Detectable <em>Mycobacterium tuberculosis</em>Annelies W. Mesman0Seung-Hun Baek1Chuan-Chin Huang2Young-Mi Kim3Sang-Nae Cho4Thomas R. Ioerger5Nadia N. Barreda6Roger Calderon7Christopher M. Sassetti8Megan B. Murray9Gemeentelijke Gezondheidsdiensten en Geneeskundige Hulpverleningsorganisaties in de Regio Nederland, 3524 SJ Utrecht, The NetherlandsDepartment of Microbiology and Institute for Immunology and Immunological Diseases, College of Medicine, Yonsei University, Seoul 16995, KoreaDivision of Global Health Equity, Brigham and Women’s Hospital, Boston, MA 02115, USADepartment of Microbiology and Institute for Immunology and Immunological Diseases, College of Medicine, Yonsei University, Seoul 16995, KoreaDepartment of Microbiology and Institute for Immunology and Immunological Diseases, College of Medicine, Yonsei University, Seoul 16995, KoreaDepartment of Computer Science and Engineering, Texas A&M University, College Station, TX 77843, USASocios En Salud Sucursal Peru, Lima 15001, PeruSocios En Salud Sucursal Peru, Lima 15001, PeruDepartment of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA 01655, USADepartment of Global Health and Social Medicine, Harvard Medical School, Boston, MA 02115, USAAn estimated 15–20% of patients who are treated for pulmonary tuberculosis (TB) are culture-negative at the time of diagnosis. Recent work has focused on the existence of differentially detectable <i>Mycobacterium tuberculosis</i> (<i>Mtb</i>) bacilli that do not grow under routine solid culture conditions without the addition of supplementary stimuli. We identified a cohort of TB patients in Lima, Peru, in whom acid-fast bacilli could be detected by sputum smear microscopy, but from whom <i>Mtb</i> could not be grown in standard solid culture media. When we attempted to re-grow <i>Mtb</i> from the frozen sputum samples of these patients, we found that 10 out of 15 could be grown in a glycerol-poor/lipid-rich medium. These fell into the following two groups: a subset that could be regrown in glycerol after “lipid-resuscitation”, and a group that displayed a heritable glycerol-sensitive phenotype that were unable to grow in the presence of this carbon source. Notably, all of the glycerol-sensitive strains were found to be multidrug resistant. Although whole-genome sequencing of the lipid-resuscitated strains identified 20 unique mutations compared to closely related strains, no single genetic lesion could be associated with this phenotype. In summary, we found that lipid-based media effectively fostered the growth of <i>Mtb</i> from a series of sputum smear-positive samples that were not culturable in glycerol-based Lowenstein–Jensen or 7H9 media, which is consistent with <i>Mtb</i>’s known preference for non-glycolytic sources during infection. Analysis of the recovered strains demonstrated that both genetic and non-genetic mechanisms contribute to the observed differential capturability, and suggested that this phenotype may be associated with drug resistance.https://www.mdpi.com/2077-0383/10/15/3249diagnosistuberculosisTBculturedrug-resistant |
| spellingShingle | Annelies W. Mesman Seung-Hun Baek Chuan-Chin Huang Young-Mi Kim Sang-Nae Cho Thomas R. Ioerger Nadia N. Barreda Roger Calderon Christopher M. Sassetti Megan B. Murray Characterization of Drug-Resistant Lipid-Dependent Differentially Detectable <em>Mycobacterium tuberculosis</em> Journal of Clinical Medicine diagnosis tuberculosis TB culture drug-resistant |
| title | Characterization of Drug-Resistant Lipid-Dependent Differentially Detectable <em>Mycobacterium tuberculosis</em> |
| title_full | Characterization of Drug-Resistant Lipid-Dependent Differentially Detectable <em>Mycobacterium tuberculosis</em> |
| title_fullStr | Characterization of Drug-Resistant Lipid-Dependent Differentially Detectable <em>Mycobacterium tuberculosis</em> |
| title_full_unstemmed | Characterization of Drug-Resistant Lipid-Dependent Differentially Detectable <em>Mycobacterium tuberculosis</em> |
| title_short | Characterization of Drug-Resistant Lipid-Dependent Differentially Detectable <em>Mycobacterium tuberculosis</em> |
| title_sort | characterization of drug resistant lipid dependent differentially detectable em mycobacterium tuberculosis em |
| topic | diagnosis tuberculosis TB culture drug-resistant |
| url | https://www.mdpi.com/2077-0383/10/15/3249 |
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