title: PCR followed by electrospray ionization mass spectrometry for broad-range identification of fungal pathogens.
abstract: Invasive fungal infections are a significant cause of morbidity and mortality among immunocompromised patients. Early and accurate identification of these pathogens is central to direct therapy and to improve overall outcome. PCR coupled with electrospray ionization mass spectrometry (PCR/ESI-MS) was evaluated as a novel means for identification of fungal pathogens. Using a database grounded by 60 ATCC reference strains, a total of 394 clinical fungal isolates (264 molds and 130 yeasts) were analyzed by PCR/ESI-MS; results were compared to phenotypic identification, and discrepant results were sequence confirmed. PCR/ESI-MS identified 81.4% of molds to either the genus or species level, with concordance rates of 89.7% and 87.4%, respectively, to phenotypic identification. Likewise, PCR/ESI-MS was able to identify 98.4% of yeasts to either the genus or species level, agreeing with 100% of phenotypic results at both the genus and species level. PCR/ESI-MS performed best with Aspergillus and Candida isolates, generating species-level identification in 94.4% and 99.2% of isolates, respectively. PCR/ESI-MS is a promising new technology for broad-range detection and identification of medically important fungal pathogens that cause invasive mycoses.
authors.affiliation: Ibis Biosciences, Abbott Laboratories, Carlsbad, California, USA.
authors.forename: Christian
authors.initials: C
authors.lastname: Massire
authors.forename: Daelynn R
authors.initials: DR
authors.lastname: Buelow
authors.forename: Sean X
authors.initials: SX
authors.lastname: Zhang
authors.forename: Robert
authors.initials: R
authors.lastname: Lovari
authors.forename: Heather E
authors.initials: HE
authors.lastname: Matthews
authors.forename: Donna M
authors.initials: DM
authors.lastname: Toleno
authors.forename: Raymond R
authors.initials: RR
authors.lastname: Ranken
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Hall
authors.forename: David
authors.initials: D
authors.lastname: Metzgar
authors.forename: Rangarajan
authors.initials: R
authors.lastname: Sampath
authors.forename: Lawrence B
authors.initials: LB
authors.lastname: Blyn
authors.forename: David J
authors.initials: DJ
authors.lastname: Ecker
authors.forename: Zhengming
authors.initials: Z
authors.lastname: Gu
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Walsh
authors.forename: Randall T
authors.initials: RT
authors.lastname: Hayden
mesh_terms: D005658:Fungi / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008828:Microbiological Techniques / Q000379:methods*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009172:Mycology / Q000379:methods*; D009181:Mycoses / Q000175:diagnosis / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*