Cloning, expression and characterization of the taenia multiceps HSP70 gene and investigation of the test kit properties

Taenia multiceps is a taeniid cestode that lives in the small intestine of adult canines and other carnivores. Its larva is known as Coenurus cerebralis and is located in the nervous system of sheep and other ruminants, including the brain and medulla spinalis. The presence of the cyst causes typical neurological symptoms and most cases result in the death of the animals. Coenurosis can cause significant economic losses to sheep farms as it commonly affects young animals. The aim of this thesis was to determine the efficacy of the recombinant protein expressed by T. multiceps HSP70 gene in the diagnosis of sheep coenurosis in sheep and to develop a kit applicable in field conditions. For this purpose, primers were designed to amplify the gene encoding HSP70 from the whole genome sequence of T. multiceps, mRNA was isolated from C. cerebralis cyst obtained from an infected sheep's brain and then cDNA was isolated. The gene region related to the designed primers was amplified by PCR and the product obtained was cloned and expressed. The efficiency of the recombinant protein purified afterwards was demonstrated by Western blot using known positive and negative sheep sera. In addition, the efficiency of the recombinant protein in ELISA was determined with sera collected from 1207 sheep. As a result of these procedures, the sensitivity and specificity values of the recombinant TmHSP70 antigen in the western blot test were 100%, while the sensitivity and specificity rates of the ELISA test were 66.6% and 52.6%, respectively. At the same time, 567 (46.97%) of the 1207 serum samples analyzed were found to be positive by ELISA. As a result, it has been shown that recombinant TmHSP70 antigen can be used in a western blot-based test for the diagnosis of coenurosis in sheep.