Development of a recombinant TaSP-based Dot-ELISA for detection of Theileria annulata infection in cattle

• Published in: Ticks and tick-borne diseases Vol. 9; no. 6; pp. 1416 - 1420
• Main Authors: Mohmad, Aquil, Chandra, D., Saravanan, B.C., H.V, Manjunathchar, O.R, Vinodh Kumar, Fular, Ashutosh, Chigure, Gajanan, Kaur, Navneet, Ghosh, S.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier GmbH 01-09-2018
• Subjects: Animals; Antibodies, Protozoan - blood; Antigens, Protozoan - immunology; Cattle; Dot-ELISA; ELISA; Enzyme-Linked Immunosorbent Assay - methods; Enzyme-Linked Immunosorbent Assay - veterinary; Field infection; Protozoan Proteins - immunology; r-TaSP; Recombinant Proteins - immunology; Theileria annulata; Theileria annulata - isolation & purification; Theileriasis - diagnosis; Theileriasis - parasitology; Vaccines, Synthetic - therapeutic use; Dot-ELISA; Field infection; Cattle; Theileria annulata; ELISA; r-TaSP
• ISSN: 1877-959X; 1877-9603
• DOI: 10.1016/j.ttbdis.2018.06.016

The study was conducted to develop and validate Dot-ELISA for the diagnosis of Theileria annulata infection in cattle using recombinant Theileria annulata surface protein (r-TaSP). The r-TaSP based indirect plate-ELISA was used as a reference test to compare the efficacy of the Dot-ELISA. The Dot-ELISA was optimized with 500 ng of antigen per dot, 1:150 dilution of serum and 1:1000 dilution of secondary antibody for positive and negative reaction. A total of 17 confirmed positive, 25 negative and 129 field sera samples were used to calculate the diagnostic accuracy of Dot-ELISA in comparison with indirect plate-ELISA. The diagnostic sensitivity and specificity of the Dot-ELISA was 95.8 per cent (95% CI, 93.1–97.2) and 80 per cent (95% CI, 48.1–96.2), respectively. The positive predictive value (PPV) of Dot-ELISA was 98.2 percent (95% CI, 95.5–99.7) and negative predictive value (NPV) was 61.6 percent (95% CI, 37–74). The positive and negative likelihood ratios were 4.79 (95% CI, 1.8–25.69) and 0.053 (95% CI 0.03–1.4), respectively. The Dot-ELISA showed moderate agreement (k value, 0.67, 95% CI, 0.36– 0.82) with indirect plate-ELISA. The developed Dot-ELISA is less expensive and convenient for the diagnosis of T. annulata infection in cattle under field conditions.