Multiplex PCR and Sequence Analysis to Investigate Genetic Diversity of Isolates from Cattle and Sheep in Turkey.

Fasciolosis is a highly prevalent helminthic infection caused by and . With the aim of identifying hybrid flukes, multiplex PCR was performed to amplify the gene. Furthermore, to determine haplotypes, mitochondrial gene was amplified and sequenced, and phylogenetic analyses were performed. Adult isolates were collected from 51 cattle and 51 sheep, genomic DNA was isolated, and one-step multiplex PCR was subsequently performed to amplify . Isolates that generated a 510 bp band were identified as , those that generated a 241 bp band were identified as , and those that generated both bands were identified as hybrid (aspermic) flukes. Multiplex PCR data identified four isolates as and 84 as . Fourteen hybrid isolates (five cattle and nine sheep) were identified. On unidirectional DNA sequence analysis of PCR products, three sequences were identified as and 99 as . In addition, only 4 of 87 haplotypes detected for sequences were found to be previously reported, while the remaining 83 are unique to this study. To summarize, this study is the first to report the existence of hybrid flukes and 83 unique haplotypes of in Turkey.