Population structure, molecular characterization, and phylogenetic analysis of Fasciola gigantica from two locations in Uganda

Vudriko, Patrick; Echodu, Richard; Tashiro, Michiyo; Oka, Nozomi; Ichikawa‐Seki, Madoka

doi:10.1016/j.meegid.2022.105359

Cited by 1 publication

(1 citation statement)

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“…A total of 1312 Fasciola flukes (470 F. hepatica, 609 F. gigantica, and 233 hybrid Fasciola) from 11 countries (Afghanistan, Algeria, Peru, Spain, Indonesia, Malaysia, Nigeria, Pakistan, Uganda, Japan, and Bangladesh) [8,9,11,12,[14][15][16][17][18][19][20] were used in the present study. Fragment analyses of nuclear pepck and pold and the nucleotide sequencing of mitochondrial nad1 have been performed in previous studies [8,9,11,12,[14][15][16][17][18][19][20]. Discrepancies between pepck and pold were observed among 7, 19, 6, 27, and 15 Fasciola isolates from Afghanistan, Algeria, Peru, Spain, and Nigeria, respectively.…”

Section: Fasciola Samplesmentioning

confidence: 99%

Development of novel DNA marker for species discrimination of Fasciola flukes based on the fatty acid binding protein type I gene

et al. 2022

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Background Multiplex polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) for nuclear phosphoenolpyruvate carboxykinase (pepck) and polymerase delta (pold), respectively, have been used to differentiate Fasciola hepatica, F. gigantica, and hybrid Fasciola flukes. 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 (FABP) type I gene. Methods Nucleotide sequence variations of FABP type I were analyzed using DNA samples of F. hepatica, F. gigantica, and hybrid Fasciola flukes obtained from 11 countries in Europe, Latin America, Africa, and Asia. A common forward primer for F. hepatica and F. gigantica and two specific reverse primers for F. hepatica and F. gigantica were designed for multiplex PCR. Results Specific fragments of F. hepatica (290 bp) and F. gigantica (190 bp) were successfully amplified using multiplex PCR. However, the hybrid flukes contained fragments of both species. The multiplex PCR for FABP type I could precisely discriminate the 1312 Fasciola samples used in this study. Notably, no discrimination errors were observed with this novel method. Conclusions Multiplex PCR for FABP type I can be used as a species discrimination marker in place of pepck and pold. The robustness of the species-specific primer should be continuously examined using a larger number of Fasciola flukes worldwide in the future since nucleotide substitutions in the primer regions may cause amplification errors. Graphical abstract

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