Development of an immunochromatographic assay for the rapid detection of AAC(6′)-Iae-producing multidrug-resistant Pseudomonas aeruginosa

Autor: Tohru Miyoshi-Akiyama, Teruo Kirikae, Kenji Narahara, Masashi Tanaka, Nobuko Saito, Kayo Shimada, Tomoe Kitao
Rok vydání: 2010
Předmět:
DNA
Bacterial
Microbiology (medical)
Genotype
Drug resistance
medicine.disease_cause
Integron
Polymerase Chain Reaction
Sensitivity and Specificity
Integrons
law.invention
Microbiology
Bacterial Proteins
Japan
Acetyltransferases
law
Drug Resistance
Multiple
Bacterial
medicine
Cluster Analysis
Humans
Pseudomonas Infections
Pharmacology (medical)
Polymerase chain reaction
Immunoassay
Pharmacology
Bacteriological Techniques
Chromatography
Cross Infection
biology
Pseudomonas aeruginosa
Antibodies
Monoclonal
biochemical phenomena
metabolism
and nutrition
bacterial infections and mycoses
biology.organism_classification
Antibodies
Bacterial
DNA Fingerprinting
Bacterial Typing Techniques
Electrophoresis
Gel
Pulsed-Field
Multiple drug resistance
Infectious Diseases
DNA profiling
biology.protein
Pseudomonadaceae
Zdroj: Journal of Antimicrobial Chemotherapy. 65:1382-1386
ISSN: 1460-2091
0305-7453
Popis: Objectives 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.
Databáze: OpenAIRE
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