Design of Theileria spp. specific primers and DNA amplification protocols for molecular characterization

Abstract

Climate change is happening in much faster and stronger way than expected. One of global climates change most discussed consequences is its effect on infectious diseases. Even though it is not clear how infectious diseases will spread, or how their occurrence will change, current assumption is that, climate conditions are what limit the distribution of infectious diseases. This assumption is particularly true for tick – borne diseases, where relationship between vectors that spread disease and climate they are living in is well defined. Therefore, tick – borne diseases spread is going to change with the change of the climate. Theileria genus consists of intracellular protozoan that cause infectious disease in its host and are transmitted by ticks. The pathogen not only has complex life cycle comprising of both, the host and vector, but also has few individual species that has no cure of now and cause great loss in industry yearly. A lot is known of Theileria species that are distributed in tropical regions, but there is lack of information on species spread in Europe. There is almost no information on vectors that are responsible of distributing Theileria species in Europe. Because of that and unavoidable consequences of climate change Theileria species that are spread in Europe have to be very well observed and investigated. There is a need of a highly specific and sensitive method specific for Theileria detection. Using PCR and primers specific for Theileria genus, there is a possibility to define not only animals that are carrying the pathogen but also its vectors. One of possible genes for creating molecular primers to catch all species within genus of Theileria is 18S rRNR. In this study we look at V4 variable region of 18S rRNA gene for possible molecular primers, design them and optimize PCR for Theileria spp. detection