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Diagnosis of ocular toxoplasmosis by two polymerase chain reaction (PCR) examinations: qualitative multiplex and quantitative real-time

  • Clinical Investigation
  • Published:
Japanese Journal of Ophthalmology Aims and scope Submit manuscript

Abstract

Aim

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 × 102–2.1 × 106 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.

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Acknowledgments

Drs Kazuichi Maruyama and Kenji Nagata of the Department of Ophthalmology, Kyoto Prefectural University of Medicine, collected and sent the samples used in this study. We are grateful for the expert technical assistance of Mr Ken Watanabe. This work was supported by Comprehensive Research on Disability, Health and Welfare, Health and Labour Sciences Research Grants, Ministry Health, Labour and Welfare, Japan.

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Authors and Affiliations

  1. 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

    Sunao Sugita, Manabu Ogawa, Shizu Inoue & Manabu Mochizuki

  2. Department of Virology, Medical Research Institute, Tokyo Medical and Dental University Graduate School of Medicine and Dental Sciences, Tokyo, Japan

    Norio Shimizu

Authors
  1. Sunao Sugita
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  2. Manabu Ogawa
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  3. Shizu Inoue
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  4. Norio Shimizu
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  5. Manabu Mochizuki
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Corresponding author

Correspondence to Sunao Sugita.

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Sugita, S., Ogawa, M., Inoue, S. et al. Diagnosis of ocular toxoplasmosis by two polymerase chain reaction (PCR) examinations: qualitative multiplex and quantitative real-time. Jpn J Ophthalmol 55, 495–501 (2011). https://doi.org/10.1007/s10384-011-0065-8

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  • DOI: https://doi.org/10.1007/s10384-011-0065-8

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