Real-time PCR method for the detection and quantification of Acanthamoeba species in various types of water samples
- 545 Downloads
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.
KeywordsWater Quality Parameter River Water Sample Heterotrophic Plate Count Acanthamoeba Keratitis Thermal Spring Water
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).
- Amorn L, Chantira S, Somchai B, Yaowalark S (2005) Free-living ameba contamination in natural hot springs in Thailand. Southeast Asian J Trop Med Public Health 36:1–5Google Scholar
- APHA (2005) Standard method for the examination of water and wastewater. APHA, WEF and AWWA, Washington, D.CGoogle Scholar
- Fields BS (1993) Legionella and protozoa: interaction of a pathogen and its natural host. In: Barbaree JM, Breiman RF, Dufour AP (eds) Legionella: current status and emerging perspectives. ASM, Washington, D.C., pp 70–72Google Scholar
- Lorenzo-Morales J, Ortega-Rivas A, Martínez E, Khoubbane M, Artigas P, Periago MV, Foronda P, Abreu-Acosta N, Valladares B, Mas-Coma S (2006) Acanthamoeba isolates belonging to T1, T2, T3, T4 and T7 genotypes from environmental freshwater samples in the Nile Delta region, Egypt. Acta Protozool 100:63–69Google Scholar