Encephalitozoon spp. u volně žijících drobných savců – identifikace pomocí nested PCR a genotypizace
| Title in English | Encephalitozoon spp. in wild-type mammals - identification by nested PCR and genotyping |
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| Authors | |
| Year of publication | 2018 |
| Type | Conference abstract |
| MU Faculty or unit | |
| Citation | |
| Description | 1339/5000 In addition to basic single PCR, there are several modifications, one of which is nested PCR, which consists of two successive reactions. The aim of my work was to compare the sensitivity of these two methods for the detection of parasites Encephalitozoon spp. and at the same time to find out the spread of this parasite in wild small mammals on the territory of the Czech Republic. To isolate DNA, 100 samples of small rodents and insectivores were collected in the areas of the Protected Landscape Area of the Moravian Karst and Poodří Protected Landscape Area. In the case of single PCR, 2 primers specific for E. cuniculi were used, this parasite was detected in one individual (1 percent). Two sets of primers used in nested PCR allow detection of multiple microsporidial parasites. The distinction of individual genera is Poodří Protected Landscape Area (14.9 percent), of the individuals captured in the Moravian Karst Protected Landscape Area were positive 5.7 percent of the animals. To distinguish individual species, positive samples were sent for sequencing to Macrogen's laboratories. Genotyping was successfully performed on only three samples, based on the comparison of the obtained sequences with the data reported in the BLAST database, in all three cases it was E. cuniculi. Based on the result, the nested PCR is against single PCR for the detection of parasites Encephalitozoon spp. significantly more sensitive and due to the size of the resulting PCR product. Parasites of Encephalitozoon were detected in 10 individuals (10 percent). |