title: Simple Identification of Human Taenia Species by Multiplex Loop-Mediated Isothermal Amplification in Combination with Dot Enzyme-Linked Immunosorbent Assay.
abstract: For differential detection of Taenia solium, Taenia saginata, and Taenia asiatica, loop-mediated isothermal amplification (LAMP) assay targeting the cytochrome c oxidase subunit 1 gene has been recently developed and shown to be sensitive, specific, and effective. However, to achieve differential identification, one specimen requires three reaction mixtures containing a primer set of each Taenia species separately, which is complex and time consuming and increases the risk of cross-contamination. In this study, we developed a simple differential identification of human Taenia species using multiplex LAMP (mLAMP) in combination with dot enzyme-linked immunosorbent assay (dot-ELISA). Forward inner primers of T. solium, T. saginata, and T. asiatica labeled with fluorescein isothiocyanate (FITC), digoxigenin (DIG), and tetramethylrhodamine (TAMRA), respectively, and biotin-labeled backward inner primers were used in mLAMP. The mLAMP assay succeeded in specific amplification of each respective target gene in a single tube. Furthermore, the mLAMP product from each species was easily distinguished by dot-ELISA with an antibody specific for FITC, DIG, or TAMRA. The mLAMP assay in combination with dot-ELISA will make identification of human Taenia species simpler, easier, and more practical.
authors.affiliation: Department of Parasitology, Asahikawa Medical University, Asahikawa, Japan; Medical Research Centre, Institute of Medical Research and Medicinal Plants Studies (IMPM), Ministry of Scientific Research and Innovation, Yaoundé, Cameroon; Center for Human Evolution Modeling Research, Primate Research Institute, Kyoto University, Inuyama, Japan ankouawa@gmail.com.
authors.forename: Agathe
authors.initials: A
authors.lastname: Nkouawa
authors.affiliation: Department of Parasitology, Asahikawa Medical University, Asahikawa, Japan; Medical Research Centre, Institute of Medical Research and Medicinal Plants Studies (IMPM), Ministry of Scientific Research and Innovation, Yaoundé, Cameroon; Center for Human Evolution Modeling Research, Primate Research Institute, Kyoto University, Inuyama, Japan.
authors.forename: Yasuhito
authors.initials: Y
authors.lastname: Sako
authors.affiliation: Department of Parasitology, Asahikawa Medical University, Asahikawa, Japan; Medical Research Centre, Institute of Medical Research and Medicinal Plants Studies (IMPM), Ministry of Scientific Research and Innovation, Yaoundé, Cameroon; Center for Human Evolution Modeling Research, Primate Research Institute, Kyoto University, Inuyama, Japan.
authors.forename: Munehiro
authors.initials: M
authors.lastname: Okamoto
authors.affiliation: Department of Parasitology, Asahikawa Medical University, Asahikawa, Japan; Medical Research Centre, Institute of Medical Research and Medicinal Plants Studies (IMPM), Ministry of Scientific Research and Innovation, Yaoundé, Cameroon; Center for Human Evolution Modeling Research, Primate Research Institute, Kyoto University, Inuyama, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Ito
mesh_terms: D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D003576:Electron Transport Complex IV / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques; D012680:Sensitivity and Specificity; D013621:Taenia / Q000145:classification* / Q000235:genetics; D013622:Taeniasis / Q000469:parasitology*
chemical_list: D004247:DNA; D003576:Electron Transport Complex IV