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.
['Pedrosa|Pedro|P|', 'Veigas|Bruno|B|', 'Machado|Diana|D|', 'Couto|Isabel|I|', 'Viveiros|Miguel|M|', 'Baptista|Pedro V|PV|']
D000995:Antitubercular Agents D001426:Bacterial Proteins D012321:DNA-Directed RNA Polymerases D042241:Early Diagnosis D006046:Gold D006801:Humans D007538:Isoniazid D053768:Metal Nanoparticles D060885:Multiplex Polymerase Chain Reaction D010088:Oxidoreductases D017354:Point Mutation D019095:Point-of-Care Systems D012293:Rifampin D018088:Tuberculosis, Multidrug-Resistant
Tuberculosis MDRTB inhA rpoB Gold nanoparticles Nanodiagnostics