title: Diagnostic value of a semi-nested PCR for the diagnosis of mucormycosis and aspergillosis from paraffin-embedded tissue: A single center experience.
abstract: OBJECTIVE
  The main aim of our study was to investigate the diagnostic value of a molecular method for the diagnosis of mucormycosis and aspergillosis from formalin-fixed and paraffin-embedded (FFPE) tissues.
  
  
  METHODS
  A retrospective chart review identified all cases with histology reports mentioning the presence of fungi with morphological characteristics of either Aspergillus or mucormycetes, for the period 2005-2012. Paraffin blocks were retrieved from the archives of the Department of Pathology. A semi-nested PCR specific for the detection of mucormycetes and Aspergillus species was applied in FFPE tissue from the above blocks. Results were compared with those of histological (gold standard) and microbiological methods.
  
  
  RESULTS
  Twenty cases with fungal hyphae in tissue were revealed. Mucormycetes were detected in 9 cases (45%) by PCR, in only 4 of which culture was available. Species of Aspergillus were detected in 8 cases (40%) by PCR, two of which were co-infection with mucormycetes. Five patients had other fungi, non-detectable with this specific PCR. At least one sample per patient was positive by PCR. Seven out of 30 samples tested overall were false negative. The calculated sensitivity of this method in our setting was 79.3% (95% CI: 60.3-91.9%); specificity was 100%.
  
  
  CONCLUSIONS
  The specific PCR used appears to be an easy and useful tool for the prompt and accurate diagnosis of mucormycosis and aspergillosis, in combination with histology and direct examination. Mucormycosis was more frequent than aspergillosis during the study period, highlighting the importance of continuous epidemiological surveillance of these serious infections.
authors.affiliation: Infectious Diseases Research Laboratory, 4th Dept. of Internal Medicine, National and Kapodistrian University of Athens, Medical School, "Attikon" University Hospital, Athens, Greece. Electronic address: mdrogari@hotmail.com.
authors.forename: Maria
authors.initials: M
authors.lastname: Drogari-Apiranthitou
authors.affiliation: 2nd Department of Pathology, National and Kapodistrian University of Athens, Medical School, "Attikon" University Hospital, Athens, Greece.
authors.forename: Ioannis
authors.initials: I
authors.lastname: Panayiotides
authors.affiliation: Infectious Diseases Research Laboratory, 4th Dept. of Internal Medicine, National and Kapodistrian University of Athens, Medical School, "Attikon" University Hospital, Athens, Greece.
authors.forename: Irene
authors.initials: I
authors.lastname: Galani
authors.affiliation: 2nd Department of Pathology, National and Kapodistrian University of Athens, Medical School, "Attikon" University Hospital, Athens, Greece.
authors.forename: Stefanos
authors.initials: S
authors.lastname: Konstantoudakis
authors.affiliation: Infectious Diseases Research Laboratory, 4th Dept. of Internal Medicine, National and Kapodistrian University of Athens, Medical School, "Attikon" University Hospital, Athens, Greece.
authors.forename: Georgios
authors.initials: G
authors.lastname: Arvanitidis
authors.affiliation: 2nd Department of Pathology, National and Kapodistrian University of Athens, Medical School, "Attikon" University Hospital, Athens, Greece.
authors.forename: Aris
authors.initials: A
authors.lastname: Spathis
authors.affiliation: 2nd Department of Pathology, National and Kapodistrian University of Athens, Medical School, "Attikon" University Hospital, Athens, Greece.
authors.forename: Alina-Roxani
authors.initials: AR
authors.lastname: Gouloumi
authors.affiliation: 2nd Department of Pathology, National and Kapodistrian University of Athens, Medical School, "Attikon" University Hospital, Athens, Greece.
authors.forename: Zoi
authors.initials: Z
authors.lastname: Tsakiraki
authors.affiliation: Infectious Diseases Research Laboratory, 4th Dept. of Internal Medicine, National and Kapodistrian University of Athens, Medical School, "Attikon" University Hospital, Athens, Greece.
authors.forename: Sotirios
authors.initials: S
authors.lastname: Tsiodras
authors.affiliation: Infectious Diseases Research Laboratory, 4th Dept. of Internal Medicine, National and Kapodistrian University of Athens, Medical School, "Attikon" University Hospital, Athens, Greece.
authors.forename: George
authors.initials: G
authors.lastname: Petrikkos
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001228:Aspergillosis / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009091:Mucormycosis / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods; D012189:Retrospective Studies; D012680:Sensitivity and Specificity
keywords: Aspergillosis; Invasive fungal infection; Mucormycosis; PCR; Paraffin embedded tissue