A comparative Study between Blood Smear, Whole Blood PCR and FTA Card PCR for Diagnosis of Theileria annulata and Theileria orientalis in Cattle

Abstract

Bovine theileriosis, a tick borne haemoprotozoan disease, caused by, Theileria annulata and Theileria orientalis and responsible for heavy economical losses in dairy industry. Diagnosis of theileriosis is mainly based on clinical signs and conventional techniques. Polymerase chain reaction using whole blood is a modern method of diagnosis of theileriosis as it can detect infection even in low parasitemia but only concern is that its very time consuming and chances of loss of DNA is more. Hence, to overcome this, the present study reports the diagnosis of T. annulata and T. orientalis by comparing Whole blood PCR and FTA card PCR whereas a comparison between blood smear examination, whole blood PCR and FTA card PCR was done to determine the sensitivity of the techniques. The study was conducted on 50 blood samples of cattle having typical clinical signs and blood smear after staining with Giemsa’s stain were examined for presence of Theileria spp. in R.B.C. The primer sequences were used as per Kolte et al and Kakati et al. The assay employs primers specific for the gene encoding the 30-kDa major merozoite surface antigen of T. annulata and Major merozoite surface protein gene of T. orientalis. Out of total 50 blood samples, 16 were positive for Theileria sp. by blood smear examination, 19 were positive for T. annulata by Whole blood PCR and FTA card PCR whereas for T. orientalis, 26 were positive by whole blood and 27 samples by FTA card PCR. So the diagnosis of theileriosis by using FTA card PCR is found to be sensitive and less time consuming method as compared to Whole blood PCR.