title: 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 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.
authors: Department of Laboratory Medicine, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki City, Nagasaki 852-8501, Japan.
authors: Maiko
authors: M
authors: Motoshima
authors: Katsunori
authors: K
authors: Yanagihara
authors: Kazuko
authors: K
authors: Fukushima
authors: Junichi
authors: J
authors: Matsuda
authors: Kazuyuki
authors: K
authors: Sugahara
authors: Yochi
authors: Y
authors: Hirakata
authors: Yasuaki
authors: Y
authors: Yamada
authors: Shigeru
authors: S
authors: Kohno
authors: Shimeru
authors: S
authors: Kamihira
mesh_terms: D015373:Bacterial Typing Techniques; D001483:Base Sequence; D027081:DNA Gyrase / Q000235:genetics*; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011552:Pseudomonas Infections / Q000175:diagnosis* / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013997:Time Factors; D044366:Transition Temperature
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D027081:DNA Gyrase