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KMID : 0364820220580010033
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Korean Journal of Microbiology
2022 Volume.58 No. 1 p.33 ~ p.40
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Optimization and application of multiplex nested PCR to detect free-living amoebas in biomass-rich freshwater samples
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Kim Eun-Jeong
Shin Ji-Soo
Kim Do-Hwan
Cho Young-Cheol
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Abstract
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Multiplex nested PCR method to detect Naegleria fowleri and Acanthamoeba spp., opportunistic pathogenic free-living amoebas (FLAs), in biomass-rich samples was optimized and applied in the samples taken from Korean freshwaters. The appropriate PCR primers targeted 18S rRNA gene of these FLAs were selected or designed and the reaction conditions were optimized. The optimized method can detect these FLAs even if a single cell is present in the sample. When the optimized method was applied to the 56 samples taken from the Nakdong River in Korea, N. fowleri and Acanthamoeba spp. were detected in 9 samples (16.1%) and 55 samples (98.2%), respectively. In conclusion the method optimized in this study can be used to monitor N. fowleri and Acanthamoeba spp. and in biomass-rich samples.
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KEYWORD
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Naegleria fowleri, Acanthamoeba spp., biomass-rich samples, multiplex nested PCR, pathogenic free-living amoeba
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