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Novel diagnosis of fungal endophthalmitis by broad-range real-time PCR detection of fungal 28S ribosomal DNA

  • Inflammatory Disorders
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Abstract

Aim

To detect the fungal genome in the ocular fluids of patients with fungal endophthalmitis by using a novel broad-range polymerase chain reaction (PCR) system.

Methods

After informed consent was obtained, ocular fluid samples (aqueous humor or vitreous fluids) were collected from 497 patients (76 patients with infectious endophthalmitis including clinically suspected bacterial and fungal endophthalmitis and 421 patients with infectious or non-infectious uveitis). Forty ocular samples from non-infectious patients without ocular inflammation were collected as controls. Fungal ribosomal DNA (28 S rDNA) was measured by a quantitative real-time PCR assay.

Results

Fungal 28 S rDNA of the major fungal species, such as Candida, Aspergillus, and Cryptococcus, were detected by novel broad-range real-time PCR examination (>101 copies/ml). Fungal 28 S rDNA was detected in the ocular fluids of 11 patients with endophthalmitis or uveitis (11/497, 2.2%). All 11 positive samples were detected in the infectious endophthalmitis patients (11/76, 14.5%). These PCR-positive ocular fluids had high copy numbers of fungal 28 S rDNA (range, 1.7 × 103 to 7.9 × 106 copies/ml), which indicated the presence of fungal infection. Of the 11 patients who were PCR positive, further examinations led to a diagnosis of fungal endophthalmitis in ten patients. The fungal 28 S rDNA was detected in one non-infectious case (a false-positive case). In addition, there were two PCR false-negative cases that were clinically suspected of having fungal endophthalmitis.

Conclusions

This novel quantitative broad-range PCR of fungal 28 S rDNA is a useful tool for diagnosing endophthalmitis related to fungal infections.

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Acknowledgments

We greatly appreciate the expert technical assistance of Ikuyo Yamamoto and Chizuru Kato.

Funding

This work was supported by Comprehensive Research on Disability, Health and Welfare, Health and Labour Sciences Research Grants, Ministry Health, Labour and Welfare, Japan.

Competing interests

None.

Contributors

MO was the principal investigator, designed and performed experiments, and wrote the manuscript. SS designed and conceptualized the study and drafted and edited the manuscript. KW and NS performed PCR assays. MM designed and conceptualized the study and edited the manuscript.

Data sharing statement

No additional data.

Ethics approval

Ethics approval was provided by the Institutional Ethics Committee of Tokyo Medical and Dental University.

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Authors and Affiliations

  1. Department of Ophthalmology & Visual Science, Tokyo Medical and Dental University Graduate School of Medicine, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan

    Manabu Ogawa, Sunao Sugita & Manabu Mochizuki

  2. Department of Virology, Division of Medical Science, Tokyo Medical and Dental University, Graduate School of Medical and Dental Sciences, Tokyo, Japan

    Ken Watanabe & Norio Shimizu

Authors
  1. Manabu Ogawa
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  2. Sunao Sugita
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  3. Ken Watanabe
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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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Ogawa, M., Sugita, S., Watanabe, K. et al. Novel diagnosis of fungal endophthalmitis by broad-range real-time PCR detection of fungal 28S ribosomal DNA. Graefes Arch Clin Exp Ophthalmol 250, 1877–1883 (2012). https://doi.org/10.1007/s00417-012-2015-7

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  • DOI: https://doi.org/10.1007/s00417-012-2015-7

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