title: Development of an immunochromatographic assay for the rapid detection of AAC(6')-Iae-producing multidrug-resistant Pseudomonas aeruginosa.
abstract: OBJECTIVE
  To develop an easy-to-use method for the rapid detection of antibiotic-resistant bacteria. Here, a new immunochromatographic assay specific for aminoglycoside 6'-N-acetyltransferase AAC(6')-Iae was designed. AAC(6')-Iae is a significant marker molecule for multidrug-resistant (MDR) Pseudomonas aeruginosa isolates in Japan.
  
  
  METHODS
  Monoclonal antibodies specific for AAC(6')-Iae were used to construct the assay. The assessment of the assay was performed using 116 P. aeruginosa clinical isolates obtained from hospitals in the Kanto area of Japan where little was known about AAC(6')-Iae producers. PCR analyses of the aac(6')-Iae and class 1 integron, antimicrobial susceptibility testing and PFGE analysis were performed to characterize positive strains.
  
  
  RESULTS
  The detection limit of the assay was 1.0 x 10(5) cfu. Of 116 clinical isolates, 60 were positive for AAC(6')-Iae using the assay. The results of assessment with clinical isolates were fully consistent with those of aac(6')-Iae PCR analyses, showing no false positives or negatives. All positive strains detected by the assay showed MDR phenotypes that were resistant to several classes of antibiotic. PFGE analysis showed that 59 of 60 positive strains tightly clustered, and these included clonal expansions.
  
  
  CONCLUSIONS
  The developed assay is an easy-to-use and reliable detection method for AAC(6')-Iae-producing MDR P. aeruginosa. This approach may be applicable for screening and investigation of antibiotic-resistant bacteria as an alternative to PCR analysis.
authors.affiliation: Department of Infectious Diseases, National Center for Global Health and Medicine, Shinjuku, Tokyo 162-8655, Japan.
authors.forename: Tomoe
authors.initials: T
authors.lastname: Kitao
authors.forename: Tohru
authors.initials: T
authors.lastname: Miyoshi-Akiyama
authors.forename: Kayo
authors.initials: K
authors.lastname: Shimada
authors.forename: Masashi
authors.initials: M
authors.lastname: Tanaka
authors.forename: Kenji
authors.initials: K
authors.lastname: Narahara
authors.forename: Nobuko
authors.initials: N
authors.lastname: Saito
authors.forename: Teruo
authors.initials: T
authors.lastname: Kirikae
mesh_terms: D000123:Acetyltransferases / Q000032:analysis*; D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins / Q000032:analysis; D015373:Bacterial Typing Techniques; D001431:Bacteriological Techniques / Q000379:methods*; D002845:Chromatography / Q000379:methods*; D016000:Cluster Analysis; D003428:Cross Infection / Q000382:microbiology; D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000235:genetics; D024901:Drug Resistance, Multiple, Bacterial; D016521:Electrophoresis, Gel, Pulsed-Field; D005838:Genotype; D006801:Humans; D007118:Immunoassay / Q000379:methods; D032023:Integrons; D007564:Japan; D016133:Polymerase Chain Reaction; D011552:Pseudomonas Infections / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000302:isolation & purification; D012680:Sensitivity and Specificity
chemical_list: D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D000123:Acetyltransferases; C056532:aminoglycoside N(6')-acetyltransferase