Using the fatty acid binding protein type I gene as a unique DNA marker, Fasciola flukes can be differentiated by species.
Abstract Background Fasciola hepatica, F. gigantica, and hybrid Fasciola flukes have been distinguished using multiplex polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) for nucleus phosphoenolpyruvate carboxykinase (pepck) and polymerase delta (pold), respectively. However, both approaches have been reported to exhibit discrimination flaws. The objective of this study was to create a multiplex PCR based on the FABP type I gene, a novel nuclear marker. Methods Using DNA samples of hybrid Fasciola flukes, F. hepatica, and F. gigantica collected from 11 countries in Europe, Latin America, Africa, and Asia, nucleotide sequence variants of FABP type I were examined. For multiplex PCR, two distinct reverse primers for F. hepatica and F. gigantica as well as a common forward primer were created. Results Using multiplex PCR, specific segments of F. hepatica (290 bp) and F. gigantica (190 bp) were amplified satisfactorily. The hybrid flukes, however,...