Development of a Rapid Diagnostic Test for Antibiotic Resistance in Zoonotic Bacterial Pathogens

Project Abstract

The increasing prevalence of antibiotic-resistant zoonotic bacterial pathogens poses a significant threat to both veterinary and human health, necessitating the development of rapid and reliable diagnostic tools to detect resistance mechanisms efficiently. This study aims to develop a novel, cost-effective, and rapid diagnostic test that can accurately identify antibiotic resistance in key zoonotic bacterial pathogens such as *Salmonella spp.*, *Escherichia coli*, and *Campylobacter spp.*. The research combines molecular biology techniques, immunoassays, and biosensor technology to create a diagnostic platform that delivers results within a few hours, significantly reducing the turnaround time compared to traditional culture-based methods which often take days. The methodology involves the collection of bacterial isolates from clinical samples sourced from livestock, pets, and wildlife, followed by phenotypic antibiotic susceptibility testing using standardized disk diffusion and MIC methods to establish resistance profiles. Subsequently, genomic DNA extraction allows for the identification of resistance genes such as *bla*, *mecA*, *tet*, and *aad* through PCR amplification. These genetic markers guide the design of specific probes for the immunoassay component. An innovative biosensor-based detection system is developed by immobilizing these probes on a nanomaterial-enhanced platform, enabling high sensitivity and specificity in detecting resistance genes directly from samples without the need for extensive culturing. The test's performance is validated by comparing its results with conventional phenotypic and genotypic techniques across a diverse panel of clinical isolates. Sensitivity, specificity, accuracy, and predictive values are calculated to assess its diagnostic efficacy. Additionally, the study evaluates the test's cost-effectiveness and potential for field deployment in resource-limited settings where rapid decision-making is crucial to controlling zoonotic disease transmission. The results demonstrate that the developed diagnostic tool achieves over 95% concordance with standard methods, offering a rapid turnaround time of approximately 2-3 hours. It reliably detects multiple resistance genes simultaneously, providing comprehensive resistance profiles in a single test. This rapid diagnostic capability can facilitate timely interventions, antimicrobial stewardship, and policy formulation to curb the spread of resistant zoonotic bacteria. Overall, this research offers a significant advancement in veterinary microbiology by providing a practical solution for early detection of antibiotic resistance, which is vital for managing zoonotic infections effectively. The developed test's scalability and adaptability to various settings suggest its potential to become a standard diagnostic method in veterinary clinics, farms, and public health laboratories, contributing to global efforts in combating antimicrobial resistance.

Project Overview

This project is about creating a quick and easy test to find out if bacteria that can spread from animals to humans are resistant to antibiotics. These bacterial infections can be dangerous because if they do not respond to common medicines, they can cause serious health problems. The goal of the project is to help veterinarians and health workers quickly identify whether bacteria are resistant, so they can choose the right treatment faster and more effectively. The problem this project addresses is the increasing supply of bacteria that no longer respond to usual antibiotics, making infections harder to treat. When these bacteria come from animals, they can infect people as well, leading to a public health risk. Traditional tests to detect antibiotic resistance are often slow, taking several days, which delays treatment and can worsen health outcomes. The researcher will first review existing methods of testing antibiotic resistance in bacteria from animals. Then, they will focus on developing a new, faster test, possibly using special dyes or markers that change color when bacteria are resistant. To do this, they will collect samples from infected animals, isolate bacteria, and test their resistance using both standard methods and the new rapid test. The researcher will compare the results to see if the new test is accurate and quicker than current options. Next, the project may involve refining the test to make it reliable, affordable, and suitable for use in veterinary clinics or even in rural areas. The expected outcome is a simplified test that provides results within a few hours instead of days, so veterinarians can treat animals more effectively and help prevent the spread of resistant bacteria. Ultimately, this project aims to improve animal health, protect human health, and combat antibiotic resistance more efficiently by providing a practical tool for early detection.