title: Multiplex PCR assay specific for the multidrug-resistant strain W of Mycobacterium tuberculosis.
abstract: In 1991, a multidrug-resistant strain of Mycobacterium tuberculosis was isolated from eight people with tuberculosis at a state correctional facility in New York. This strain, which is designated strain W (IS6110 restriction fragment length polymorphism type 212072), was resistant to isoniazid, rifampin, ethambutol, streptomycin, kanamycin, ethionamide, and rifabutin. Since that outbreak, the W strain has been associated with outbreaks in five hospitals in the New York City area and is a continuing public health problem in the area. To be able to identify this strain rapidly, we developed a multiplex PCR assay which targets a direct repeat of IS6110 with a 556-bp intervening sequence (NTF-1). The amplification generates two amplicons from strain W, which indicate the presence and orientation of the NTF-1 sequence between the direct repeat of IS6110, and a third amplicon, which serves as an internal PCR control. The assay was evaluated with 193 isolates of M. tuberculosis, and all 48 strain W isolates among those 193 isolates were correctly identified.
authors.affiliation: Division of Bacterial and Mycotic Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333.
authors.forename: B B
authors.initials: BB
authors.lastname: Plikaytis
authors.forename: J L
authors.initials: JL
authors.lastname: Marden
authors.forename: J T
authors.initials: JT
authors.lastname: Crawford
authors.forename: C L
authors.initials: CL
authors.lastname: Woodley
authors.forename: W R
authors.initials: WR
authors.lastname: Butler
authors.forename: T M
authors.initials: TM
authors.lastname: Shinnick
mesh_terms: D000995:Antitubercular Agents / Q000494:pharmacology*; D001483:Base Sequence; D003428:Cross Infection / Q000453:epidemiology; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000032:analysis; D004196:Disease Outbreaks; D004352:Drug Resistance, Microbial; D005431:Florida / Q000453:epidemiology; D006801:Humans; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification*; D009518:New York / Q000453:epidemiology; D009519:New York City / Q000453:epidemiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D011330:Prisons; D012189:Retrospective Studies; D018088:Tuberculosis, Multidrug-Resistant / Q000188:drug therapy / Q000453:epidemiology / Q000382:microbiology*
chemical_list: D000995:Antitubercular Agents; D004251:DNA Transposable Elements; D004269:DNA, Bacterial