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KMID : 0364820180540020098
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Korean Journal of Microbiology
2018 Volume.54 No. 2 p.98 ~ p.104
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Surveillance of Acanthamoeba spp. and Naegleria fowleri in environmental water by using the duplex real-time PCR
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Kim Min-Jeong
Lee Gyu-Cheol
Kim Kun-Woo
Lee Hyun-Ji
Kim Min-Young
Seo Dae-Keun
Lee Jeong-Yeob
Cho Young-Cheol
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Abstract
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Naegleria fowleri and Acanthamoeba spp. are free-living amoebasthat are widely distributed in natural environments. Althoughuncommon, infection with these protozoans can cause fataldisease in humans and animals. In this study, in order to select theappropriate method to survey Naegleria fowleri and Acanthamoebaspp. in water samples, four molecular biology techniques andone commercially available kit for real-time PCR were compared. The results indicated that the duplex real-time PCR was themost sensitive, and could be used to simultaneously detect twodifferent free-living amoebas. Using the duplex real-time PCRapproach, the two free-living amoebas were surveyed in threelocal streams in Daejeon, Republic of Korea. The concentratedfree-living amoebas were inoculated onto non-nutrient agarplates which had been spread with heat-inactivated Escherichiacoli and incubated for 5~7 days. After incubation, gDNA wasextracted and used as the template for amplification by duplexreal-time PCR. Acanthamoeba spp. and N. fowleri was detectedfrom ten (83.3%) and two (16.6%) of the twelve samples,respectively. As these two free-living amoebas can be fatal,continuous surveillance is needed to track their distribution inthe aquatic environment for the drinking water safety.
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KEYWORD
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Acanthamoeba spp., Naegleria fowleri, duplex real-time PCR
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