Rapid and accurate detection of Pseudomonas aeruginosa by real-time polymerase chain reaction with melting curve analysis targeting gyrB gene

Abstract Laboratory detection of Pseudomonas spp., particularly Pseudomonas aeruginosa , is an important assay in the nosocomial control. The study was designed firstly to establish a new assay-applied LightCycler polymerase chain reaction (PCR) technology with melting curve analysis (MCA). A total...

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Bibliographic Details
Published in:Diagnostic microbiology and infectious disease 2007-05, Vol.58 (1), p.53-58
Main Authors: Motoshima, Maiko, Yanagihara, Katsunori, Fukushima, Kazuko, Matsuda, Junichi, Sugahara, Kazuyuki, Hirakata, Yochi, Yamada, Yasuaki, Kohno, Shigeru, Kamihira, Shimeru
Format: Article
Language:English
Subjects:
Bacterial Typing Techniques
Bacteriology
Base Sequence
Biological and medical sciences
DNA Gyrase - genetics
DNA Primers
DNA, Bacterial - analysis
Fundamental and applied biological sciences. Psychology
Gyrase B
Humans
Infectious Disease
Infectious diseases
Internal Medicine
Medical sciences
Melting temperature
Microbiology
Miscellaneous
Molecular Sequence Data
Polymerase Chain Reaction - methods
Pseudomonas
Pseudomonas aeruginosa
Pseudomonas aeruginosa - classification
Pseudomonas aeruginosa - genetics
Pseudomonas aeruginosa - isolation & purification
Pseudomonas Infections - diagnosis
Pseudomonas Infections - microbiology
Pseudomonas otitidis
Sensitivity and Specificity
Taxonomy
Time Factors
Transition Temperature
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Summary:Abstract Laboratory detection of Pseudomonas spp., particularly Pseudomonas aeruginosa , is an important assay in the nosocomial control. The study was designed firstly to establish a new assay-applied LightCycler polymerase chain reaction (PCR) technology with melting curve analysis (MCA). A total of 224 Gram-negative isolates were used to verify the assay system. The PCR with MCA method using the P. aeruginosa -specific gyrase B gene primers was rapid and accurate; the total run is approximately 3 h, and the sensitivity and specificity relative to the Vitek (bioMerieux, Hazelwood, MO) results were 98.1% and 100%, respectively. Vitek identification system was not able to identify the isolates from the new Pseudomonas otitidis spp. opposite to the real-time PCR. This assay was validated to be accurate with an overall sensitivity and specificity of 98.7% and 98.9%, respectively. Conclusively, this rapid and accurate PCR assay with MCA will help to manage and control infections with P. aeruginosa.
ISSN:0732-8893
1879-0070
DOI:10.1016/j.diagmicrobio.2006.11.007