title: Visual detection of rpoB mutations in rifampin-resistant Mycobacterium tuberculosis strains by use of an asymmetrically split peroxidase DNAzyme.
abstract: Multidrug-resistant Mycobacterium tuberculosis is resistant to two first-line antituberculosis drugs, isoniazid and rifampin, resulting in the relapse of tuberculosis. M. tuberculosis grows very slowly, and thus traditional examination methods take time to test its drug resistance and cannot meet clinical needs. The use of a DNA probe makes it possible to test rifampin resistance.  We developed an asymmetrical split-assembly DNA peroxidase assay to detect drug-resistant mutation of rifampin-resistant M. tuberculosis in the rpoB gene rapidly and visibly. A new strategy was also designed to eliminate the adverse effects caused by the complicated secondary structure of the target DNA and to improve the efficiency of the probes. This detection system consists of five group detections, covers rifampin-resistant determination region of the rpoB gene, and tests 40 kinds of mutations, including the most common mutations at codons 531 and 526. Every group detection or individual mutant allele detection can distinguish corresponding mutant DNA sequences from the wild-type DNA sequences.
authors.affiliation: State Key Laboratory of Virology, School of Medicine, Wuhan University, Wuhan, People’s Republic of China.
authors.forename: Minggang
authors.initials: M
authors.lastname: Deng
authors.forename: Shuo
authors.initials: S
authors.lastname: Feng
authors.forename: Fengling
authors.initials: F
authors.lastname: Luo
authors.forename: Shaoru
authors.initials: S
authors.lastname: Wang
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Sun
authors.forename: Xiang
authors.initials: X
authors.lastname: Zhou
authors.forename: Xiao-Lian
authors.initials: XL
authors.lastname: Zhang
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D021881:DNA, Catalytic / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases; D024881:Drug Resistance, Bacterial; D006801:Humans; D020125:Mutation, Missense; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification; D009195:Peroxidase / Q000378:metabolism; D012293:Rifampin / Q000494:pharmacology*; D018088:Tuberculosis, Multidrug-Resistant / Q000175:diagnosis / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins; D021881:DNA, Catalytic; C505805:rpoB protein, Mycobacterium tuberculosis; D009195:Peroxidase; D012321:DNA-Directed RNA Polymerases; D012293:Rifampin