July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Evaluation of a multiplex Strip PCR examination for infectious uveitis and endophthalmitis: A prospective multi-center study
Author Affiliations & Notes
  • Satoko Nakano
    Ophthalmology, Oita University, Yufu-City, Oita, Japan
  • Yasuhiro Tomaru
    Center for Stem Cell and Regenerative Medicine, Tokyo Medical and Dental University, Tokyo, Japan
  • Hiroshi Takase
    Department of Ophthalmology & Visual Science, Tokyo Medical and Dental University Graduate School of Medicine and Dental Sciences, Tokyo, Japan
  • Toshiaki Kubota
    Ophthalmology, Oita University, Yufu-City, Oita, Japan
  • Manabu Mochizuki
    Department of Ophthalmology & Visual Science, Tokyo Medical and Dental University Graduate School of Medicine and Dental Sciences, Tokyo, Japan
  • Norio Shimizu
    Center for Stem Cell and Regenerative Medicine, Tokyo Medical and Dental University, Tokyo, Japan
  • Sunao Sugita
    Laboratory for Retinal Regeneration, RIKEN Center for Developmental Biology, Kobe, Japan
  • Footnotes
    Commercial Relationships   Satoko Nakano, None; Yasuhiro Tomaru, None; Hiroshi Takase, None; Toshiaki Kubota, None; Manabu Mochizuki, None; Norio Shimizu, None; Sunao Sugita, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 4170. doi:
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      Satoko Nakano, Yasuhiro Tomaru, Hiroshi Takase, Toshiaki Kubota, Manabu Mochizuki, Norio Shimizu, Sunao Sugita; Evaluation of a multiplex Strip PCR examination for infectious uveitis and endophthalmitis: A prospective multi-center study. Invest. Ophthalmol. Vis. Sci. 2018;59(9):4170.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : To evaluate our multiplex Strip PCR test based on 24 common pathogens of ocular infectious diseases. To find optimal pathogens’ combination of Strip PCR for detecting infectious uveitis and endophthalmitis.

Methods : Multi-center prospective evaluation of the diagnostic PCR test was conducted. The 616 subjects (patients with infectious uveitis, endophthalmitis, and controls) were examined at 14 institutes in Japan. The Strip PCR test includes herpes simplex virus (HSV) 1–8, human T-cell lymphotropic virus (HTLV)-1, adenovirus, Toxoplasma gondii (T. gondii), Chlamydia trachomatis, Acanthamoeba, Toxocara, Mycobacterium tuberculosis, Propionibacterium acnes (P. acnes), Treponema pallidum (T. pallidum), bacterial 16S ribosomal (r)RNA, Candida sp., Candida glabrata, Candida krusei, Fusarium, Aspergillus, and fungal 28S rRNA. Genomic DNA was collected from the aqueous humor and vitreous fluid and analyzed by Strip PCR and quantitative real-time PCR (qPCR). Results of Strip PCR were compared to those of clinical diagnosis and qPCR. Main Outcome Measures, sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated.

Results : We observed high sensitivity (93.5%), specificity (97.9%), PPV (98.1%), and NPV (92.9%) in Strip PCR, and it detected DNA for primary pathogenesis and mixed infections. Its quantification cycle value (Cq) varied according to diseases’ severity. Some of viruses such as HSV1, HSV2, varicella-zoster virus (VZV), Epstein-Barr virus (EBV), human herpes virus 6 (HHV6), and HTLV-1 showed 100% PPV and NPV. Strip PCR and qPCR failed to detect Toxocara and M. tuberculosis. Only five false-positives were obtained for P. acnes and bacterial 16S, which normally live in ocular surface and reagents in PCR. The diagnostic parameters of the Strip PCR reconfigured specifically for infectious uveitis, with HSV1, HSV2, VZV, HTLV-1, HHV6, EBV, CMV, T. gondii, and T. pallidum were 100.0% PPV, 99.2% NPV, 99.2% sensitivity, and 100.0% specificity. Strip PCR also had good correlation between Cq values and DNA copy numbers from qPCR.

Conclusions : Strip PCR exhibited high diagnostic parameters and this allows accurate etiological and exclusive diagnosis of infectious uveitis and endophthalmitis for adequate treatments.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

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