Development of TaqMan real-time polymerase chain reaction for the detection and identification of Penicillium marneffei.
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.
Pornprasert
Sakorn
S
Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang-Mai University, Chiang-Mai, Thailand. sakornmi001@yahoo.com
Praparattanapan
Jutarat
J
Khamwan
Chantana
C
Pawichai
Sudjai
S
Pimsarn
Parichat
P
Samleerat
Tanawan
T
Leechanachai
Pranee
P
Supparatpinyo
Khunchai
K
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
D017931:DNA Primers; D004271:DNA, Fungal; D004275:DNA, Ribosomal; D012340:RNA, Ribosomal, 5.8S