title: Establishment of Multiplex Solid-Phase Strip PCR Test for Detection of 24 Ocular Infectious Disease Pathogens.
abstract: To establish and evaluate a new multiplex solid-phase strip polymerase chain reaction (strip PCR) for concurrent detection of common ocular infectious disease pathogens.
  
  
  
  A new multiplex strip PCR was established to detect 24 common ocular infectious disease pathogens: herpes simplex virus (HSV) 1, HSV2, varicella-zoster virus (VZV), Epstein-Barr virus (EBV), cytomegalovirus (CMV), human herpes virus (HHV) 6, HHV7, HHV8, human T-cell lymphotropic virus (HTLV)-1, adenovirus, Mycobacterium tuberculosis, Treponema pallidum, Propionibacterium acnes (P. acnes), bacterial 16S ribosomal RNA (rRNA), Candida species (Candida sp.), C. glabrata, C. krusei, Aspergillus, Fusarium, fungal 28S rRNA, Toxoplasma (T. gondii), Toxocara, Chlamydia trachomatis (C. trachomatis), and Acanthamoeba. Strip PCR was tested with a negative control (distilled water) and standard positive control DNA. Cutoffs of quantification cycle (Cq) values were determined with noninfectious ocular samples to avoid false-positives caused by contamination with P. acnes, bacterial 16S, and fungal 28S from reagents and ocular surfaces. A pilot study to evaluate the strip PCR was performed using infectious ocular samples (aqueous humor, vitreous, cornea, and tears) by strip PCR and previously developed capillary-type multiplex PCR and quantitative real-time PCR (qPCR).
  
  
  
  Strip PCR was verified with negative and positive controls. Strip PCR rapidly detected HSV1, HSV2, VZV, EBV, CMV, HHV6, HHV7, HTLV-1, adenovirus, P. acnes, bacterial 16S, Candida sp., C. glabrata, Aspergillus, fungal 28S, T. gondii, C. trachomatis, and Acanthamoeba in patient samples. The sensitivity was comparable to that of qPCR.
  
  
  
  Our novel strip PCR assay is a simple, rapid, and high-sensitivity method for detecting ocular infectious disease pathogens.
authors.affiliation: Department of Ophthalmology, Oita University, Oita, Japan.
authors.forename: Satoko
authors.initials: S
authors.lastname: Nakano
authors.affiliation: Laboratory for Retinal Regeneration, RIKEN Center for Developmental Biology, Kobe, Japan.
authors.forename: Sunao
authors.initials: S
authors.lastname: Sugita
authors.affiliation: Center for Stem Cell and Regenerative Medicine, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Tomaru
authors.affiliation: Laboratory for Retinal Regeneration, RIKEN Center for Developmental Biology, Kobe, Japan.
authors.forename: Ayumi
authors.initials: A
authors.lastname: Hono
authors.affiliation: Department of Ophthalmology, Oita University, Oita, Japan.
authors.forename: Takako
authors.initials: T
authors.lastname: Nakamuro
authors.affiliation: Department of Ophthalmology, Oita University, Oita, Japan.
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Kubota
authors.affiliation: Department of Ophthalmology & Visual Science, Tokyo Medical and Dental University, Graduate School of Medicine and Dental Sciences, Tokyo, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Takase
authors.affiliation: Department of Ophthalmology & Visual Science, Tokyo Medical and Dental University, Graduate School of Medicine and Dental Sciences, Tokyo, Japan 5Miyata Eye Hospital, Miyakonojo, Japan.
authors.forename: Manabu
authors.initials: M
authors.lastname: Mochizuki
authors.affiliation: Laboratory for Retinal Regeneration, RIKEN Center for Developmental Biology, Kobe, Japan.
authors.forename: Masayo
authors.initials: M
authors.lastname: Takahashi
authors.affiliation: Center for Stem Cell and Regenerative Medicine, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Norio
authors.initials: N
authors.lastname: Shimizu
mesh_terms: D000818:Animals; D001082:Aqueous Humor / Q000737:chemistry*; D001419:Bacteria / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis*; D004271:DNA, Fungal / Q000032:analysis*; D016054:DNA, Protozoan / Q000032:analysis*; D004279:DNA, Viral / Q000032:analysis*; D015817:Eye Infections / Q000175:diagnosis* / Q000382:microbiology; D005658:Fungi / Q000235:genetics; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D010271:Parasites / Q000235:genetics; D010865:Pilot Projects; D015203:Reproducibility of Results; D014780:Viruses / Q000235:genetics; D014822:Vitreous Body / Q000737:chemistry
chemical_list: D004269:DNA, Bacterial; D004271:DNA, Fungal; D016054:DNA, Protozoan; D004279:DNA, Viral