Gold nanoprobes for multi loci assessment of multi-drug resistant tuberculosis.
Tuberculosis, still one of the leading human infectious diseases, reported 8.7 million new cases in 2011 alone. Also, the increasing rate of multidrug-resistant tuberculosis (MDRTB) and its treatment difficulties pose a serious public health threat especially in developing countries. Resistance to isoniazid and rifampicin, first line antibiotics, is commonly associated with point mutations in katG, inhA and rpoB genes of Mycobacterium tuberculosis complex (MTBC). Therefore, the development of cheap, fast and simple molecular methods to assess susceptibility profiles would have a huge impact in the capacity of early diagnosis and treatment of MDRTB. Gold nanoparticles functionalized with thiol-modified oligonucleotides (Au-nanoprobes) have shown the potential to provide a rapid and sensitive detection method for MTBC and single base mutations associated with antibiotic resistance, namely the characterization of the three most relevant codons in rpoB gene associated to rifampicin resistance. Here we extend the Au-nanoprobe approach towards discriminating specific mutations within inhA and rpoB genes in PCR amplified DNA from isolates. Using a multiplex PCR reaction for these two genes, it is possible to assess both loci in parallel, and extend the potential of the Au-nanoprobe method to MDRTB molecular characterization with special application in the most frequent Portuguese genotypes.
CIGMH, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Caparica, Portugal.
Pedro
P
Pedrosa
CIGMH, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Caparica, Portugal; CENIMAT/I3N, Departamento de Ciência dos Materiais, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Caparica, Portugal.
Bruno
B
Veigas
Grupo de Micobactérias, Unidade Microbiologia Médica, Universidade Nova de Lisboa (IHMT/UNL), Portugal; Unidade de Parasitologia e Microbiologia Médicas, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa (IHMT/UNL), Portugal.
Diana
D
Machado
Grupo de Micobactérias, Unidade Microbiologia Médica, Universidade Nova de Lisboa (IHMT/UNL), Portugal; CREM, Centro de Recursos Microbiológicos, Universidade Nova de Lisboa, Portugal.
Isabel
I
Couto
Grupo de Micobactérias, Unidade Microbiologia Médica, Universidade Nova de Lisboa (IHMT/UNL), Portugal; Centro de Malária e Outras Doenças Tropicais, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa (IHMT/UNL), Portugal.
Miguel
M
Viveiros
CIGMH, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Caparica, Portugal. Electronic address: pmvb@fct.unl.pt.
Pedro V
PV
Baptista
D000995:Antitubercular Agents / Q000627:therapeutic use; D001426:Bacterial Proteins / Q000235:genetics; D012321:DNA-Directed RNA Polymerases; D042241:Early Diagnosis; D006046:Gold; D006801:Humans; D007538:Isoniazid / Q000627:therapeutic use; D053768:Metal Nanoparticles; D060885:Multiplex Polymerase Chain Reaction; D010088:Oxidoreductases / Q000235:genetics; D017354:Point Mutation / Q000235:genetics; D019095:Point-of-Care Systems; D012293:Rifampin / Q000627:therapeutic use; D018088:Tuberculosis, Multidrug-Resistant / Q000175:diagnosis* / Q000188:drug therapy / Q000235:genetics
D000995:Antitubercular Agents; D001426:Bacterial Proteins; C505805:rpoB protein, Mycobacterium tuberculosis; D006046:Gold; D010088:Oxidoreductases; C085257:InhA protein, Mycobacterium; D012321:DNA-Directed RNA Polymerases; D007538:Isoniazid; D012293:Rifampin
Gold nanoparticles; MDRTB; Nanodiagnostics; Tuberculosis; inhA; rpoB