title: A consensus on fungal polymerase chain reaction diagnosis?: a United Kingdom-Ireland evaluation of polymerase chain reaction methods for detection of systemic fungal infections.
abstract: The limitations of classical diagnostic methods for invasive fungal infections (IFIs) have led to the development of molecular techniques to aid in the detection of IFIs. Despite good published performance, interlaboratory reproduction of these assays is variable, and no consensus has been reached for an optimal method. This publication describes the first multicenter study of polymerase chain reaction methods, for the detection of Aspergillus and Candida species, currently used in the UK and Ireland by distribution and analysis of multiple specimen control panels. All three Candida methods were comparable, achieving a satisfactory level of detection (10 cfu), and the method of preference was dependent on the requirements of the particular laboratory. The results for the five Aspergillus assays were more variable, but two methods (2Asp and 4Asp) were superior (10(1) conidia). Formally, the overall performances of the two Aspergillus assays were comparable (kappa statistic = 0.77). However, on the Roche LightCycler, there was a clear sample-type effect that greatly reduced the detection limit of the 4Asp method when testing whole blood samples. Therefore, the preferred Aspergillus method relied on the amplification platform available to the user. This study represents the initial process to achieve a consensus method for the diagnosis of IFIs.
authors.affiliation: Department of Medical Microbiology and NPHS Cardiff, University Hospital of Wales, Heath Park, Cardiff CF14 4XN, UK. lewis.white@nphs.wales.nhs.uk
authors.forename: P Lewis
authors.initials: PL
authors.lastname: White
authors.forename: Richard
authors.initials: R
authors.lastname: Barton
authors.forename: Malcolm
authors.initials: M
authors.lastname: Guiver
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Linton
authors.forename: Steve
authors.initials: S
authors.lastname: Wilson
authors.forename: Melvyn
authors.initials: M
authors.lastname: Smith
authors.forename: Beatriz L
authors.initials: BL
authors.lastname: Gomez
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Carr
authors.forename: Patrick T
authors.initials: PT
authors.lastname: Kimmitt
authors.forename: Shila
authors.initials: S
authors.lastname: Seaton
authors.forename: Kumar
authors.initials: K
authors.lastname: Rajakumar
authors.forename: Tessa
authors.initials: T
authors.lastname: Holyoake
authors.forename: Chris C
authors.initials: CC
authors.lastname: Kibbler
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Johnson
authors.forename: Richard P
authors.initials: RP
authors.lastname: Hobson
authors.forename: Brian
authors.initials: B
authors.lastname: Jones
authors.forename: Rosemary A
authors.initials: RA
authors.lastname: Barnes
mesh_terms: D001228:Aspergillosis / Q000175:diagnosis; D001483:Base Sequence; D002177:Candidiasis / Q000175:diagnosis; D032921:Consensus; D004271:DNA, Fungal / Q000032:analysis*; D006801:Humans; D007494:Ireland; D025202:Molecular Diagnostic Techniques / Q000295:instrumentation / Q000379:methods*; D008969:Molecular Sequence Data; D009181:Mycoses / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D012680:Sensitivity and Specificity; D012689:Sequence Homology, Nucleic Acid; D006113:United Kingdom
chemical_list: D004271:DNA, Fungal