Isothermal DNA amplification coupled to Au-nanoprobes for detection of mutations associated to Rifampicin resistance in Mycobacterium tuberculosis

Journal of Nanobiotechnology

Short Communication

Pedro Pedrosa¹  Pedro V Baptista¹  Bruno Veigas²  Isabel Couto³  Miguel Viveiros⁴ 

[1] Departamento de Ciências da Vida, Nanotheranostics@CIGMH, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Campus de Caparica, 2829-516, Caparica, Portugal;Departamento de Ciências da Vida, Nanotheranostics@CIGMH, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Campus de Caparica, 2829-516, Caparica, Portugal;Departamento de Ciência dos Materiais, CENIMAT/I3N, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Caparica, Portugal;Grupo de Micobactérias, Unidade Microbiologia Médica, Universidade Nova de Lisboa (IHMT/UNL), Lisboa, Portugal;CREM, Centro de Recursos Microbiológicos, Universidade Nova de Lisboa, Lisboa, Portugal;Grupo de Micobactérias, Unidade Microbiologia Médica, Universidade Nova de Lisboa (IHMT/UNL), Lisboa, Portugal;Centro de Malária e Outras Doenças Tropicais, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa (IHMT/UNL), Lisboa, Portugal;

关键词: MDRTB;    Nanodiagnostics;    LAMP;    PCR;    Gold nanoparticles;    Tuberculosis;    DNA isothermal amplification;    Rifampicin;   

DOI  :  10.1186/1477-3155-11-38

 received in 2013-10-15, accepted in 2013-11-22,  发布年份 2013

来源: Springer

【 摘 要 】

BackgroundTuberculosis accounted for 8.7 million new cases in 2011 and continues to be one of the leading human infectious diseases. Burdensome is the increasing rate of multi-drug resistant tuberculosis (MDRTB) and the difficulties created for treatment and public health control programs, especially in developing countries. Resistance to rifampicin (RIF), a first line antibiotic, is commonly associated with point mutations within the rpoB gene of Mycobacterium tuberculosis (Mtb) whose detection is considered the best early molecular predictor for MDRTB. Gold nanoparticles functionalized with thiol-modified oligonucleotides (Au-nanoprobes) have shown the potential to provide a rapid and sensitive detection method for Mtb and single base alterations associated with antibiotic resistance, namely in rpoB gene associated to RIF resistance.ResultsWe developed a strategy based on the isothermal amplification of sample DNA (LAMP) coupled to specific Au-nanoprobes capable of identifying members of the Mtb complex (MTBC) and discriminating specific mutations within the rpoB gene. Integration of LAMP and Au-nanoprobe assay allowed to detect MTBC member and identify mutations linked to RIF resistance. A total of 12 biological samples were tested and a 100% specificity and sensitivity was attained.ConclusionsThere is an increasing demand for simple, fast and cheap methods for the molecular identification of Mtb and for the detection of molecular tags associated to drug resistance suitable for use at point-of-need. Here we describe such a method, that as the potential to get molecular diagnostic of tuberculosis to remote environments.

【 授权许可】

Unknown   
© Veigas et al.; licensee BioMed Central Ltd. 2013. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.