DEVELOPMENT OF DRY LAMP AS POINT OF CARE DIAGNOSTICS FOR THE DIAGNOSIS OF BOVINE THEILERIOSIS
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Date
2022
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KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR - 585 401
Abstract
Tropical theilerosis is an economically important vector-borne disease of tropical and subtropical parts of the world including India. Though there are many advancements being made for the diagnosis of theileriosis, they could not be efficiently applied to the field level. Hence, a study was undertaken to develop a dry LAMP assay as point of care diagnostics of diagnosis of theileriosis in cattle. Both wet and dry LAMP assays targeting cytochrome b gene were standardized for diagnosis of Theileria annulata in cattle and comparative evaluation was done. To state in terms of sensitivity it was found that both wet LAMP and dry LAMP could amplify as least as 10-4ng (0.1pg) of DNA. In terms of specificity it was found that both wet and dry LAMP assay amplified only Theileria annulata and did not amplify other closely related haemoprotozoans. A total of 200 suspected blood samples were subjected to microscopy, PCR , wet LAMP and dry LAMP assay. It was found that 16 samples were found to be positive for theilerosis in microscopy, 20 samples were found to be positive in PCR and 23 samples were found to be positive in both wet and dry LAMP assay. Hence, we can conclude that the LAMP assay (dry and wet LAMP) was found to be more sensitive (100%) when compared to microscopy (69.5%) and PCR (86.9%). In terms of shelf life, it was observed that the dry LAMP reaction tubes at room temperature as well as refrigeration temperature provided successful amplification till 7 weeks. Hence it can be concluded that using dry LAMP assay, one can develop a point of care diagnostic kit for the field level diagnosis of T.annulata in cattle.