title: Clinical evaluation of a Mucorales-specific real-time PCR assay in tissue and serum samples.
abstract: Molecular diagnostic assays can accelerate the diagnosis of fungal infections and subsequently improve patient outcomes. In particular, the detection of infections due to Mucorales is still challenging for laboratories and physicians. The aim of this study was to evaluate a probe-based Mucorales-specific real-time PCR assay (Muc18S) using tissue and serum samples from patients suffering from invasive mucormycosis (IMM). This assay can detect a broad range of clinically relevant Mucorales species and can be used to complement existing diagnostic tests or to screen high-risk patients. An advantage of the Muc18S assay is that it exclusively detects Mucorales species allowing the diagnosis of Mucorales DNA without sequencing within a few hours. In paraffin-embedded tissue samples this PCR-based method allowed rapid identification of Mucorales in comparison with standard methods and showed 91 % sensitivity in the IMM tissue samples. We also evaluated serum samples, an easily accessible material, from patients at risk from IMM. Mucorales DNA was detected in all patients with probable/proven IMM (100 %) and in 29 % of the possible cases. Detection of IMM in serum could enable an earlier diagnosis (up to 21 days) than current methods including tissue samples, which were gained mainly post-mortem. A screening strategy for high-risk patients, which would enable targeted treatment to improve patient outcomes, is therefore possible.
authors.affiliation: Department for Internal Medicine II, University Hospital of Wuerzburg, Wuerzburg, Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: Springer
authors.affiliation: Division of Hygiene and Medical Microbiology, Medical University of Innsbruck, Austria.
authors.forename: Michaela
authors.initials: M
authors.lastname: Lackner
authors.affiliation: Division of Hygiene and Medical Microbiology, Medical University of Innsbruck, Austria.
authors.forename: Christian
authors.initials: C
authors.lastname: Ensinger
authors.affiliation: Division of Hygiene and Medical Microbiology, Medical University of Innsbruck, Austria.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Risslegger
authors.affiliation: School of Science and Health, Western Sydney University, Australia.
authors.forename: Charles Oliver
authors.initials: CO
authors.lastname: Morton
authors.affiliation: University Hospital of Internal Medicine V, Hematology & Oncology, Medical University of Innsbruck, Austria.
authors.forename: David
authors.initials: D
authors.lastname: Nachbaur
authors.affiliation: Division of Hygiene and Medical Microbiology, Medical University of Innsbruck, Austria.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Lass-Flörl
authors.affiliation: Department for Internal Medicine II, University Hospital of Wuerzburg, Wuerzburg, Germany.
authors.forename: Hermann
authors.initials: H
authors.lastname: Einsele
authors.affiliation: Department for Internal Medicine II, University Hospital of Wuerzburg, Wuerzburg, Germany.
authors.forename: Werner J
authors.initials: WJ
authors.lastname: Heinz
authors.affiliation: Department for Internal Medicine II, University Hospital of Wuerzburg, Wuerzburg, Germany.
authors.forename: Juergen
authors.initials: J
authors.lastname: Loeffler
mesh_terms: D000328:Adult; D000368:Aged; D015342:DNA Probes; D004271:DNA, Fungal / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D005260:Female; D016469:Fungemia / Q000382:microbiology; D006801:Humans; D000072742:Invasive Fungal Infections / Q000175:diagnosis / Q000382:microbiology; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009090:Mucorales / Q000235:genetics / Q000302:isolation & purification*; D009091:Mucormycosis / Q000175:diagnosis* / Q000382:microbiology; D016612:Paraffin Embedding; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012307:Risk Factors; D055815:Young Adult
chemical_list: D015342:DNA Probes; D004271:DNA, Fungal; D012337:RNA, Ribosomal, 18S