Parasitology Research

, Volume 112, Issue 3, pp 1131–1136 | Cite as

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

  • Po-Min Kao
  • Min-Che TungEmail author
  • Bing-Mu HsuEmail author
  • Hsien-Lung Tsai
  • Cheng-Yu She
  • Shu-Min Shen
  • Wen-Chien Huang
Original Paper


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.


Water Quality Parameter River Water Sample Heterotrophic Plate Count Acanthamoeba Keratitis Thermal Spring Water 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



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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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Po-Min Kao
    • 1
  • Min-Che Tung
    • 2
    Email author
  • Bing-Mu Hsu
    • 1
    Email author
  • Hsien-Lung Tsai
    • 3
  • Cheng-Yu She
    • 1
  • Shu-Min Shen
    • 1
  • Wen-Chien Huang
    • 4
  1. 1.Department of Earth and Environmental SciencesNational Chung Cheng UniversityChiayiRepublic of China
  2. 2.Department of SurgeryTungs’ Taichung MetroHarbor HospitalTaichungRepublic of China
  3. 3.Department of Emergency MedicineCheng Hsin HospitalTaipeiRepublic of China
  4. 4.Department of Surgery, Mackay Memorial HospitalMackay Medicine, Nursing, and Management CollegeTaipeiRepublic of China

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