| Autor: |
Mishra PKK; Unit of Teaching Veterinary Clinical Complex, Faculty of Veterinary and Animal Sciences, Institute of Agricultural Sciences, RGSC, Banaras Hindu University, Mirzapur 231001, Uttar Pradesh, India., Jena A; Fisheries and Animal Resource Development Department, Bhubaneswar 751001, Odisha, India., Sarkhel SP; Department of Veterinary Parasitology, Faculty of Veterinary and Animal Sciences, Institute of Agricultural Sciences, RGSC, Banaras Hindu University, Mirzapur 231001, Uttar Pradesh, India., Behera SK; Department of Epidemiology & Public Health, Central University of Tamil Nadu, Tiruvarur 610001, Tamil Nadu, India., Das A; Fisheries and Animal Resource Development Department, Bhubaneswar 751001, Odisha, India., Sabarinath T; Clinical Bacteriological Laboratory, Indian Council of Agricultural Research-Indian Veterinary Research Institute, Nainital 263138, Uttarakhand, India., Jena D; Unit of Teaching Veterinary Clinical Complex, Faculty of Veterinary and Animal Sciences, Institute of Agricultural Sciences, RGSC, Banaras Hindu University, Mirzapur 231001, Uttar Pradesh, India., Mandal KD; Unit of Teaching Veterinary Clinical Complex, Faculty of Veterinary and Animal Sciences, Institute of Agricultural Sciences, RGSC, Banaras Hindu University, Mirzapur 231001, Uttar Pradesh, India., Sahu A; Department of Veterinary Parasitology, College of Veterinary Science and Animal Husbandry, Odisha University of Agriculture and Technology, Bhubaneswar 751003, Odisha, India., Kumar A; Department of Animal Genetics and Breeding, Faculty of Veterinary and Animal Sciences, Institute of Agricultural Sciences, RGSC, Banaras Hindu University, Mirzapur 231001, Uttar Pradesh, India., Kumar V; Unit of Teaching Veterinary Clinical Complex, Faculty of Veterinary and Animal Sciences, Institute of Agricultural Sciences, RGSC, Banaras Hindu University, Mirzapur 231001, Uttar Pradesh, India., Kadam RG; Unit of Teaching Veterinary Clinical Complex, Faculty of Veterinary and Animal Sciences, Institute of Agricultural Sciences, RGSC, Banaras Hindu University, Mirzapur 231001, Uttar Pradesh, India., Sathapathy S; Department of Veterinary Anatomy and Histology, College of Veterinary Science and Animal Husbandry, Odisha University of Agriculture and Technology, Bhubaneswar 751003, Odisha, India., Rao TP; Department of Veterinary Physiology and Biochemistry, Sri Venkateswara Veterinary University, College of Veterinary Science, Garividi 535101, Andhra Pradesh, India. |
| Abstrakt: |
Tick-borne diseases (TBDs) of livestock are endemic across various parts of tropical countries. Theileriosis is one such economically important TBD, caused by the Theileriidae family of organisms, which is transmitted by ticks. Theileria annulata , the causative agent of tropical theileriosis, contributes a significant loss to the dairy sector by causing anorexia, high fever, anemia, inflammatory changes in vital organs and icterus, thus, a loss in milk yield. Though vaccines are available, their protective efficacy is not absolute, and treatment is limited to early diagnosis of the causative agent. Routinely, microscopic identification of piroplasms in the erythrocytes (Giemsa-stained) of infected animals or schizonts in lymph node biopsies are practiced for diagnosis. PCR-based techniques (multiplex, uniplex, nested and real-time) have been reported to perform well in diagnosing active infection. Several attempts have been made using serological assays like Dot blot, ELISA and ICT, but the results were of variable sensitivity and specificity. Recombinant proteins like the Theileria annulata merozoite surface antigen (Tams1) and Theileria annulata surface protein (TaSP) have been explored as antigenic candidates for these assays. In the present study, we predicted an immunogenic peptide, i.e., TaSP-34, from the TaSP using various computational tools. The predicted peptide was custom synthesized. The diagnostic potential of the peptide was assessed by indirect plate ELISA to detect the bovine-IgM against Theileria annulata . Alongside, a recombinant truncated TaSP (rTaSP(tr)) was expressed and purified, which was used to compare the performance of the peptide as a diagnostic candidate. The IgM-based peptide ELISA was 100% sensitive and 92.77% specific as compared to PCR (Tams1 targeting), while 98.04% sensitivity and 97.44% specificity were observed in comparison with rTaSP(tr) ELISA. Almost perfect agreement between peptide ELISA and Tams1 PCR was observed with a Cohen's kappa coefficient (κ-value) of 0.901 and agreement of 95.31%. Further, the κ-value between the peptide ELISA and rTaSP(tr) ELISA was found to be 0.95, and the agreement was 97.65%, which shows a good correlation between the two tests. The findings suggest that the TaSP-34 peptide can be an efficient and new-generation diagnostic candidate for the diagnosis of T. annulata. Furthermore, the peptide can be synthesized commercially at a larger scale and can be a cost-effective alternative for the protein-based diagnostic candidates for T. annulata. |
