title: A rapid and simple method for identifying Mycobacterium tuberculosis W-Beijing strains based on detection of a unique mutation in Rv0927c by PCR-SSCP.
abstract: Recently we have found that W-Beijing Mycobacterium tuberculosis strains have a unique in-frame trinucleotide (AGC) deletion at position 421 of Rv0927c and a -127G-->A mutation in Rv0927c-pstS3 intergenic region. Based on detecting the 421 trinucleotide deletion of these two mutations which can alter the ssDNA conformation more extensively than the other, we developed a PCR-SSCP method for rapid identification of W-Beijing strains among non-Beijing strains. Altogether, 104 clinical isolates were analyzed, including 68 W-Beijing strains and 36 non-Beijing strains. We found that PCR-SSCP successfully differentiated all the W-Beijing strains from the non-Beijing strains. In addition, we unexpectedly discovered that SDS-PAGE protein gels had better resolving power than conventional TBE polyacrylamide gel in detecting the AGC deletion mutation in the SSCP analysis.
authors.affiliation: Shanghai University of Traditional Chinese Medicine, Shanghai 201203, People's Republic of China.
authors.forename: Xin
authors.initials: X
authors.lastname: Jiang
authors.forename: Chanyi
authors.initials: C
authors.lastname: Lu
authors.forename: Feng
authors.initials: F
authors.lastname: Gao
authors.forename: Feifei
authors.initials: F
authors.lastname: Wang
authors.forename: Wenhong
authors.initials: W
authors.lastname: Zhang
authors.forename: Isabel
authors.initials: I
authors.lastname: Portugal
authors.forename: Peng
authors.initials: P
authors.lastname: Xu
authors.forename: Honghai
authors.initials: H
authors.lastname: Wang
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D018807:Polymorphism, Single-Stranded Conformational; D012680:Sensitivity and Specificity; D017384:Sequence Deletion
chemical_list: D001426:Bacterial Proteins