Fluoromycobacteriophages can detect viable Mycobacterium tuberculosis and determine phenotypic rifampicin resistance in 3-5 days from sputum collection
The World Health Organization (WHO) estimates that 40% of tuberculosis (TB) cases are not diagnosed and treated correctly. Even though there are several diagnostic tests available in the market, rapid, easy, inexpensive detection, and drug susceptibility testing (DST) of Mycobacterium tuberculosis i...
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| Acceso en línea: | http://hdl.handle.net/20.500.12110/paper_1664302X_v9_nJUL_p_Rondon |
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todo:paper_1664302X_v9_nJUL_p_Rondon2023-10-03T16:29:07Z Fluoromycobacteriophages can detect viable Mycobacterium tuberculosis and determine phenotypic rifampicin resistance in 3-5 days from sputum collection Rondón, L. Urdániz, E. Latini, C. Payaslian, F. Matteo, M. Sosa, E.J. Do Porto, D.F. Turjanski, A.G. Nemirovsky, S. Hatfull, G.F. Poggi, S. Piuri, M. Diagnosis Drug susceptibility testing Fluoromycobacteriophages MCherrybombΦ Tuberculosis amikacin ethambutol isoniazid kanamycin ofloxacin rifampicin streptomycin acid fast bacterium adult antibiotic resistance antibiotic sensitivity antibiotic therapy Article atypical Mycobacterium bacterium culture bioinformatics clinical protocol comparative study diagnostic test accuracy study DNA isolation drug sensitivity female fluorescence microscopy fluoromycobacteriophage human male microbiological examination middle income country mycobacteriophage Mycobacterium tuberculosis next generation sequencing patient monitoring phenotype predictive value sensitivity and specificity sputum analysis transgender treatment response tuberculosis whole genome sequencing The World Health Organization (WHO) estimates that 40% of tuberculosis (TB) cases are not diagnosed and treated correctly. Even though there are several diagnostic tests available in the market, rapid, easy, inexpensive detection, and drug susceptibility testing (DST) of Mycobacterium tuberculosis is still of critical importance specially in low and middle-income countries with high incidence of the disease. In this work, we have developed a microscopy-based methodology using the reporter mycobacteriophage mCherrybombΦ for detection of Mycobacterium spp. and phenotypic determination of rifampicin resistance within just days from sputum sample collection. Fluoromycobacteriophage methodology is compatible with regularly used protocols in clinical laboratories for TB diagnosis and paraformaldehyde fixation after infection reduces biohazard risks with sample analysis by fluorescence microscopy. We have also set up conditions for discrimination between M. tuberculosis complex (MTBC) and non-tuberculous mycobacteria (NTM) strains by addition of p-nitrobenzoic acid (PNB) during the assay. Using clinical isolates of pre-XDR and XDR-TB strains from this study, we tested mCherrybombΦ for extended DST and we compared the antibiotic resistance profile with those predicted by whole genome sequencing. Our results emphasize the utility of a phenotypic test for M. tuberculosis extended DST. The many attributes of mCherrybombΦ suggests this could be a useful component of clinical microbiological laboratories for TB diagnosis and since only viable cells are detected this could be a useful tool for monitoring patient response to treatment. © 2018 Rondón, Urdániz, Latini, Payaslian, Matteo, Sosa, Do Porto, Turjanski, Nemirovsky, Hatfull, Poggi and Piuri. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_1664302X_v9_nJUL_p_Rondon |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Diagnosis Drug susceptibility testing Fluoromycobacteriophages MCherrybombΦ Tuberculosis amikacin ethambutol isoniazid kanamycin ofloxacin rifampicin streptomycin acid fast bacterium adult antibiotic resistance antibiotic sensitivity antibiotic therapy Article atypical Mycobacterium bacterium culture bioinformatics clinical protocol comparative study diagnostic test accuracy study DNA isolation drug sensitivity female fluorescence microscopy fluoromycobacteriophage human male microbiological examination middle income country mycobacteriophage Mycobacterium tuberculosis next generation sequencing patient monitoring phenotype predictive value sensitivity and specificity sputum analysis transgender treatment response tuberculosis whole genome sequencing |
| spellingShingle |
