Development of rapid immunological and molecular tools for diagnosis of brucellosis infection in indian population

Abstract

newline Abstract : newlineBackground: Brucellosis is a globally prevalent zoonotic disease that poses significant diagnostic challenges due to its nonspecific clinical manifestations and limitations in existing diagnostic tools. Conventional diagnostic approaches such as culture and serology often suffer from long turnaround times, low sensitivity, and risks of laboratory-acquired infections. There is an urgent need for rapid, accurate, and field-deployable diagnostic methods, particularly in resource-limited and endemic settings. newlineAim and Objectives: newlineThis study aimed to develop and evaluate novel molecular and immunological assays for the rapid and accurate diagnosis of Brucellosis. The specific objectives were: newline1. To develop a LAMP-PCR assay that targets Bcsp31 and VirB genes for the rapid and accurate molecular diagnosis of human brucellosis, enabling point-of-care testing with reduced turnaround time. newline2. To evaluate the performance of the developed LAMP-PCR assay in pilot samples, assessing its sensitivity, specificity, and accuracy, and comparing its results with conventional diagnostic methods. newline3. To develop antigenic peptide-based ELISA and whole-cell lysate-based ELISA assays using Brucella abortus and Brucella melitensis strains for enhanced immunological diagnosis of brucellosis, ensuring high sensitivity and specificity. newline4. To evaluate the performance of the antigenic peptide-based and whole-cell lysate-based ELISA assays in confirmed and suspected brucellosis cases, comparing the results with existing IgM commercial ELISA assays to determine the diagnostic specificity, concordance and diagnostic reliability. newlineMethods: A diagnostic laboratory-based study was conducted with a sample size of 312 participants. Molecular detection was carried out using both conventional PCR and the newly developed LAMP PCR assays. For immunological detection, two ELISA kits were developed: one based on antigenic peptides (Omp25, Omp31, LPS, VirB) and the other on Brucella WCL. Assay newlineperformances were evaluated against the ref