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Real-time PCR method for the detection and quantification of Acanthamoeba species in various types of water samples

Parasitology Research volume 112, pages 1131–1136 (2013)Cite this article

Abstract

In this study, a quantitative real-time PCR was developed to detect and quantify Acanthamoeba spp. in various environmental water samples. The water samples were taken from watershed, water treatment plant, and three thermal spring recreation areas. The overall detection rate was 14.2 % (25/176) for Acanthamoeba spp. The percentages of samples containing Acanthamoeba spp. from river water, raw drinking water, and thermal spring water were 13 % (13/100), 25 % (7/28), and 10.4 % (5/48), respectively. Acanthamoeba spp. concentrations were determined according to SYBR Green quantitative real-time PCR. A plasmid-based standard curve was constructed to determine the Acanthamoeba concentration using dilution factors for achieving 1.36 × 109 gene copies per PCR for 18S rRNA gene in Acanthamoeba spp. The resulting concentrations varied by the type of water, in the range of 46–2.6 × 102 cells/l in positive raw drinking water, 2.7 × 102–1.5 × 104 cells/l in river water, and 54–1.7 × 103 cells/l in thermal spring water. The presence of Acanthamoeba spp. in the raw drinking water samples was also found to have a significant difference with heterotrophic plate count. The presence of Acanthamoeba spp. in various aquatic environments may be a potential health hazard and must be further evaluated.

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Acknowledgments

This work was supported by a research grant from the National Science Council of Taiwan, Republic of China (NSC 100-2116-M-194-004-MY2).

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Affiliations

  1. Department of Earth and Environmental Sciences, National Chung Cheng University, Chiayi, Taiwan, Republic of China

    Po-Min Kao, Bing-Mu Hsu, Cheng-Yu She & Shu-Min Shen

  2. Department of Surgery, Tungs’ Taichung MetroHarbor Hospital, Taichung, Taiwan, Republic of China

    Min-Che Tung

  3. Department of Emergency Medicine, Cheng Hsin Hospital, Taipei, Taiwan, Republic of China

    Hsien-Lung Tsai

  4. Department of Surgery, Mackay Memorial Hospital, Mackay Medicine, Nursing, and Management College, Taipei, Taiwan, Republic of China

    Wen-Chien Huang

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  1. Po-Min Kao
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  2. Min-Che Tung
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  3. Bing-Mu Hsu
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  4. Hsien-Lung Tsai
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  5. Cheng-Yu She
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  6. Shu-Min Shen
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  7. Wen-Chien Huang
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Correspondence to Min-Che Tung or Bing-Mu Hsu.

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Hsien-Lung Tsai equally contributed with the first author.

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Kao, PM., Tung, MC., Hsu, BM. et al. Real-time PCR method for the detection and quantification of Acanthamoeba species in various types of water samples. Parasitol Res 112, 1131–1136 (2013). https://doi.org/10.1007/s00436-012-3242-x

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  • DOI: https://doi.org/10.1007/s00436-012-3242-x

Keywords

  • Water Quality Parameter
  • River Water Sample
  • Heterotrophic Plate Count
  • Acanthamoeba Keratitis
  • Thermal Spring Water