Application of Realtime PCR and environmental DNA analysis for detecting minute flukes

Tran Thanh Duong; Nguyen Thu Huong; Nguyen Thi Hong Ngoc; Nguyen Thi Thanh Huyen; Nguyen Thi Huong Binh

Volume : VII, Issue : XI, November - 2018

Application of Realtime PCR and environmental DNA analysis for detecting minute flukes

Tran Thanh Duong, Nguyen Thu Huong, Nguyen Thi Hong Ngoc, Nguyen Thi Thanh Huyen, Nguyen Thi Huong Binh

Abstract :

Therefore, in this study we sought to develop a novel species-specific real-time PCR assay for detecting C. sennisis and or O. viverrini using environmental DNA (eDNA). The diagnostic sensitivity of the newly developed real-time PCR assay was similar to that of the traditional PCR assay for 50 fecal samples collected which were positive for small fluke egg were selected for adult and egg flukes in sedimentation feces. The efficacy of eDNA analysis and its applicability in the field were tested using a total of 120 environmental water samples collected from Bac Giang and Binh Dinh provinces in 2017. The results showed that C. sennisis and O. viverrini DNA was detected in 11 (22.0%) and 7 (14.0%) samples of sedimentation feces by real-time PCR. There were two of this pathogen eDNA in 2 (1.6%) and 4 (3.3%) water samples. The application of eDNA analysis would use the identification of fishborn-trematode endemic hotspots and contribute to the ecological control of closisenisis and opisthorchiasis, and this strategy can be applied to other eukaryotic environmental water pathogens