Differential Culturability of Mycobacterium tuberculosis in Culture-Negative Sputum of Patients With Pulmonary Tuberculosis and in a Simulated Model of Dormancy
Tuberculosis (TB) remains a leading killer among infectious diseases of humans worldwide. Delayed diagnosis is a crucial problem in global TB control programs. Bacteriological methods currently used to diagnose TB in endemic countries take up to 8 weeks, which poses a significant delay in starting a...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2019-10-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2019.02381/full |
| _version_ | 1818317884610314240 |
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| author | Azger Dusthackeer Magizhaveni Balasubramanian Govindarajan Shanmugam Shanmuga Priya Christy Rosaline Nirmal Rajadas Sam Ebenezer Angayarkanni Balasubramanian Rajesh Kumar Mondal Kannan Thiruvenkadam A. K. Hemanth Kumar Geetha Ramachandran Selvakumar Subbian |
| author_facet | Azger Dusthackeer Magizhaveni Balasubramanian Govindarajan Shanmugam Shanmuga Priya Christy Rosaline Nirmal Rajadas Sam Ebenezer Angayarkanni Balasubramanian Rajesh Kumar Mondal Kannan Thiruvenkadam A. K. Hemanth Kumar Geetha Ramachandran Selvakumar Subbian |
| author_sort | Azger Dusthackeer |
| collection | DOAJ |
| description | Tuberculosis (TB) remains a leading killer among infectious diseases of humans worldwide. Delayed diagnosis is a crucial problem in global TB control programs. Bacteriological methods currently used to diagnose TB in endemic countries take up to 8 weeks, which poses a significant delay in starting antibiotic therapy. The presence of a heterogeneous population of Mycobacterium tuberculosis, the causative agent of TB, is among the reasons for delayed diagnosis by bacteriological methods. Previously, it has been shown that mycobacterial resuscitation-promoting factors (RPFs), a family of proteins secreted by actively growing bacteria into the media, are capable of activating the growth of dormant bacteria, thus enhancing the detection of bacilli in the sputum of confirmed TB cases. However, the variability in bacterial resuscitation by RPF in the sputum of suspected pulmonary TB cases that showed differential smear and/or culture positivity during diagnosis has not been fully explored. Here, we report the presence of non-replicating bacteria in the sputum of suspected TB cases that show differential growth response to RPF treatment. Using crude and recombinant RPF treatment, we show improved sensitivity and reduced time to detect bacilli in the sputum samples of smear-positive/culture-negative or smear-negative/culture-negative cases. We also report the phenotypic heterogeneity in the RPF responsiveness among Mtb strains using an in vitro dormancy model. Our findings have implications for improving the bacteriological diagnostic modalities currently used to diagnose TB in endemic countries. |
| first_indexed | 2024-12-13T09:44:25Z |
| format | Article |
| id | doaj.art-b926df4005764f7a88291b9f76891c23 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-13T09:44:25Z |
| publishDate | 2019-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-b926df4005764f7a88291b9f76891c232022-12-21T23:52:06ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2019-10-011010.3389/fmicb.2019.02381492427Differential Culturability of Mycobacterium tuberculosis in Culture-Negative Sputum of Patients With Pulmonary Tuberculosis and in a Simulated Model of DormancyAzger Dusthackeer0Magizhaveni Balasubramanian1Govindarajan Shanmugam2Shanmuga Priya3Christy Rosaline Nirmal4Rajadas Sam Ebenezer5Angayarkanni Balasubramanian6Rajesh Kumar Mondal7Kannan Thiruvenkadam8A. K. Hemanth Kumar9Geetha Ramachandran10Selvakumar Subbian11National Institute for Research in Tuberculosis, Chennai, IndiaNational Institute for Research in Tuberculosis, Chennai, IndiaNational Institute for Research in Tuberculosis, Chennai, IndiaNational Institute of Epidemiology, Chennai, IndiaNational Institute for Research in Tuberculosis, Chennai, IndiaNational Institute for Research in Tuberculosis, Chennai, IndiaNational Institute