title: [Determination of genetic mutations in rpoB of the isolates of Mycobacterium tuberculosis complex those susceptibilities to rifampicin were interpreted as indeterminate by BrothMIC MTB].
abstract: The minimum inhibitory concentrations (MICs) to rifampicin (RFP) for Mycobacterium tuberculosis complex distribute bipartitely, the most susceptible, wild isolates showing < or = 0.03 microg/ml of MICs and the resistant isolates are >2.0 microg/ml. As the results, a very few number of isolates showing the MICs between 0.06 microg/ml to 2.0 microg/ml are still interpreted as "indeterminate" by BrothMIC MTB. In this communication, we determined genetic mutations in rpoB gene of the isolates of M. tuberculosis complex those MICs to RFP resulted in "indeterminate" interpretations. Through the direct base-sequencing, genetic mutation (s) in rpoB gene associated with amino acid substitution(s) were found in 21 of 27 clinical isolates of M. tuberculosis complex. All the isolates with 0.06 microg/ml of MICs were confirmed as being a wild type, whereas those with > or =0.125 microg/ml of MICs have a variety of genetic mutations. There found one exception, that is, a strain with 0.5 microg/ml of MIC revealed no mutation in rpoB gene. In addition to direct base-sequencing, a line probe assay, LiPA . Rif TB (Innogenetics N.V., Zwijnaarde, Belgium) was comparatively evaluated. The results obtained were highly correlated, LiPA . Rif TB giving comparable readings for 26 (96.3S) of 27 isolates tested. With these results, it can be concluded that the interpretive breakpoints for RFP MICs determined by BrothMIC MTB should be revised as follows: susceptible, < or =0.06 microg/ml, and resistant, > or =0.125 microg/ml. Secondly, LiPA . Rif TB gave an accurate and rapid interpretation for RFP resistance, but it may be concerned for the occasional false-susceptible readings.
authors.affiliation: Department of Laboratory Medicine, Graduate School and Faculty of Medicine, University of the Ryukyus, 207 Uehara, Nishihara, Nakagami-Okinawa 903-0215, Japan. chika@med.u-ryukyu.ac.jp
authors.forename: Chika M
authors.initials: CM
authors.lastname: Shiohira
authors.forename: Nobuhisa
authors.initials: N
authors.lastname: Yamane
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D012321:DNA-Directed RNA Polymerases; D024881:Drug Resistance, Bacterial / Q000235:genetics; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics*; D012293:Rifampin / Q000494:pharmacology*