title: Comparison of three molecular assays for rapid detection of rifampin resistance in Mycobacterium tuberculosis.
abstract: Multidrug-resistant Mycobacterium tuberculosis (MDR-TB) is an emerging problem of great importance to public health, with higher mortality rates than drug-sensitive TB, particularly in immunocompromised patients. MDR-TB patients require treatment with more-toxic second-line drugs and remain infectious for longer than patients infected with drug-sensitive strains, incurring higher costs due to prolonged hospitalization. It is estimated that 90% of United Kingdom rifampin-resistant isolates are also resistant to isoniazid, making rifampin resistance a useful surrogate marker for multidrug resistance and indicating that second- and third-line drugs to which these isolates are susceptible are urgently required. Resistance in approximately 95% of rifampin-resistant isolates is due to mutations in a 69-bp region of the rpoB gene, making this a good target for molecular genotypic diagnostic methods. Two molecular assays, INNO-LiPA Rif.TB (Innogenetics, Zwijndrecht, Belgium) and MisMatch Detect II (Ambion, Austin, Tex.), were performed on primary specimens and cultures to predict rifampin resistance, and these methods were compared with the resistance ratio method. A third method, the phenotypic PhaB assay, was also evaluated in comparison to cultures in parallel with the genotypic assays. In an initial evaluation 16 of 16, 15 of 16, and 16 of 16 rifampin-resistant cultures (100, 93.8, and 100%, respectively), were correctly identified by line probe assay (LiPA), mismatch assay, and PhaB assay, respectively. Subsequently 38 sputa and bronchealveolar lavage specimens and 21 isolates were received from clinicians for molecular analysis. For the 38 primary specimens the LiPA and mismatch assay correlated with culture and subsequent identification and susceptibility tests in 36 and 38 specimens (94.7 and 100%), respectively. For the 21 isolates submitted by clinicians, both assays correlated 100% with routine testing.
authors.affiliation: Public Health Laboratory Service Mycobacterium Reference Unit, Department of Microbiology, King's College School of Medicine and Dentistry, King's College Hospital (Dulwich), London, United Kingdom. simon.watterson@kcl.ac.uk
authors.forename: S A
authors.initials: SA
authors.lastname: Watterson
authors.forename: S M
authors.initials: SM
authors.lastname: Wilson
authors.forename: M D
authors.initials: MD
authors.lastname: Yates
authors.forename: F A
authors.initials: FA
authors.lastname: Drobniewski
mesh_terms: D000904:Antibiotics, Antitubercular / Q000494:pharmacology*; D015373:Bacterial Typing Techniques; D001483:Base Sequence; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology; D004269:DNA, Bacterial / Q000032:analysis; D004352:Drug Resistance, Microbial / Q000235:genetics; D018432:Drug Resistance, Multiple / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D009160:Mycobacteriophages / Q000502:physiology; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000302:isolation & purification / Q000821:virology; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods; D012329:RNA, Bacterial / Q000032:analysis; D011933:Reagent Kits, Diagnostic; D012260:Ribonucleases; D012293:Rifampin / Q000494:pharmacology*; D017422:Sequence Analysis, DNA; D013183:Sputum / Q000382:microbiology; D018088:Tuberculosis, Multidrug-Resistant / Q000175:diagnosis / Q000382:microbiology*; D014397:Tuberculosis, Pulmonary / Q000175:diagnosis / Q000382:microbiology*
chemical_list: D000904:Antibiotics, Antitubercular; D004269:DNA, Bacterial; D012329:RNA, Bacterial; D011933:Reagent Kits, Diagnostic; D012260:Ribonucleases; D012293:Rifampin