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Detection of Candida and Aspergillus species DNA using broad-range real-time PCR for fungal endophthalmitis

  • Inflammatory Disorders
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Abstract

Background

The goal of this work is to establish a broad-range real-time polymerase chain reaction (PCR) diagnostic system for ocular fungal infection and to measure Candida and Aspergillus DNA in the ocular fluids obtained from unknown uveitis/endophthalmitis patients.

Methods

After obtaining informed consent, intraocular fluids (aqueous humor and vitreous fluid samples) were collected from 54 patients with idiopathic uveitis or endophthalmitis. Samples were assayed for Candida or Aspergillus DNA using broad-range (18S rRNA sequences) quantitative real-time PCR.

Results

Candida or Aspergillus DNA was detected in seven out of 54 patient ocular samples (13%). These PCR-positive samples showed significantly high copy numbers of Candida or Aspergillus DNA. On the other hand, fungal DNA was not detected in any of the other 46 samples collected from these idiopathic uveitis or endophthalmitis patients. In the one PCR-negative case, PCR did not detect any fungal genome in the sample, even though this patient was clinically suspected of having Candida endophthalmitis. Real-time PCR results were negative for fungal DNA in the bacterial endophthalmitis patients and in various uveitis patients. In addition, fungal DNA was also not detected in patients without ocular inflammation (controls).

Conclusions

Analysis of ocular samples by this broad-range real-time PCR method can be utilized for rapid diagnosis of patients suffering from unknown intraocular disorders such as idiopathic uveitis/endophthalmitis.

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Acknowledgements

We thank Ms. Miki Katayama and Shizu Inoue for their technical assistance. We would like to also thank Drs. Hiroshi Takase and Yoshiharu Sugamoto for obtaining the samples used in this study. This work was supported by a Comprehensive Research on Disability, Health and Welfare grant, along with a Health and Labour Sciences Research Grant from the Ministry of Health, Labour and Welfare, Japan.

No financial relationships exist in the publishing of this work.

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Authors and Affiliations

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

    Sunao Sugita, Koju Kamoi, Manabu Ogawa & 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. Sunao Sugita
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  2. Koju Kamoi
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  3. Manabu Ogawa
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  4. Ken Watanabe
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  5. Norio Shimizu
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  6. Manabu Mochizuki
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Corresponding author

Correspondence to Sunao Sugita.

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Sugita, S., Kamoi, K., Ogawa, M. et al. Detection of Candida and Aspergillus species DNA using broad-range real-time PCR for fungal endophthalmitis. Graefes Arch Clin Exp Ophthalmol 250, 391–398 (2012). https://doi.org/10.1007/s00417-011-1819-1

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  • DOI: https://doi.org/10.1007/s00417-011-1819-1

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