title: Diagnosis of ocular toxoplasmosis by two polymerase chain reaction (PCR) examinations: qualitative multiplex and quantitative real-time.
abstract: OBJECTIVE
  To establish a two-step polymerase chain reaction (PCR) diagnostic system for ocular toxoplasmosis.
  
  
  METHODS
  A total of 13 ocular fluid samples (11 aqueous humor and 2 vitreous fluid) were collected from 13 patients with clinically suspected ocular toxoplasmosis. Ten ocular samples from other uveitis patients and 20 samples from subjects without ocular inflammation were used as controls. Two polymerase chain reaction (PCR) methods, i.e., qualitative multiplex PCR and quantitative real-time PCR, were used to measure the toxoplasma genome (T. gondii B1 gene).
  
  
  RESULTS
  Qualitative multiplex PCR detected T. gondii B1 gene in the ocular fluids of 11 out of 13 patients with clinically suspected ocular toxoplasmosis. In real-time PCR, we detected high copy numbers of T. gondii DNA (5.1 × 10(2)-2.1 × 10(6) copies/mL) in a total of 10 patients (10/13, 77%). Only ocular toxoplasmosis scar lesions were observed in the three real-time PCR-negative patients. PCR assay results for the samples from the two control groups were all negative.
  
  
  CONCLUSIONS
  The two-step PCR examination to detect toxoplasma DNA is a useful tool for diagnosing ocular toxoplasmosis.
authors.affiliation: Department of Ophthalmology and Visual Science, Tokyo Medical and Dental University Graduate School of Medicine, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan. sunaoph@tmd.ac.jp.
authors.forename: Sunao
authors.initials: S
authors.lastname: Sugita
authors.affiliation: Department of Ophthalmology and Visual Science, Tokyo Medical and Dental University Graduate School of Medicine, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan.
authors.forename: Manabu
authors.initials: M
authors.lastname: Ogawa
authors.affiliation: Department of Ophthalmology and Visual Science, Tokyo Medical and Dental University Graduate School of Medicine, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan.
authors.forename: Shizu
authors.initials: S
authors.lastname: Inoue
authors.affiliation: Department of Virology, Medical Research Institute, Tokyo Medical and Dental University Graduate School of Medicine and Dental Sciences, Tokyo, Japan.
authors.forename: Norio
authors.initials: N
authors.lastname: Shimizu
authors.affiliation: Department of Ophthalmology and Visual Science, Tokyo Medical and Dental University Graduate School of Medicine, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan.
authors.forename: Manabu
authors.initials: M
authors.lastname: Mochizuki
mesh_terms: D000328:Adult; D000368:Aged; D001082:Aqueous Humor / Q000469:parasitology*; D017931:DNA Primers / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry; D016054:DNA, Protozoan / Q000032:analysis; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D018503:Genome, Protozoan / Q000235:genetics; D006564:Herpesviridae / Q000235:genetics; D006801:Humans; D008297:Male; D008875:Middle Aged; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D014122:Toxoplasma / Q000235:genetics / Q000302:isolation & purification*; D014126:Toxoplasmosis, Ocular / Q000175:diagnosis*; D014822:Vitreous Body / Q000469:parasitology*
chemical_list: D017931:DNA Primers; D015342:DNA Probes; D016054:DNA, Protozoan; D004279:DNA, Viral