title: Detection of Aspergillus flavus and A. fumigatus in Bronchoalveolar Lavage Specimens of Hematopoietic Stem Cell Transplants and Hematological Malignancies Patients by Real-Time Polymerase Chain Reaction, Nested PCR and Mycological Assays.
abstract: BACKGROUND
  Pulmonary aspergillosis (PA) is one of the most serious complications in immunocompromised patients, in particular among hematopoietic stem cell transplants (HSCT) and patients with hematological malignancies.
  
  
  OBJECTIVE
  The current study aimed to evaluate the incidence of PA and utility of molecular methods in HSCT and patients with hematological malignancies, four methods including direct examination, culture, nested polymerase chain reaction (PCR) and real-time PCR were performed on bronchoalveolar lavage (BAL) specimens in Tehran, Iran.
  
  
  METHODS
  During 16 months, 46 BAL specimens were obtained from individuals with allogeneic HSCT (n = 18) and patients with hematological malignancies (n = 28). Direct wet mounts with 20% potassium hydroxide (KOH) and culture on mycological media were performed. The molecular detection of Aspergillus fumigatus and A. flavus was done by amplifying the conserved sequences of internal transcribed spacer 1 (ITS1) ribosomal DNA by nested-PCR and the β-tubulin gene by TaqMan real-time PCR.
  
  
  RESULTS
  Seven (15.2%) out of 46 specimens were positive in direct examination and showed branched septate hyphae; 11 (23.9%) had positive culture including eight (72.7%) A. flavus and three (27.3%) A. fumigatus; 22 (47.8%) had positive nested-PCR and eight (17.4%) had positive real-time PCR. The incidence of invasive pulmonary aspergillosis (IPA) in these patients included proven IPA in 1 (2.2%), probable IPA in 10 (21.7%), possible IPA in 19 (41.3%) and not IPA in 16 cases (34.8%).
  
  
  CONCLUSIONS
  The incidence of IPA in allogeneic HSCT and patients with hematological malignancies was relatively high and A. flavus was the most common cause of PA. As molecular methods had higher sensitivity, it may be useful as screening methods in HSCT and patients with hematological malignancies, or to determine when empirical antifungal therapy can be withheld.
authors.affiliation: Allergy Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, IR Iran ; Department of Parasitology and Mycology, Ghaem Hospital, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, IR Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Zarrinfar
authors.affiliation: Department of Medical Parasitology and Mycology, Tehran University of Medical Sciences, Tehran, IR Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Mirhendi
authors.affiliation: Research Center for Skin Diseases and Cutaneous Leishmaniasis, Department of Parasitology and Mycology, Mashhad University of Medical Sciences, Mashhad, IR Iran.
authors.forename: Abdolmajid
authors.initials: A
authors.lastname: Fata
authors.affiliation: Department of Parasitology and Mycology, Shiraz University of Medical Sciences, Shiraz, IR Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Khodadadi
authors.affiliation: Department of Medical Parasitology and Mycology, Tehran University of Medical Sciences, Tehran, IR Iran.
authors.forename: Parivash
authors.initials: P
authors.lastname: Kordbacheh
keywords: Aspergillus flavus; Aspergillus fumigatus; Bronchoalveolar Lavage; Hematological Malignancies; Hematopoietic Stem Cell Transplantation; Invasive Pulmonary Aspergillosis