title: Rapid identification of nine species of diphyllobothriidean tapeworms by pyrosequencing.
abstract: The identification of diphyllobothriidean tapeworms (Cestoda: Diphyllobothriidea) that infect humans and intermediate/paratenic hosts is extremely difficult due to their morphological similarities, particularly in the case of Diphyllobothrium and Spirometra species. A pyrosequencing method for the molecular identification of pathogenic agents has recently been developed, but as of yet there have been no reports of pyrosequencing approaches that are able to discriminate among diphyllobothriidean species. This study, therefore, set out to establish a pyrosequencing method for differentiating among nine diphyllobothriidean species, Diphyllobothrium dendriticum, Diphyllobothrium ditremum, Diphyllobothrium latum, Diphyllobothrium nihonkaiense, Diphyllobothrium stemmacephalum, Diplogonoporus balaenopterae, Adenocephalus pacificus, Spirometra decipiens and Sparganum proliferum, based on the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene as a molecular marker. A region of 41 nucleotides in the cox1 gene served as a target, and variations in this region were used for identification using PCR plus pyrosequencing. This region contains nucleotide variations at 12 positions, which is enough for the identification of the selected nine species of diphyllobothriidean tapeworms. This method was found to be a reliable tool not only for species identification of diphyllobothriids, but also for epidemiological studies of cestodiasis caused by diphyllobothriidean tapeworms at public health units in endemic areas.
authors.affiliation: Faculty of Medicine, Mahasarakham University, Mahasarakham, 44000, Thailand.|Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen, 40002, Thailand.
authors.forename: Tongjit
authors.initials: T
authors.lastname: Thanchomnang
authors.affiliation: Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen, 40002, Thailand.|Division of Cell Biology, Department of Preclinical Sciences, Faculty of Medicine, Thammasat University, Rangsit Campus, Pathum Thani, 12120, Thailand.
authors.forename: Chairat
authors.initials: C
authors.lastname: Tantrawatpan
authors.affiliation: Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen, 40002, Thailand.|Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand.
authors.forename: Pewpan M
authors.initials: PM
authors.lastname: Intapan
authors.affiliation: Faculty of Medicine, Mahasarakham University, Mahasarakham, 44000, Thailand.|Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen, 40002, Thailand.|Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand.
authors.forename: Oranuch
authors.initials: O
authors.lastname: Sanpool
authors.affiliation: Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen, 40002, Thailand.|Department of Microbiology, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand.
authors.forename: Viraphong
authors.initials: V
authors.lastname: Lulitanond
authors.affiliation: Faculty of Medicine, Mahasarakham University, Mahasarakham, 44000, Thailand.
authors.forename: Somjintana
authors.initials: S
authors.lastname: Tourtip
authors.affiliation: Department of Parasitology, National Institute of Infectious Diseases, Tokyo, 162-8640, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yamasaki
authors.affiliation: Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen, 40002, Thailand.|Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand.
authors.forename: Wanchai
authors.initials: W
authors.lastname: Maleewong
mesh_terms: D000818:Animals; D003576:Electron Transport Complex IV / Q000235:genetics*; D015801:Helminth Proteins / Q000235:genetics*; D059014:High-Throughput Nucleotide Sequencing; D010986:Platyhelminths / Q000166:cytology / Q000235:genetics
chemical_list: D015801:Helminth Proteins; D003576:Electron Transport Complex IV