title: Evaluation of repetitive sequence PCR and PCR-mass spectrometry for the identification of clinically relevant Candida species.
abstract: The application of molecular diagnostic methods may improve the timeliness and accuracy with which fungi are identified. A total of 76 well-characterized reference strains of clinically relevant Candida species and 61 clinical Candida isolates were tested by repetitive sequence PCR (rep-PCR) and PCR followed by electrospray ionization mass spectrometry (PCR/ESI-MS) and results compared against internal transcribed spacer (ITS) ribosomal RNA gene sequencing as a reference standard. Both rep-PCR and PCR/ESI-MS correctly identified 51 isolates to the species level. When method performance was evaluated based only on genospecies included in the reference libraries, both methods yielded an accuracy of 98.1%. It may be concluded that rep-PCR and PCR/ESI-MS are highly effective at identifying clinical isolates of Candida to the species level. These methods hold promise for improving the speed and accuracy of identification of Candida spp. in clinical mycology laboratories.
authors.affiliation: Department of Pathology, St. Jude Children's Research Hospital , Memphis, TN, USA.
authors.forename: Zhengming
authors.initials: Z
authors.lastname: Gu
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Hall
authors.forename: Mark
authors.initials: M
authors.lastname: Frinder
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Walsh
authors.forename: Randall T
authors.initials: RT
authors.lastname: Hayden
mesh_terms: D002175:Candida / Q000737:chemistry / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D002177:Candidiasis / Q000175:diagnosis* / Q000382:microbiology; D006801:Humans; D008828:Microbiological Techniques / Q000379:methods*; D009172:Mycology / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*