title: Identification of Azole Resistance Markers in Clinical Isolates of Candida tropicalis Using cDNA-AFLP Method.
abstract: BACKGROUND
  Global reports have highlighted the increasing prevalence of Candida tropicalis infections as well as organism(') s drug resistance. This study aimed at identifying azole resistance markers in clinical isolates of C. tropicalis, which will be a great resource for developing new drugs.
  
  
  METHODS
  Two susceptible and resistant isolates of C. tropicalis were recovered from an epidemiological investigation of candidiasis in immunocompromised patients. C. tropicalis ATCC 750 was used as reference strain. Antifungal susceptibility to fluconazole and itraconazole was determined using Clinical and Laboratory Standards Institute (CLSI) method. Complementary DNA-amplified fragment length polymorphism (cDNA-AFLP) technology and real-time reverse-transcriptase (RT) PCR were used for identification of potential genes involved in azole resistance of C. tropicalis clinical isolates.
  
  
  RESULTS
  Five genes encoding the following enzymes were identified as superoxide dismutase (SOD) implicated in antioxidant defense, ornithine aminotransferase (OAT), acetyl ornithine aminotransferase (ACOAT), adenosylmethionine-8-amino-7-oxononanoate aminotransferase (DAPA AT), and 4-aminobutyrate aminotransferase (ABAT)-belonging to pyridoxal phosphate (PLP) dependent enzymes and acting in an important physiological role in many fungal-cell cycles. Real-time RT-PCR confirmed mRNA level of the aforementioned genes.
  
  
  CONCLUSIONS
  Our findings showed that factors such as PLP-dependent enzymes and SOD might be implicated in drug resistance in C. tropicalis clinical isolate. Therefore, further studies are required to explore the accurate biological functions of the mentioned genes that would be helpful for effective drug development.
authors: Department of Medical Mycology and Parasitology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors: Ali
authors: A
authors: Kanani
authors: Department of Medical Mycology and Parasitology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors: Farideh
authors: F
authors: Zaini
authors: Department of Medical Mycology and Parasitology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors: Parivash
authors: P
authors: Kordbacheh
authors: Department of Medical Mycology and Parasitology, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran.
authors: Mehraban
authors: M
authors: Falahati
authors: Department of Medical Mycology and Parasitology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors: Sassan
authors: S
authors: Rezaie
authors: Department of Medical Mycology and Parasitology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors: Roshanak
authors: R
authors: Daie
authors: Department of Medical Mycology and Parasitology, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran.
authors: Shirin
authors: S
authors: Farahyar
authors: Department of Medical Mycology and Parasitology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors: Mahin
authors: M
authors: Safara
authors: Department of Microbiology and Immunology, Faculty of Medicine, Qom University of Medical Sciences, Qom, Iran.
authors: Roohollah
authors: R
authors: Fateh
authors: Department of Virology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors: Ebrahim
authors: E
authors: Faghihloo
authors: Department of Medical Mycology and Parasitology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors: Azam
authors: A
authors: Fattahi
authors: Department of Medical Genetics, Tehran University of Medical Sciences, Tehran, Iran.
authors: Mansour
authors: M
authors: Heidari
mesh_terms: D054458:Amplified Fragment Length Polymorphism Analysis / Q000379:methods*; D001393:Azoles / Q000494:pharmacology*; D041022:Candida tropicalis / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification*; D018076:DNA, Complementary / Q000235:genetics*; D025141:Drug Resistance, Fungal / Q000187:drug effects*; D004591:Electrophoresis, Polyacrylamide Gel; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D005819:Genetic Markers; D006801:Humans; D008826:Microbial Sensitivity Tests
chemical_list: D001393:Azoles; D018076:DNA, Complementary; D005819:Genetic Markers
keywords: azole resistance; candidiasis; differential display; real-time RT-PCR