title: Development of TaqMan real-time polymerase chain reaction for the detection and identification of Penicillium marneffei.
abstract: Penicillium marneffei is a dimorphic fungus, which is endemic in Southeast Asia and responsible for emerging opportunistic infections. Diagnosis of penicilliosis may be difficult when few yeast cells are present, while a gold standard diagnosis technique requires long-term culture. In order to provide a more rapid and accurate diagnosis, we developed a TaqMan real-time PCR to detect and identify P. marneffei DNA coding for 5.8S rRNA in purified yeast DNA and clinical samples. All P. marneffei DNA preparations could be detected using specific primers and TaqMan probe. The assay has a sensitivity to detect at least 10 yeast cells in seeded blood. Moreover, it can detect P. marneffei DNA in peripheral blood samples and blood-culture bottles. Therefore, the real-time PCR assay may represent a potential tool for early diagnosis of penicilliosis marneffei.
authors.affiliation: Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang-Mai University, Chiang-Mai, Thailand. sakornmi001@yahoo.com
authors.forename: Sakorn
authors.initials: S
authors.lastname: Pornprasert
authors.forename: Jutarat
authors.initials: J
authors.lastname: Praparattanapan
authors.forename: Chantana
authors.initials: C
authors.lastname: Khamwan
authors.forename: Sudjai
authors.initials: S
authors.lastname: Pawichai
authors.forename: Parichat
authors.initials: P
authors.lastname: Pimsarn
authors.forename: Tanawan
authors.initials: T
authors.lastname: Samleerat
authors.forename: Pranee
authors.initials: P
authors.lastname: Leechanachai
authors.forename: Khunchai
authors.initials: K
authors.lastname: Supparatpinyo
mesh_terms: D001210:Asia, Southeastern; D001769:Blood / Q000382:microbiology; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009172:Mycology / Q000379:methods*; D009181:Mycoses / Q000175:diagnosis* / Q000382:microbiology*; D010407:Penicillium / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012340:RNA, Ribosomal, 5.8S / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D004271:DNA, Fungal; D004275:DNA, Ribosomal; D012340:RNA, Ribosomal, 5.8S