title: Use of a comprehensive polymerase chain reaction system for diagnosis of ocular infectious diseases.
abstract: OBJECTIVE
  To measure the genomic DNA of ocular infectious pathogens in ocular fluids and to analyze the clinical relevance of these pathogens in uveitis and endophthalmitis.
  
  
  METHODS
  Prospective clinical case series.
  
  
  METHODS
  A total of 500 patients with infectious uveitis and endophthalmitis were examined at Tokyo Medical and Dental University, Tokyo Medical University, Kyushu University, Osaka University, and Kyoto Prefectural University, all in Japan.
  
  
  METHODS
  Genomic DNA of bacteria, fungi, parasites, and viruses in collected intraocular samples were examined by comprehensive polymerase chain reaction (PCR). Samples were analyzed first by multiplex PCR and quantitative real-time PCR for human herpes viruses (HHVs) 1 through 8 and toxoplasma. Subsequently, samples were examined by broad-range real-time PCR for bacterial 16S and fungal 18S/28S ribosomal DNA (rDNA).
  
  
  METHODS
  Infectious uveitis and endophthalmitis diagnoses were obtained when using the PCR system. Calculations of the positivity and the diagnostic parameters such as sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) also were evaluated.
  
  
  RESULTS
  In all of the tested infectious uveitis and endophthalmitis patients, either herpes simplex virus type 1 (n = 18), herpes simplex virus type 2 (n = 4), varicella-zoster virus (n = 55), Epstein-Barr virus (n = 17), cytomegalovirus (n = 68), HHV type 6 (n = 2), toxoplasma (n = 6), bacterial 16S (n = 33), or fungal 18S/28S (n = 11) genome was detected. Neither HHV type 7 nor HHV type 8 DNA was detected in any of the samples. Of the 21 false-negative results found during the PCR analyses, 12 cases were negative for patients clinically suspected of having bacterial endophthalmitis. Conversely, false-positive results for the comprehensive PCR examinations occurred in only 3 cases that subsequently were found to have bacterial 16S rDNA. Diagnostic parameters for the sensitivity, specificity, PPV, and NPV of our PCR examinations were 91.3%, 98.8%, 98.6%, and 92.4%, respectively.
  
  
  CONCLUSIONS
  Use of our comprehensive PCR assay to examine ocular samples in patients with endophthalmitis and uveitis seems to be clinically useful for detecting infectious antigen DNA. Thus, this PCR method is a reliable tool for both diagnosing ocular disorders and further screening of patients for intraocular infections.
  
  
  BACKGROUND
  The author(s) have no proprietary or commercial interest in any materials discussed in this article.
authors.affiliation: Department of Ophthalmology & Visual Science, Tokyo Medical and Dental University, Tokyo, Japan. sunaoph@cdb.riken.jp
authors.forename: Sunao
authors.initials: S
authors.lastname: Sugita
authors.forename: Manabu
authors.initials: M
authors.lastname: Ogawa
authors.forename: Norio
authors.initials: N
authors.lastname: Shimizu
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Morio
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Ohguro
authors.forename: Kei
authors.initials: K
authors.lastname: Nakai
authors.forename: Kazuichi
authors.initials: K
authors.lastname: Maruyama
authors.forename: Kenji
authors.initials: K
authors.lastname: Nagata
authors.forename: Atsunobu
authors.initials: A
authors.lastname: Takeda
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Usui
authors.forename: Koh-Hei
authors.initials: KH
authors.lastname: Sonoda
authors.forename: Masaru
authors.initials: M
authors.lastname: Takeuchi
authors.forename: Manabu
authors.initials: M
authors.lastname: Mochizuki
mesh_terms: D004269:DNA, Bacterial / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D009877:Endophthalmitis / Q000175:diagnosis* / Q000382:microbiology / Q000469:parasitology / Q000821:virology; D015817:Eye Infections / Q000175:diagnosis* / Q000382:microbiology / Q000469:parasitology / Q000821:virology; D005189:False Positive Reactions; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D011446:Prospective Studies; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012339:RNA, Ribosomal, 28S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D018139:Simplexvirus / Q000235:genetics; D014122:Toxoplasma / Q000235:genetics; D014605:Uveitis / Q000175:diagnosis* / Q000382:microbiology / Q000469:parasitology / Q000821:virology
chemical_list: D004269:DNA, Bacterial; D004271:DNA, Fungal; D016054:DNA, Protozoan; D004275:DNA, Ribosomal; D004279:DNA, Viral; D012337:RNA, Ribosomal, 18S; D012339:RNA, Ribosomal, 28S