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However, discrimination errors have been reported in both methods. This study aimed to develop a multiplex PCR based on a novel nuclear marker, the fatty acid binding protein type I (<jats:italic>FABP<\/jats:italic>) type I gene.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Methods<\/jats:title>\n                <jats:p>Nucleotide sequence variations of FABP type I were analyzed using DNA samples of <jats:italic>F. hepatica<\/jats:italic>, <jats:italic>F. gigantica<\/jats:italic>, and hybrid <jats:italic>Fasciola<\/jats:italic> flukes obtained from 11 countries in Europe, Latin America, Africa, and Asia. A common forward primer for <jats:italic>F. hepatica<\/jats:italic> and <jats:italic>F. gigantica<\/jats:italic> and two specific reverse primers for <jats:italic>F. hepatica<\/jats:italic> and <jats:italic>F. gigantica<\/jats:italic> were designed for multiplex PCR.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>Specific fragments of <jats:italic>F. hepatica<\/jats:italic> (290\u00a0bp) and <jats:italic>F. gigantica<\/jats:italic> (190\u00a0bp) were successfully amplified using multiplex PCR. However, the hybrid flukes contained fragments of both species. The multiplex PCR for FABP\u00a0type I could precisely discriminate the 1312 <jats:italic>Fasciola<\/jats:italic> samples used in this study. Notably, no discrimination errors were observed with this novel method.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusions<\/jats:title>\n                <jats:p>Multiplex PCR for <jats:italic>FABP<\/jats:italic> type I can be used as a species discrimination marker in place of <jats:italic>pepck<\/jats:italic> and <jats:italic>pold<\/jats:italic>. The robustness of the species-specific primer should be continuously examined using a larger number of <jats:italic>Fasciola<\/jats:italic> flukes worldwide in the future since nucleotide substitutions in the primer regions may cause amplification errors.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Graphical abstract<\/jats:title>\n                \n              <\/jats:sec>","DOI":"10.1186\/s13071-022-05538-7","type":"journal-article","created":{"date-parts":[[2022,10,20]],"date-time":"2022-10-20T19:03:03Z","timestamp":1666292583000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Development of novel DNA marker for species discrimination of Fasciola flukes based on the fatty acid binding protein type I gene"],"prefix":"10.1186","volume":"15","author":[{"given":"Emi","family":"Okamoto","sequence":"first","affiliation":[]},{"given":"Michiyo","family":"Tashiro","sequence":"additional","affiliation":[]},{"given":"Pedro","family":"Ortiz","sequence":"additional","affiliation":[]},{"given":"Uday Kumar","family":"Mohanta","sequence":"additional","affiliation":[]},{"given":"Cristian","family":"Hob\u00e1n","sequence":"additional","affiliation":[]},{"given":"C\u00e9sar A.","family":"Murga-Moreno","sequence":"additional","affiliation":[]},{"given":"Jos\u00e9 M.","family":"Angulo-Tisoc","sequence":"additional","affiliation":[]},{"given":"Madoka","family":"Ichikawa-Seki","sequence":"additional","affiliation":[]}],"member":"297","published-online":{"date-parts":[[2022,10,20]]},"reference":[{"key":"5538_CR1","first-page":"40","volume":"2","author":"RM Kaplan","year":"2001","unstructured":"Kaplan RM. 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