title: Detection of Candida and Aspergillus species DNA using broad-range real-time PCR for fungal endophthalmitis.
abstract: BACKGROUND
  The goal of this work is to establish a broad-range real-time polymerase chain reaction (PCR) diagnostic system for ocular fungal infection and to measure Candida and Aspergillus DNA in the ocular fluids obtained from unknown uveitis/endophthalmitis patients.
  
  
  METHODS
  After obtaining informed consent, intraocular fluids (aqueous humor and vitreous fluid samples) were collected from 54 patients with idiopathic uveitis or endophthalmitis. Samples were assayed for Candida or Aspergillus DNA using broad-range (18S rRNA sequences) quantitative real-time PCR.
  
  
  RESULTS
  Candida or Aspergillus DNA was detected in seven out of 54 patient ocular samples (13%). These PCR-positive samples showed significantly high copy numbers of Candida or Aspergillus DNA. On the other hand, fungal DNA was not detected in any of the other 46 samples collected from these idiopathic uveitis or endophthalmitis patients. In the one PCR-negative case, PCR did not detect any fungal genome in the sample, even though this patient was clinically suspected of having Candida endophthalmitis. Real-time PCR results were negative for fungal DNA in the bacterial endophthalmitis patients and in various uveitis patients. In addition, fungal DNA was also not detected in patients without ocular inflammation (controls).
  
  
  CONCLUSIONS
  Analysis of ocular samples by this broad-range real-time PCR method can be utilized for rapid diagnosis of patients suffering from unknown intraocular disorders such as idiopathic uveitis/endophthalmitis.
authors.affiliation: Department of Ophthalmology & Visual Science, Tokyo Medical and Dental University, Graduate School of Medical and Dental Sciences, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan. sunaoph@tmd.ac.jp
authors.forename: Sunao
authors.initials: S
authors.lastname: Sugita
authors.forename: Koju
authors.initials: K
authors.lastname: Kamoi
authors.forename: Manabu
authors.initials: M
authors.lastname: Ogawa
authors.forename: Ken
authors.initials: K
authors.lastname: Watanabe
authors.forename: Norio
authors.initials: N
authors.lastname: Shimizu
authors.forename: Manabu
authors.initials: M
authors.lastname: Mochizuki
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000935:Antifungal Agents / Q000627:therapeutic use; D001082:Aqueous Humor / Q000382:microbiology; D001228:Aspergillosis / Q000175:diagnosis / Q000188:drug therapy / Q000382:microbiology*; D001230:Aspergillus / Q000235:genetics* / Q000302:isolation & purification; D002175:Candida / Q000235:genetics* / Q000302:isolation & purification; D002177:Candidiasis / Q000175:diagnosis / Q000188:drug therapy / Q000382:microbiology*; D004271:DNA, Fungal / Q000032:analysis*; D009877:Endophthalmitis / Q000175:diagnosis / Q000188:drug therapy / Q000382:microbiology*; D015821:Eye Infections, Fungal / Q000175:diagnosis / Q000188:drug therapy / Q000382:microbiology*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014822:Vitreous Body / Q000382:microbiology
chemical_list: D000935:Antifungal Agents; D004271:DNA, Fungal; D012337:RNA, Ribosomal, 18S