title: Development of an immunochromatographic assay for diagnosing the production of IMP-type metallo-β-lactamases that mediate carbapenem resistance in Pseudomonas.
abstract: Rapid and reliable detection of carbapenem-resistant bacteria is an important infection-control measure and a crucial aspect of antimicrobial chemotherapy. IMP-type metallo-β-lactamase (MBL) is an emzyme that mediate carbapenem resistance in bacteria. Here, an immunochromatographic assay was newly developed using novel monoclonal antibodies (mAbs) recognizing IMP-type MBL. Epitope mapping of mAbs and mutational analysis of the epitope region in IMP antigen suggested that the mAbs could react to all known subtypes of IMP-type MBL. Evaluation of the assay using Pseudomonas aeruginosa strains (n=248) showed that the results of the immunochromatographic detection of the IMP-type MBLs were fully consistent with those of the PCR analysis for bla(IMP) genes, showing false positives and negatives. All positive strains were resistant to carbapenem (MIC ≥ 16 μg/ml). The assay also accurately distinguished the production of IMP-type MBLs in Pseudomonas putida, Acinetobacter baumannii, and Alcaligenes xylosoxidans. The detection limit of the assay was 5.7×10(4)cfu per test. Taken together, these data suggest that the developed assay can be used for rapid and reliable diagnosis of the production of IMP-type MBLs in Gram-negative bacteria.
authors: Department of Infectious Diseases, Research Institute, National Center for Global Health and Medicine, Shinjuku, Tokyo, Japan.
authors: Tomoe
authors: T
authors: Kitao
authors: Tohru
authors: T
authors: Miyoshi-Akiyama
authors: Masashi
authors: M
authors: Tanaka
authors: Kenji
authors: K
authors: Narahara
authors: Masahiro
authors: M
authors: Shimojima
authors: Teruo
authors: T
authors: Kirikae
mesh_terms: D040981:Acinetobacter baumannii / Q000201:enzymology; D000421:Alcaligenes / Q000201:enzymology; D000911:Antibodies, Monoclonal; D015780:Carbapenems / Q000494:pharmacology*; D002846:Chromatography, Affinity / Q000379:methods*; D018604:Epitope Mapping; D000939:Epitopes / Q000235:genetics / Q000276:immunology; D008826:Microbial Sensitivity Tests; D020125:Mutation, Missense; D011549:Pseudomonas / Q000201:enzymology*; D018440:beta-Lactam Resistance; D001618:beta-Lactamases / Q000032:analysis*
chemical_list: D000911:Antibodies, Monoclonal; D015780:Carbapenems; D000939:Epitopes; D001618:beta-Lactamases