Gold nanoprobe assay for the identification of mycobacteria of the Mycobacterium tuberculosis complex

• Published in: Clinical microbiology and infection Vol. 16; no. 9; pp. 1464 - 1469
• Main Authors: Costa, P., Amaro, A., Botelho, A., Inácio, J., Baptista, P.V.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Elsevier Ltd 01.09.2010; Blackwell Publishing Ltd; Wiley-Blackwell; Elsevier Limited
• Subjects: Bacteriological Techniques - methods; Biological and medical sciences; Colorimetry; Colorimetry - methods; Differentiation; DNA Gyrase - genetics; DNA topoisomerase; Gold; Gold nanoprobes; Humans; Infectious diseases; Medical sciences; Metal Nanoparticles; Molecular Diagnostic Techniques - methods; Mycobacterium bovis; Mycobacterium bovis - classification; Mycobacterium bovis - isolation & purification; Mycobacterium tuberculosis; Mycobacterium tuberculosis - classification; Mycobacterium tuberculosis - isolation & purification; Mycobacterium tuberculosis complex; Nucleic Acid Hybridization; Nucleotide sequence; Phylogeny; Sensitivity and Specificity; Tuberculosis; Tuberculosis - diagnosis; Tuberculosis - microbiology; Mycobacterium bovis; Mycobacterium tuberculosis complex; Gold nanoprobes; Infection; Mycobacterium tuberculosis; Mycobacteriales; Mycobacteriaceae; Bacteria; Actinomycetes; Identification
• ISSN: 1198-743X; 1469-0691
• DOI: 10.1111/j.1469-0691.2010.03120.x

Members of the Mycobacterium tuberculosis complex (MTC) are causative agents of human and animal tuberculosis. This complex encompasses several phylogenetically related species, including M. tuberculosis, the main aetiological agent of human tuberculosis, and Mycobacterium bovis, the causative agent of bovine tuberculosis, a relevant worldwide zoonosis. Clear epidemiological evaluation of appropriate and effective treatment requires unambiguous differentiation between MTC members. Routine diagnosis has been increasingly relying on the molecular identification of MTC members. In the present study, we report the use of a gold nanoparticle-based approach for the sensitive, specific and fast identification of MTC and for the differentiation of M. bovis and M. tuberculosis using the gyrB locus as target. This gold nanoprobe strategy relies on the colorimetric differentiation of specific DNA sequences based on differential aggregation profiles in the presence or absence of specific target hybridization. Three nanoprobes were designed and successfully used for the specific identification of members of MTC, M. bovis and M. tuberculosis.