for Research in Tuberculosis, Chennai, IndiaNational Institute for Research in Tuberculosis, Chennai, IndiaNational Institute for Research in Tuberculosis, Chennai, IndiaNational Institute for Research in Tuberculosis, Chennai, IndiaNational Institute for Research in Tuberculosis, Chennai, IndiaPublic Health Research Institute, New Jersey Medical School, Rutgers University, Newark, NJ, United StatesTuberculosis (TB) remains a leading killer among infectious diseases of humans worldwide. Delayed diagnosis is a crucial problem in global TB control programs. Bacteriological methods currently used to diagnose TB in endemic countries take up to 8 weeks, which poses a significant delay in starting antibiotic therapy. The presence of a heterogeneous population of Mycobacterium tuberculosis, the causative agent of TB, is among the reasons for delayed diagnosis by bacteriological methods. Previously, it has been shown that mycobacterial resuscitation-promoting factors (RPFs), a family of proteins secreted by actively growing bacteria into the media, are capable of activating the growth of dormant bacteria, thus enhancing the detection of bacilli in the sputum of confirmed TB cases. However, the variability in bacterial resuscitation by RPF in the sputum of suspected pulmonary TB cases that showed differential smear and/or culture positivity during diagnosis has not been fully explored. Here, we report the presence of non-replicating bacteria in the sputum of suspected TB cases that show differential growth response to RPF treatment. Using crude and recombinant RPF treatment, we show improved sensitivity and reduced time to detect bacilli in the sputum samples of smear-positive/culture-negative or smear-negative/culture-negative cases. We also report the phenotypic heterogeneity in the RPF responsiveness among Mtb strains using an in vitro dormancy model. Our findings have implications for improving the bacteriological diagnostic modalities currently used to diagnose TB in endemic countries.https://www.frontiersin.org/article/10.3389/fmicb.2019.02381/fulltuberculosisbacteriological diagnosisresuscitationMycobacteirum tuberculosisclinical isolate organismsputum samples |
| spellingShingle | Azger Dusthackeer Magizhaveni Balasubramanian Govindarajan Shanmugam Shanmuga Priya Christy Rosaline Nirmal Rajadas Sam Ebenezer Angayarkanni Balasubramanian Rajesh Kumar Mondal Kannan Thiruvenkadam A. K. Hemanth Kumar Geetha Ramachandran Selvakumar Subbian Differential Culturability of Mycobacterium tuberculosis in Culture-Negative Sputum of Patients With Pulmonary Tuberculosis and in a Simulated Model of Dormancy Frontiers in Microbiology tuberculosis bacteriological diagnosis resuscitation Mycobacteirum tuberculosis clinical isolate organism sputum samples |
| title | Differential Culturability of Mycobacterium tuberculosis in Culture-Negative Sputum of Patients With Pulmonary Tuberculosis and in a Simulated Model of Dormancy |
| title_full | Differential Culturability of Mycobacterium tuberculosis in Culture-Negative Sputum of Patients With Pulmonary Tuberculosis and in a Simulated Model of Dormancy |
| title_fullStr | Differential Culturability of Mycobacterium tuberculosis in Culture-Negative Sputum of Patients With Pulmonary Tuberculosis and in a Simulated Model of Dormancy |
| title_full_unstemmed | Differential Culturability of Mycobacterium tuberculosis in Culture-Negative Sputum of Patients With Pulmonary Tuberculosis and in a Simulated Model of Dormancy |
| title_short | Differential Culturability of Mycobacterium tuberculosis in Culture-Negative Sputum of Patients With Pulmonary Tuberculosis and in a Simulated Model of Dormancy |
| title_sort | differential culturability of mycobacterium tuberculosis in culture negative sputum of patients with pulmonary tuberculosis and in a simulated model of dormancy |
| topic | tuberculosis bacteriological diagnosis resuscitation Mycobacteirum tuberculosis clinical isolate organism sputum samples |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2019.02381/full |
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