Vol. 12, Special Issue 11 (2023)

Author(s):
Sanjana, Marcia Ashmi, Sonu S Nair, Hilari Debbarma, Moon Moon Satpathy, Ambika Arun, Deepak Kumar, Abhishek, Ravi Kant Agrawal and Bablu Kumar

Abstract:
Brucellosis is a concern in many regions, especially developing countries. The methods for detecting diseases based on nucleic acids, like LAMP, provide a rapid and user-friendly approach to disease identification. These approaches hold the promise of expediting early disease detection and significantly reducing disease transmission. In the current investigation, an Isothermal LAMP test was developed and validated as a specific and rapid method for Brucella spp. detection in clinical samples. The primers designed and custom synthesized, were tailored to target the genus-specific BruAb2_0672 gene sequence of Brucella. The detection limit of BruAb2_0672 LAMP was 28 fg of Brucella genomic DNA, whereas conventional PCR detected 2.8 pg of template extracted from pure culture. Out of 10 blood samples from cattle spiked with Brucella abortus S19, nine samples tested positive with both PCR and RT-PCR when using the BruAb2_0672 LAMP outer primers. Additionally, all ten samples tested positive using LAMP. Furthermore, all ten negative samples were confirmed negative when subjected to PCR, RT-PCR, and LAMP.