title: Evaluation of nested PCR in diagnosis of fungal rhinosinusitis.
abstract: OBJECTIVE
  Given the importance of rapid diagnosis for fungal rhinosinusitis, this study aimed to evaluate the use of nested PCR to identify Aspergillus and Mucor species in clinical samples from patients with suspected fungal rhinosinusitis.
  
  
  METHODS
  Functional endoscopic sinus surgery specimens were collected from 98 patients with rhinosinusitis from 2012 to 2013. All samples were ground and cultured on sabouraud dextrose agar. The isolated fungi were identified based on their macroscopic and microscopic features. Fungal DNA was extracted from the tissue samples and nested PCR was performed with two sets of primers for Mucor and Aspergillus.
  
  
  RESULTS
  Direct microscopic showed that 5.1% contained fungal components and 9.2% exhibited growth of fungi in culture. The most common agents isolated were Aspergillus fumigatus (n= 3), Aspergillus flavus (n=2), Penicillium sp (n=3) and Alternaria sp. (n=1). Mucor sp. was identified in the pathology smear from 1 patient. Positive results for fungal rhinosinusitis were obtained for a total of 10.2% by culture or pathology smear. Positive PCR results were obtained in 72 samples for Aspergillus and 31 samples for Mucor.
  
  
  CONCLUSIONS
  Our results suggest that endoscopic sinus surgery specimens are not suitable for nested PCR, probably because of the accumulation of fungi that contaminate the environmental air. This drawback is a limiting factor for diagnosis with nasal cavity specimens. Therefore, molecular methods and conventional culture techniques are helpful complementary diagnostic methods to detect fungal rhinosinusitis and determine appropriate management for these patients.
authors.affiliation: Professor Alborzi Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
authors.forename: Parisa
authors.initials: P
authors.lastname: Badiee
authors.affiliation: Otolaryngology Unit, Shiraz University of Medical Sciences, Shiraz, Iran.
authors.forename: Behrooz
authors.initials: B
authors.lastname: Gandomi
authors.affiliation: Otolaryngology Unit, Shiraz University of Medical Sciences, Shiraz, Iran.
authors.forename: Gholamabbass
authors.initials: G
authors.lastname: Sabz
authors.affiliation: Otolaryngology Unit, Shiraz University of Medical Sciences, Shiraz, Iran.
authors.forename: Bijan
authors.initials: B
authors.lastname: Khodami
authors.affiliation: Professor Alborzi Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
authors.forename: Maral
authors.initials: M
authors.lastname: Choopanizadeh
authors.affiliation: Professor Alborzi Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
authors.forename: Hadis
authors.initials: H
authors.lastname: Jafarian
keywords: Aspergillus; functional endoscopic sinus surgery; nested PCR; rhinosinusitis