Аннотация
Infection has been reported in humans and/or animals within 13 countries of the European Union (Pozio et al., 2013).The disease is endemic and represents a public health problem worldwide including Russian Federation, Ukraine, Belarus and Kazakhstan (Control of Food Borne Trematode Infections, 1995).Prevalence of opisthorchiasis in human reached a peak 2,521 cases in 2002, and gradually decreased to 1,225 cases in 2011 in Kazakhstan (Sultanov et al., 2014).In veterinary medicine, opisthorchiasis infection was often Research ArticleAbstract | Two species of opisthorchiid trematodes, Opisthorchis felineus (Rivolta, 1884) and Metorchis bilis (Braun, 1890), are known to infect humans and piscivorous mammals in Kazakhstan, causing opisthorchiasis or metorchiasis, respectively.There is difficulty in their differential diagnosis since their eggs are morphologically undistinguished and serological assays are low sensitive and specific.DNA sequenceing, especially for the nuclear ribosomal internal transcribed spacer 2 gene (ITS2), has already been used to differentiate between trematodes of the family Opisthorchiidae.The aim of this study is to establish a method for differentiation and identification of opisthorchiid species using molecular approach.Specific primers for a multiplex PCR were designed based on corresponding ITS2 gene sequence, and its detection limit was assessed by serial dilutions of the genomic DNAs of trematodes.Furthermore, samples of various parasites species of carnivores were tested using the developed assay.In this study, we developed species-specific primers for the differential detection of the two medically important trematodes based on the PCR assay.The speciesspecific primers were targeted to either variable or conserved regions of the ITS2 gene.ITS2 fragments with different sizes were amplified; 207 bp for O. felineus and 167 bp for M. bilis.Specificity was tested and no cross-reaction occurred and the sensitivity ranged between 2 and 1 ng of DNA template.The developed species-specific primers and PCR assay in this study could be applied for differential diagnosis of these medically and veterinary important opisthorchiid infections.