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


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Title: Rapid and accurate detection of Pseudomonas aeruginosa by real-time polymerase chain reaction with melting curve analysis targeting gyrB gene.
Authors: Motoshima, Maiko / Yanagihara, Katsunori / Fukushima, Kazuko / Matsuda, Junichi / Sugahara, Kazuyuki / Hirakata, Yochi / Yamada, Yasuaki / Kohno, Shigeru / Kamihira, Shimeru
Issue Date: May-2007
Publisher: Elsevier Inc.
Citation: Diagnostic microbiology and infectious disease, 58(1), pp.53-58; 2007
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.
Keywords: Gyrase B / Melting temperature / Pseudomonas / Taxonomy
URI: http://hdl.handle.net/10069/22526
ISSN: 07328893
DOI: 10.1016/j.diagmicrobio.2006.11.007
PubMed ID: 17368797
Rights: Copyright © 2007 Elsevier Inc. All rights reserved.
Type: Journal Article
Text Version: author
Appears in Collections:Articles in academic journal

Citable URI : http://hdl.handle.net/10069/22526

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