title: Confirmation of the presence of Mycobacterium tuberculosis and other mycobacteria in mycobacterial growth indicator tubes (MGIT) by multiplex strand displacement amplification.
abstract: Multiplex strand displacement amplification (mSDA) is capable of amplifying three distinct DNA sequences simultaneously. These include sequences present in most genera of mycobacteria, a sequence specific for Mycobacterium tuberculosis, and an internal control. mSDA was used to detect the presence of these target sequences in 154 (72 positive, 76 negative, and 6 failed) clinical specimens cultured in the mycobacterial growth indicator tube (MGIT) system. A wide variety of specimen types were processed and cultured. Once these cultures were deemed positive by MGIT fluorescence or were deemed negative after 8 weeks of incubation, MGIT culture aliquots were processed for mSDA analyses. A chemiluminescent microwell assay was used to detect the amplified products. The procedure was relatively simple and took less than 6 h to complete. The sensitivity of mSDA for detecting acid-fast bacilli was 96.4% compared to that of MGIT culture. Sensitivity and specificity were 97.2 and 96.1%, respectively, when all clinical criteria were considered. mSDA was shown to be a rapid and effective method for confirming the presence of M. tuberculosis and other mycobacteria in positive MGIT cultures.
authors.affiliation: Becton Dickinson Research Center, Research Triangle Park, North Carolina, USA.
authors.forename: F Z
authors.initials: FZ
authors.lastname: Badak
authors.forename: D L
authors.initials: DL
authors.lastname: Kiska
authors.forename: M
authors.initials: M
authors.lastname: O'Connell
authors.forename: C M
authors.initials: CM
authors.lastname: Nycz
authors.forename: C
authors.initials: C
authors.lastname: Hartley
authors.forename: S
authors.initials: S
authors.lastname: Setterquist
authors.forename: R L
authors.initials: RL
authors.lastname: Hopfer
mesh_terms: D015373:Bacterial Typing Techniques; D004269:DNA, Bacterial / Q000032:analysis*; D009161:Mycobacterium / Q000145:classification*; D009169:Mycobacterium tuberculosis / Q000145:classification* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA
chemical_list: D004269:DNA, Bacterial