Diagnosis Drug susceptibility testing Fluoromycobacteriophages MCherrybombΦ Tuberculosis amikacin ethambutol isoniazid kanamycin ofloxacin rifampicin streptomycin acid fast bacterium adult antibiotic resistance antibiotic sensitivity antibiotic therapy Article atypical Mycobacterium bacterium culture bioinformatics clinical protocol comparative study diagnostic test accuracy study DNA isolation drug sensitivity female fluorescence microscopy fluoromycobacteriophage human male microbiological examination middle income country mycobacteriophage Mycobacterium tuberculosis next generation sequencing patient monitoring phenotype predictive value sensitivity and specificity sputum analysis transgender treatment response tuberculosis whole genome sequencing Rondón, L. Urdániz, E. Latini, C. Payaslian, F. Matteo, M. Sosa, E.J. Do Porto, D.F. Turjanski, A.G. Nemirovsky, S. Hatfull, G.F. Poggi, S. Piuri, M. Fluoromycobacteriophages can detect viable Mycobacterium tuberculosis and determine phenotypic rifampicin resistance in 3-5 days from sputum collection |
| topic_facet |
Diagnosis Drug susceptibility testing Fluoromycobacteriophages MCherrybombΦ Tuberculosis amikacin ethambutol isoniazid kanamycin ofloxacin rifampicin streptomycin acid fast bacterium adult antibiotic resistance antibiotic sensitivity antibiotic therapy Article atypical Mycobacterium bacterium culture bioinformatics clinical protocol comparative study diagnostic test accuracy study DNA isolation drug sensitivity female fluorescence microscopy fluoromycobacteriophage human male microbiological examination middle income country mycobacteriophage Mycobacterium tuberculosis next generation sequencing patient monitoring phenotype predictive value sensitivity and specificity sputum analysis transgender treatment response tuberculosis whole genome sequencing |
| description |
The World Health Organization (WHO) estimates that 40% of tuberculosis (TB) cases are not diagnosed and treated correctly. Even though there are several diagnostic tests available in the market, rapid, easy, inexpensive detection, and drug susceptibility testing (DST) of Mycobacterium tuberculosis is still of critical importance specially in low and middle-income countries with high incidence of the disease. In this work, we have developed a microscopy-based methodology using the reporter mycobacteriophage mCherrybombΦ for detection of Mycobacterium spp. and phenotypic determination of rifampicin resistance within just days from sputum sample collection. Fluoromycobacteriophage methodology is compatible with regularly used protocols in clinical laboratories for TB diagnosis and paraformaldehyde fixation after infection reduces biohazard risks with sample analysis by fluorescence microscopy. We have also set up conditions for discrimination between M. tuberculosis complex (MTBC) and non-tuberculous mycobacteria (NTM) strains by addition of p-nitrobenzoic acid (PNB) during the assay. Using clinical isolates of pre-XDR and XDR-TB strains from this study, we tested mCherrybombΦ for extended DST and we compared the antibiotic resistance profile with those predicted by whole genome sequencing. Our results emphasize the utility of a phenotypic test for M. tuberculosis extended DST. The many attributes of mCherrybombΦ suggests this could be a useful component of clinical microbiological laboratories for TB diagnosis and since only viable cells are detected this could be a useful tool for monitoring patient response to treatment. © 2018 Rondón, Urdániz, Latini, Payaslian, Matteo, Sosa, Do Porto, Turjanski, Nemirovsky, Hatfull, Poggi and Piuri. |
| format |
JOUR |
| author |
Rondón, L. Urdániz, E. Latini, C. Payaslian, F. Matteo, M. Sosa, E.J. Do Porto, D.F. Turjanski, A.G. Nemirovsky, S. Hatfull, G.F. Poggi, S. Piuri, M. |
| author_facet |
Rondón, L. Urdániz, E. Latini, C. Payaslian, F. Matteo, M. Sosa, E.J. Do Porto, D.F. Turjanski, A.G. Nemirovsky, S. Hatfull, G.F. Poggi, S. Piuri, M. |
| author_sort |
Rondón, L. |
| title |
Fluoromycobacteriophages can detect viable Mycobacterium tuberculosis and determine phenotypic rifampicin resistance in 3-5 days from sputum collection |
| title_short |
Fluoromycobacteriophages can detect viable Mycobacterium tuberculosis and determine phenotypic rifampicin resistance in 3-5 days from sputum collection |
| title_full |
Fluoromycobacteriophages can detect viable Mycobacterium tuberculosis and determine phenotypic rifampicin resistance in 3-5 days from sputum collection |
| title_fullStr |
Fluoromycobacteriophages can detect viable Mycobacterium tuberculosis and determine phenotypic rifampicin resistance in 3-5 days from sputum collection |
| title_full_unstemmed |
Fluoromycobacteriophages can detect viable Mycobacterium tuberculosis and determine phenotypic rifampicin resistance in 3-5 days from sputum collection |
| title_sort |
fluoromycobacteriophages can detect viable mycobacterium tuberculosis and determine phenotypic rifampicin resistance in 3-5 days from sputum collection |
| url |
http://hdl.handle.net/20.500.12110/paper_1664302X_v9_nJUL_p_Rondon |
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