title: Application of environmental DNA analysis for the detection of Opisthorchis viverrini DNA in water samples.
abstract: Opisthorchiasis, which can lead to cholangiocarcinoma in cases of chronic infection, is a major public health problem in Southeast Asian countries. The trematode, Opisthorchis viverrini, is the causative agent of the disease. Accurate and rapid monitoring of O. viverrini is crucial for disease prevention and containment. Therefore, in this study we sought to develop a novel species-specific real-time PCR assay for detecting 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 in Lao PDR (21 and 19 samples were positive by real-time PCR and traditional PCR, respectively). The efficacy of eDNA analysis and its applicability in the field were tested using a total of 94 environmental water samples collected from 44 sites in Savannakhet, Lao PDR during May and October 2015 and February 2016. O. viverrini eDNA was detected in five samples by real-time PCR, indicating the presence of the fluke in the area and the risk of infection for individuals consuming fish from these water sources. The application of eDNA analysis would facilitate the identification of O. viverrini endemic hotspots and contribute to the ecological control of opisthorchiasis, and this strategy can be applied to other eukaryotic water pathogens.
authors.affiliation: Graduate School of Human Development and Environment, Kobe University, 3-11, Tsurukabuto, Nada-ku, Kobe 657-8501 Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Hashizume
authors.affiliation: Graduate School of Health Sciences, Niigata University, 2-746 Asahimachi-dori, Chuo-ku, Niigata 951-8518, Japan.
authors.forename: Megumi
authors.initials: M
authors.lastname: Sato
authors.affiliation: Department of Tropical Medicine and Parasitology, Dokkyo Medical University, 880 Kitakobayashi, Mibu-machi, Shimotsuga-gun, Tochigi 321-0293, Japan.
authors.forename: Marcello Otake
authors.initials: MO
authors.lastname: Sato
authors.affiliation: Graduate School of Health Sciences, Niigata University, 2-746 Asahimachi-dori, Chuo-ku, Niigata 951-8518, Japan.
authors.forename: Sumire
authors.initials: S
authors.lastname: Ikeda
authors.affiliation: Department of Helminthology, Faculty of Tropical Medicine, Mahidol University, 420/6 Ratchawithi Road, Ratchathewi, Bangkok 10400, Thailand.
authors.forename: Tippayarat
authors.initials: T
authors.lastname: Yoonuan
authors.affiliation: Department of Helminthology, Faculty of Tropical Medicine, Mahidol University, 420/6 Ratchawithi Road, Ratchathewi, Bangkok 10400, Thailand.
authors.forename: Surapol
authors.initials: S
authors.lastname: Sanguankiat
authors.affiliation: Savannakhet Provincial Health Department, Thahea village, Kaysone District, Savannakhet Province, Lao Democratic People's Republic.
authors.forename: Tiengkham
authors.initials: T
authors.lastname: Pongvongsa
authors.affiliation: School of Tropical Medicine and Global Health, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan.
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Moji
authors.affiliation: Graduate School of Human Development and Environment, Kobe University, 3-11, Tsurukabuto, Nada-ku, Kobe 657-8501 Japan. Electronic address: minamoto@people.kobe-u.ac.jp.
authors.forename: Toshifumi
authors.initials: T
authors.lastname: Minamoto
mesh_terms: D000818:Animals; D018281:Cholangiocarcinoma / Q000469:parasitology; D018509:DNA, Helminth / Q000032:analysis / Q000235:genetics; D004784:Environmental Monitoring / Q000379:methods*; D005399:Fishes / Q000469:parasitology; D009889:Opisthorchiasis / Q000469:parasitology; D009891:Opisthorchis / Q000235:genetics* / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction; D017747:Seafood / Q000469:parasitology; D013045:Species Specificity; D014867:Water / Q000469:parasitology*
chemical_list: D018509:DNA, Helminth; D014867:Water
keywords: Environmental DNA (eDNA); Lao PDR; Liver fluke; Opisthorchis viverrini; Real-time PCR