title: A homogeneous signal-on strategy for the detection of rpoB genes of Mycobacterium tuberculosis based on electrochemiluminescent graphene oxide and ferrocene quenching.
abstract: Tuberculosis (TB) remains one of the leading causes of morbidity and mortality all over the world and multidrug resistance TB (MDR-TB) pose a serious threat to the TB control and represent an increasing public health problem. In this work, we report a homogeneous signal-on electrochemiluminescence (ECL) DNA sensor for the sensitive and specific detection of rpoB genes of MDR-TB by using ruthenium(II) complex functionalized graphene oxide (Ru-GO) as suspension sensing interface and ferrocene-labeled ssDNA (Fc-ssDNA) as ECL intensity controller. The ECL of Ru-GO could be effectively quenched by Fc-ssDNA absorbed on the Ru-GO nanosheets. The Ru-GO has good discrimination ability over ssDNA and dsDNA. Mutant ssDNA target responsible for the drug resistant tuberculosis can hybridize with Fc-ssDNA and release Fc-ssDNA from Ru-GO surface, leading to the recovery of ECL. Mutant ssDNA target can be detected in a range from 0.1 to 100 nM with a detection limit of 0.04 nM. The proposed protocol is sensitive, specific, simple, time-saving and polymerase chain reaction free without complicated immobilization, separation and washing steps, which creates a simple but valuable tool for facilitating fast and accurate detection of disease related specific sequences or gene mutations.
authors.affiliation: CAS Key Laboratory of Soft Matter Chemistry, Department of Chemistry, University of Science and Technology of China , 96 Jinzhai Road, Hefei, Anhui 230026, P. R. China.
authors.forename: Fang
authors.initials: F
authors.lastname: Li
authors.forename: Yuqi
authors.initials: Y
authors.lastname: Yu
authors.forename: Qi
authors.initials: Q
authors.lastname: Li
authors.forename: Ming
authors.initials: M
authors.lastname: Zhou
authors.forename: Hua
authors.initials: H
authors.lastname: Cui
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D003062:Codon / Q000235:genetics; D004277:DNA, Single-Stranded / Q000737:chemistry; D012321:DNA-Directed RNA Polymerases; D018432:Drug Resistance, Multiple / Q000235:genetics; D004563:Electrochemistry; D005296:Ferrous Compounds / Q000737:chemistry*; D006108:Graphite / Q000737:chemistry*; D008163:Luminescent Measurements; D000075163:Metallocenes; D008958:Models, Molecular; D008968:Molecular Conformation; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics*; D010087:Oxides / Q000737:chemistry*; D020641:Polymorphism, Single Nucleotide; D012428:Ruthenium / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D003062:Codon; D004277:DNA, Single-Stranded; D005296:Ferrous Compounds; D000075163:Metallocenes; D010087:Oxides; C505805:rpoB protein, Mycobacterium tuberculosis; D006108:Graphite; D012428:Ruthenium; D012321:DNA-Directed RNA Polymerases; C004998:ferrocene