Sulfatase-activated fluorophores for rapid discrimination of mycobacterial species and strains

Most current diagnostic tests for tuberculosis do not reveal the species or strain of pathogen causing pulmonary infection, which can lead to inappropriate treatment regimens and the spread of disease. Here, we report an assay for mycobacterial strain assignment based on genetically conserved mycoba...

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Published inProceedings of the National Academy of Sciences - PNAS Vol. 110; no. 32; pp. 12911 - 12916
Main Authors Beatty, Kimberly E., Williams, Monique, Carlson, Brian L., Swarts, Benjamin M., Warren, Robin M., van Helden, Paul D., Bertozzi, Carolyn R.
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences 06.08.2013
National Acad Sciences
Subjects
Acridones - chemistry
Animals
Bacteria
Bacterial Proteins - metabolism
Biological Sciences
Cell Line
Cell Line, Tumor
Cercopithecus aethiops
CHO Cells
COS Cells
Cricetinae
Cricetulus
Electrophoresis - methods
Enzymes
Esters
Fluorescence
Fluorescent Dyes - chemistry
Gels
Genomes
HEK293 Cells
Humans
Hydrolysis
Infectious diseases
Jurkat Cells
Medical treatment
Molecular Structure
Mycobacterium
Mycobacterium - classification
Mycobacterium - enzymology
Mycobacterium - metabolism
Mycobacterium tuberculosis
Mycobacterium tuberculosis - enzymology
Mycobacterium tuberculosis - metabolism
Physical Sciences
Proteins
Pulmonary tuberculosis
Species Specificity
Substrate Specificity
Sulfatases - metabolism
Sulfates
Sulfates - chemistry
Sulfates - metabolism
Sulfuric Acid Esters - chemistry
Tuberculosis
Tuberculosis - diagnosis
Tuberculosis - microbiology
enzyme assay
Mycobacterium tuberculosis
chemical biology
fluorescence
hydrolase
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Summary:Most current diagnostic tests for tuberculosis do not reveal the species or strain of pathogen causing pulmonary infection, which can lead to inappropriate treatment regimens and the spread of disease. Here, we report an assay for mycobacterial strain assignment based on genetically conserved mycobacterial sulfatases. We developed a sulfatase-activated probe, 7-hydroxy-9H-(1,3-dichloro-9,9-dimethylacridin-2-one)–sulfate, that detects enzyme activity in native protein gels, allowing the rapid detection of sulfatases in mycobacterial lysates. This assay revealed that mycobacterial strains have distinct sulfatase fingerprints that can be used to judge both the species and lineage. Our results demonstrate the potential of enzyme-activated probes for rapid pathogen discrimination for infectious diseases.
Bibliography:http://dx.doi.org/10.1073/pnas.1222041110
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1Present address: Department of Physiology and Pharmacology, Oregon Health and Science University, Portland, OR 97239.
Edited by Laura L. Kiessling, University of Wisconsin-Madison, Madison, WI, and approved June 7, 2013 (received for review January 4, 2013)
Author contributions: K.E.B. and C.R.B. designed research; K.E.B., M.W., B.L.C., and B.M.S. performed research; K.E.B., M.W., B.L.C., B.M.S., R.M.W., P.D.v.H., and C.R.B. contributed new reagents/analytic tools; K.E.B., M.W., B.L.C., B.M.S., R.M.W., P.D.v.H., and C.R.B. analyzed data; and K.E.B. and C.R.B. wrote the paper.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.1222041110