title: Simple multiplex PCR assay for identification of Beijing family Mycobacterium tuberculosis isolates with a lineage-specific mutation in Rv0679c.
abstract: The Beijing genotype of Mycobacterium tuberculosis is known to be a worldwide epidemic clade. It is suggested to be a possibly resistant clone against BCG vaccination and is also suggested to be highly pathogenic and prone to becoming drug resistant. Thus, monitoring the prevalence of this lineage seems to be important for the proper control of tuberculosis. The Rv0679c protein of M. tuberculosis has been predicted to be one of the outer membrane proteins and is suggested to contribute to host cell invasion. Here, we conducted a sequence analysis of the Rv0679c gene using clinical isolates and found that a single nucleotide polymorphism, C to G at position 426, can be observed only in the isolates that are identified as members of the Beijing genotype family. Here, we developed a simple multiplex PCR assay to detect this point mutation and applied it to 619 clinical isolates. The method successfully distinguished Beijing lineage clones from non-Beijing strains with 100% accuracy. This simple, quick, and cost-effective multiplex PCR assay can be used for a survey or for monitoring the prevalence of Beijing genotype M. tuberculosis strains.
authors.affiliation: Division of Global Epidemiology, Hokkaido University Research Center for Zoonosis Control, Sapporo, Hokkaido, Japan.
authors.forename: Chie
authors.initials: C
authors.lastname: Nakajima
authors.forename: Aki
authors.initials: A
authors.lastname: Tamaru
authors.forename: Zeaur
authors.initials: Z
authors.lastname: Rahim
authors.forename: Ajay
authors.initials: A
authors.lastname: Poudel
authors.forename: Bhagwan
authors.initials: B
authors.lastname: Maharjan
authors.lastname: Khin Saw Aye
authors.forename: Hong
authors.initials: H
authors.lastname: Ling
authors.forename: Toshio
authors.initials: T
authors.lastname: Hattori
authors.forename: Tomotada
authors.initials: T
authors.lastname: Iwamoto
authors.forename: Yukari
authors.initials: Y
authors.lastname: Fukushima
authors.forename: Haruka
authors.initials: H
authors.lastname: Suzuki
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Suzuki
authors.forename: Takashi
authors.initials: T
authors.lastname: Matsuba
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D005838:Genotype; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D058889:Molecular Typing / Q000379:methods; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D050505:Mutant Proteins / Q000235:genetics; D009169:Mycobacterium tuberculosis / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D020641:Polymorphism, Single Nucleotide; D014376:Tuberculosis / Q000175:diagnosis / Q000382:microbiology*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D050505:Mutant Proteins