PRECISION IN PATHOGEN DETECTION: ENHANCED LAMP ASSAY FOR BRUCELLA ABORTUS WITH AND WITHOUT LOOP PRIMERS

Abstract

Brucellosis, a zoonotic disease caused by Brucella species, is a significant public health concern worldwide. The Loop-Mediated Isothermal Amplification (LAMP) assay is a rapid and sensitive diagnostic tool that can be used to detect Brucella abortus. In the current study, we evaluate the efficiency of LAMP assay with and without loop primers with a set of primers used to detect Brucella abortus. We found that the addition of loop primers did not enhance the amplification or detection capacity of the LAMP assay. All samples were successfully amplified in this study without the use of loop primers and hence the results indicate that loop primers are not always necessary when carrying out the LAMP assay for identification of Brucella abortus. It is different from earlier investigations that indicated optimum configuration of LAMP assays with the incorporation of loop primers. Because of these findings, it is possible to conclude that the position of loop primers applied in LAMP assays may depend on the target sequence and, thus, the possibility of their exclusion can be considered as the best strategy. The findings of this particular study have implications to the design and enhancement of the LAMP assays for the identification of Brucella abortus and also other diseases. This allows for the reduction in the number of primers used in LAMP assays, contributing to lowering the overall cost and the level of complexity associated with this technique, which is important for point-of-care diagnostics in regions of varying resource availability.