One hundred cattle-calves, out of which 37 cross bred and 63 indigenous, were screened for Theileria annulata infection during October-2015 to September-2016 brought to Teaching Veterinary Clinical Complex, College of Veterinary and Animal Science, Bikaner. Blood samples were collected in ethylene diamine tetra acetic acid (EDTA @1mg/ml) vacutainers from all the suspected cattle-calves for genomic DNA isolation and subjected for confirmatory diagnosis by nested polymerase chain reaction (nPCR). | Breed related hospital prevalence of Theileria annulata infection in cattle-calves confirmed by nested polymerase chain reaction in Bikaner district of Rajasthan India