title: Multiplex strand displacement amplification (SDA) and detection of DNA sequences from Mycobacterium tuberculosis and other mycobacteria.
abstract: Strand Displacement Amplification (SDA) is an isothermal, in vitro method of amplifying a DNA target sequence prior to detection [Walker et al (1992) Nucleic Acids Res., 20, 1691-1693]. Here we describe a multiplex form of SDA that allows two target sequences and an internal amplification control to be co-amplified by a single pair of primers after common priming sequences are spontaneously appended to the ends of target fragments. Multiplex SDA operates at a single temperature, under the same simple protocol previously developed for single-target SDA. We applied multiplex SDA to co-amplification of a target sequence (IS6110) that is specific to members of the Mycobacterium tuberculosis-complex and a target (16S ribosomal gene) that is common to most clinically relevant species of mycobacteria. Both targets are amplified 10(8)-fold during a 2 hour, single temperature incubation. The relative sensitivity of the system was evaluated across a number of clinically relevant mycobacteria and checked for crossreactivity against organisms that are closely related to mycobacteria.
authors.affiliation: Becton Dickinson Research Center, Research Triangle Park, NC 27709-2016.
authors.forename: G T
authors.initials: GT
authors.lastname: Walker
authors.forename: J G
authors.initials: JG
authors.lastname: Nadeau
authors.forename: P A
authors.initials: PA
authors.lastname: Spears
authors.forename: J L
authors.initials: JL
authors.lastname: Schram
authors.forename: C M
authors.initials: CM
authors.lastname: Nycz
authors.forename: D D
authors.initials: DD
authors.lastname: Shank
mesh_terms: D001483:Base Sequence; D004269:DNA, Bacterial / Q000032:analysis*; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000235:genetics*; D009169:Mycobacterium tuberculosis / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques; D012045:Regulatory Sequences, Nucleic Acid
chemical_list: D004269:DNA, Bacterial