title: Au-nanoprobes for detection of SNPs associated with antibiotic resistance in Mycobacterium tuberculosis.
abstract: Tuberculosis (TB) is one of the leading causes of infection in humans, causing high morbility and mortality all over the world. The rate of new cases of multidrug resistant tuberculosis (MDRTB) continues to increase, and since these infections are very difficult to manage, they constitute a serious health problem. In most cases, drug resistance in Mycobacterium tuberculosis has been related to mutations in several loci within the pathogen's genome. The development of fast, cheap and simple screening methodologies would be of paramount relevance for the early detection of these mutations, essential for the timely and effective diagnosis and management of MDRTB patients. The use of gold nanoparticles derivatized with thiol-modified oligonucleotides (Au-nanoprobes) has led to new approaches in molecular diagnostics. Based on the differential non-cross-linking aggregation of Au-nanoprobes, we were able to develop a colorimetric method for the detection of specific sequences and to apply this approach to pathogen identification and single base mutations/single nucleotide polymorphisms (SNP) discrimination. Here we report on the development of Au-nanoprobes for the specific identification of SNPs within the beta subunit of the RNA polymerase (rpoB locus), responsible for resistance to rifampicin in over 95% of rifampicin resistant M. tuberculosis strains.
authors.affiliation: CIGMH, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Caparica, Portugal.
authors.forename: Bruno
authors.initials: B
authors.lastname: Veigas
authors.forename: Diana
authors.initials: D
authors.lastname: Machado
authors.forename: João
authors.initials: J
authors.lastname: Perdigão
authors.forename: Isabel
authors.initials: I
authors.lastname: Portugal
authors.forename: Isabel
authors.initials: I
authors.lastname: Couto
authors.forename: Miguel
authors.initials: M
authors.lastname: Viveiros
authors.forename: Pedro V
authors.initials: PV
authors.lastname: Baptista
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D004252:DNA Mutational Analysis; D004352:Drug Resistance, Microbial / Q000187:drug effects / Q000235:genetics*; D005798:Genes, Bacterial / Q000235:genetics; D006046:Gold / Q000737:chemistry*; D006801:Humans; D053768:Metal Nanoparticles / Q000737:chemistry*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D012293:Rifampin / Q000494:pharmacology
chemical_list: D001426:Bacterial Proteins; D006046:Gold; D012293:Rifampin