title: High-resolution melting facilitates mutation screening of rpsL gene associated with streptomycin resistance in Mycobacterium tuberculosis.
abstract: Drug resistance remains a serious threat to tuberculosis control worldwide. As one of the important first-line antitubercular agents, resistance to streptomycin (SM) and its derivatives has increased in recent years and has become one of the characteristics of extensively drug-resistant tuberculosis (XDR-TB). A novel rapid assay to screen for rpsL gene mutations associated with SM resistance in Mycobacterium tuberculosis (M. tuberculosis), was developed using high-resolution melting (HRM) analysis. The HRM results of 134 SM-resistant clinical isolates and 20 SM-susceptible clinical isolates of M. tuberculosis for rpsL gene mutations were perfectly matched with those of DNA sequencing. SM resistance was highly associated with rpsL mutations in M. tuberculosis. HRM technique represented an inexpensive, highly sensitive and high-throughput method to facilitate the screening of large numbers of clinical samples for epidemiological studies of drug-resistance of M. tuberculosis, especially in developing countries.
authors.affiliation: State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Science, Fudan University, Shanghai 200433, China.
authors.forename: Feifei
authors.initials: F
authors.lastname: Wang
authors.forename: Hongbo
authors.initials: H
authors.lastname: Shen
authors.forename: Ming
authors.initials: M
authors.lastname: Guan
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.forename: Yun
authors.initials: Y
authors.lastname: Feng
authors.forename: Xinhua
authors.initials: X
authors.lastname: Weng
authors.forename: Honghai
authors.initials: H
authors.lastname: Wang
authors.forename: Wenhong
authors.initials: W
authors.lastname: Zhang
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods*; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics*; D009691:Nucleic Acid Denaturation; D015203:Reproducibility of Results; D012269:Ribosomal Proteins / Q000235:genetics*; D017422:Sequence Analysis, DNA; D013307:Streptomycin / Q000494:pharmacology; D014376:Tuberculosis / Q000382:microbiology
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D012269:Ribosomal Proteins; D013307:Streptomycin