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Molecular bacterial load assay (MBLA) concurs with culture on the NaOH-induced Mycobacterium tuberculosis loss of viability
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dc.contributor.author | Mtafya, Bariki | |
dc.contributor.author | Sabiiti, Wilber | |
dc.contributor.author | Sabi, Issa | |
dc.contributor.author | John, Joseph | |
dc.contributor.author | Sichone, Emanuel | |
dc.contributor.author | Ntinginya, Nyanda E. | |
dc.contributor.author | Gillespie, Stephen H. | |
dc.date.accessioned | 2019-10-23T23:36:57Z | |
dc.date.available | 2019-10-23T23:36:57Z | |
dc.date.issued | 2019-06-25 | |
dc.identifier | 258657292 | |
dc.identifier | 7da6c22f-5b79-4a17-a116-e4d25cc3c539 | |
dc.identifier | 000472800800019 | |
dc.identifier | 85068554734 | |
dc.identifier.citation | Mtafya , B , Sabiiti , W , Sabi , I , John , J , Sichone , E , Ntinginya , N E & Gillespie , S H 2019 , ' Molecular bacterial load assay (MBLA) concurs with culture on the NaOH-induced Mycobacterium tuberculosis loss of viability ' , Journal of Clinical Microbiology , vol. 57 , e01992-18 . https://doi.org/10.1128/JCM.01992-18 | en |
dc.identifier.issn | 0095-1137 | |
dc.identifier.other | ORCID: /0000-0001-6537-7712/work/58531611 | |
dc.identifier.other | ORCID: /0000-0002-4742-2791/work/60196329 | |
dc.identifier.uri | https://hdl.handle.net/10023/18753 | |
dc.description | This work was supported by the commonwealth studentship award for Bariki Mtafya at University of St Andrews in UK and European and Developing Countries Clinical Trials Partnership (EDCTP) through TWENDE and PanACEA II grants. | en |
dc.description.abstract | Effective methods to detect viable Mycobacterium tuberculosis (Mtb), the main causative agent of tuberculosis (TB) are urgently needed. To date, cultivation of Mtb is the gold standard which depends on initial sample processing with N-Acetyl-L-Cysteine/Sodium hydroxide (NALC/NaOH), chemicals that compromise Mtb viability and, consequently the performance of downstream tests. We applied culture and the novel Molecular bacterial load assay (MBLA) to measure the loss of Mtb viability following NALC/NaOH treatment of Mtb H37Rv pure culture and clinical sputa from pulmonary TB patients. Compared to untreated controls, NALC/NaOH treatment of Mtb, reduced MBLA detectable bacillary load (estimated colony forming units/milliliter (eCFU/mL) by 0.66±0.21log10- at 23°C (P=0.018) and 0.72±0.08log10- at 30°C (P=0.013). Likewise, NALC/NaOH treatment reduced viable count on solid culture by 0.84±0.02log10- at 23°C (P<0.001) and 0.85±0.01log10- CFU/mL at 30°C (P<0.001) respectively. The reduction in viable count was reflected by a corresponding increase in time to positivity of MGIT liquid culture, 1.2 days at 23°C (P<0.001), and 1.1 days at 30°C (P<0.001). This NaOH-induced Mtb viability loss was replicated in clinical sputum samples, with bacterial load dropping by 0.65±0.17log10 from 5.36±0.24log10- to 4.71±0.16log10- eCFU/mL for untreated and treated sputa respectively. Applying the Bowness et al model, revealed that the treated MGIT time to culture positivity of 142hrs was equivalent to 4.86±0.28log10CFU, consistent with MBLA-measured bacterial load. Our study confirms the contribution of NALC/NaOH treatment to loss of viable bacterial count. Tests that obviate the need of decontamination may offer alternative option for accurate detection of viable Mtb and treatment response monitoring. | |
dc.format.extent | 931468 | |
dc.language.iso | eng | |
dc.relation.ispartof | Journal of Clinical Microbiology | en |
dc.subject | QR Microbiology | en |
dc.subject | RA0421 Public health. Hygiene. Preventive Medicine | en |
dc.subject | NDAS | en |
dc.subject | SDG 3 - Good Health and Well-being | en |
dc.subject.lcc | QR | en |
dc.subject.lcc | RA0421 | en |
dc.title | Molecular bacterial load assay (MBLA) concurs with culture on the NaOH-induced Mycobacterium tuberculosis loss of viability | en |
dc.type | Journal article | en |
dc.contributor.institution | University of St Andrews. School of Medicine | en |
dc.contributor.institution | University of St Andrews. Infection and Global Health Division | en |
dc.contributor.institution | University of St Andrews. Sir James Mackenzie Institute for Early Diagnosis | en |
dc.contributor.institution | University of St Andrews. Centre for Biophotonics | en |
dc.contributor.institution | University of St Andrews. Global Health Implementation Group | en |
dc.contributor.institution | University of St Andrews. Gillespie Group | en |
dc.contributor.institution | University of St Andrews. Infection Group | en |
dc.contributor.institution | University of St Andrews. Biomedical Sciences Research Complex | en |
dc.identifier.doi | 10.1128/JCM.01992-18 | |
dc.description.status | Peer reviewed | en |
dc.date.embargoedUntil | 2019-10-24 |
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