Epidemiological investigations of bovine brucellosis and evaluation of loop mediated isothermal amplification assay for field application
Muhammad Bilal1, Shafia Tehseen Gul1*, Muhammad Tariq Javed1, Muhammad Saqib2
1Department of Pathology, Faculty of Veterinary Sciences, University of Agriculture, Faisalabad, Pakistan
2Department of Clinical Medicine and Surgery, Faculty of Veterinary Sciences, University of Agriculture, Faisalabad, Pakistan
Abstract
Brucellosis is considered as a highly contagious and zoonotic disease globally and various diagnostic tests are available for its diagnosis. Keeping in view, the limitations of currently used serological techniques, a more precise, sensitive, and reliable loop mediated isothermal amplification (LAMP) assay was evaluated as an emerging diagnostic tool. In the current study, serum samples from cows (n=1989) and buffaloes (n=1467) were collected from the study area i.e., District Faisalabad and Toba Tek Singh in Punjab, Pakistan. As these two districts are present around the river Ravi in Punjab, Pakistan and known as the house of Nili Ravi breed. A number of dairy farms of local and imported cows are also present in this area. Initially the samples were screened by the RBPT and then subjected to c-ELISA for confirmation. Overall, 12.16 and 9.3% cows and buffaloes were seropositive through RBPT while 11.21 and 7.70% cows and buffaloes were seropositive via c-ELISA. The positive samples from c-ELISA were further subjected to molecular amplification at 1.5% agarose gel through LAMP assay. The current study concludes that LAMP assay is more sensitive as compared to other conventional PCR techniques while detecting true positives for brucellosis, so it can be used for confirmation of Brucella abortus as compared to PCR. In addition to the sensitivity and specificity and qualitative results can be observed through naked eye in LAMP, which is not possible in PCR.
Keywords: Brucellosis, LAMP, Quick diagnosis, Serological techniques, Rapid point of care
