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Rapid Tuberculosis Diagnosis Using Reporter Enzyme Fluorescence

Tuberculosis is the most frequent cause of death in humans from a single infectious agent. Due to low numbers of bacteria present in sputum during early infection, diagnosis does not usually occur until >3 to 4 months after symptoms develop. We created a new more sensitive diagnostic that can be...

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Autores principales: Sule, Preeti, Tilvawala, Ronak, Mustapha, Toriq, Hassounah, Hany, Noormohamed, Aneesa, Kundu, Suprateek, Graviss, Edward A., Walkup, Grant K., Kong, Ying, Cirillo, Jeffrey D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6879286/
https://www.ncbi.nlm.nih.gov/pubmed/31511338
http://dx.doi.org/10.1128/JCM.01462-19
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author Sule, Preeti
Tilvawala, Ronak
Mustapha, Toriq
Hassounah, Hany
Noormohamed, Aneesa
Kundu, Suprateek
Graviss, Edward A.
Walkup, Grant K.
Kong, Ying
Cirillo, Jeffrey D.
author_facet Sule, Preeti
Tilvawala, Ronak
Mustapha, Toriq
Hassounah, Hany
Noormohamed, Aneesa
Kundu, Suprateek
Graviss, Edward A.
Walkup, Grant K.
Kong, Ying
Cirillo, Jeffrey D.
author_sort Sule, Preeti
collection PubMed
description Tuberculosis is the most frequent cause of death in humans from a single infectious agent. Due to low numbers of bacteria present in sputum during early infection, diagnosis does not usually occur until >3 to 4 months after symptoms develop. We created a new more sensitive diagnostic that can be carried out in 10 min with no processing or technical expertise. This assay utilizes the Mycobacterium tuberculosis-specific biomarker BlaC in reporter enzyme fluorescence (REF) that has been optimized for clinical samples, designated REFtb, along with a more specific fluorogenic substrate, CDG-3. We report the first evaluation of clinical specimens with REFtb assays in comparison to the gold standards for tuberculosis diagnosis, culture and smear microscopy. REFtb assays allowed diagnosis of 160 patients from 16 different countries with a sensitivity of 89% for smear-positive, culture-positive samples and 88% for smear-negative, culture-positive samples with a specificity of 82%. The negative predictive value of REFtb for tuberculosis infection is 93%, and the positive predictive value is 79%. Overall, these data point toward the need for larger accuracy studies by third parties using a commercially available REFtb kit to determine whether incorporation of REFtb into the clinical toolbox for suspected tuberculosis patients would improve case identification. If results similar to our own can be obtained by all diagnostic laboratories, REFtb would allow proper treatment of more than 85% of patients that would be missed during their initial visit to a clinic using current diagnostic strategies, reducing the potential for further spread of disease.
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spelling pubmed-68792862019-12-03 Rapid Tuberculosis Diagnosis Using Reporter Enzyme Fluorescence Sule, Preeti Tilvawala, Ronak Mustapha, Toriq Hassounah, Hany Noormohamed, Aneesa Kundu, Suprateek Graviss, Edward A. Walkup, Grant K. Kong, Ying Cirillo, Jeffrey D. J Clin Microbiol Mycobacteriology and Aerobic Actinomycetes Tuberculosis is the most frequent cause of death in humans from a single infectious agent. Due to low numbers of bacteria present in sputum during early infection, diagnosis does not usually occur until >3 to 4 months after symptoms develop. We created a new more sensitive diagnostic that can be carried out in 10 min with no processing or technical expertise. This assay utilizes the Mycobacterium tuberculosis-specific biomarker BlaC in reporter enzyme fluorescence (REF) that has been optimized for clinical samples, designated REFtb, along with a more specific fluorogenic substrate, CDG-3. We report the first evaluation of clinical specimens with REFtb assays in comparison to the gold standards for tuberculosis diagnosis, culture and smear microscopy. REFtb assays allowed diagnosis of 160 patients from 16 different countries with a sensitivity of 89% for smear-positive, culture-positive samples and 88% for smear-negative, culture-positive samples with a specificity of 82%. The negative predictive value of REFtb for tuberculosis infection is 93%, and the positive predictive value is 79%. Overall, these data point toward the need for larger accuracy studies by third parties using a commercially available REFtb kit to determine whether incorporation of REFtb into the clinical toolbox for suspected tuberculosis patients would improve case identification. If results similar to our own can be obtained by all diagnostic laboratories, REFtb would allow proper treatment of more than 85% of patients that would be missed during their initial visit to a clinic using current diagnostic strategies, reducing the potential for further spread of disease. American Society for Microbiology 2019-11-22 /pmc/articles/PMC6879286/ /pubmed/31511338 http://dx.doi.org/10.1128/JCM.01462-19 Text en Copyright © 2019 Sule et al. https://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Mycobacteriology and Aerobic Actinomycetes
Sule, Preeti
Tilvawala, Ronak
Mustapha, Toriq
Hassounah, Hany
Noormohamed, Aneesa
Kundu, Suprateek
Graviss, Edward A.
Walkup, Grant K.
Kong, Ying
Cirillo, Jeffrey D.
Rapid Tuberculosis Diagnosis Using Reporter Enzyme Fluorescence
title Rapid Tuberculosis Diagnosis Using Reporter Enzyme Fluorescence
title_full Rapid Tuberculosis Diagnosis Using Reporter Enzyme Fluorescence
title_fullStr Rapid Tuberculosis Diagnosis Using Reporter Enzyme Fluorescence
title_full_unstemmed Rapid Tuberculosis Diagnosis Using Reporter Enzyme Fluorescence
title_short Rapid Tuberculosis Diagnosis Using Reporter Enzyme Fluorescence
title_sort rapid tuberculosis diagnosis using reporter enzyme fluorescence
topic Mycobacteriology and Aerobic Actinomycetes
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6879286/
https://www.ncbi.nlm.nih.gov/pubmed/31511338
http://dx.doi.org/10.1128/JCM.01462-19
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