Development of Point-of-Care Diagnostic Tool for Human Brucellosis - SURF Spring 2021

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  • May 6, 2021
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  • Iyamu, Osahon
    • Affiliation: College of Arts and Sciences, Department of Psychology and Neuroscience
    • Other Affiliation: Department of Chemistry
Abstract
  • Brucellosis is a zoonotic disease caused by the gram-negative bacteria of the genus Brucella. This infectious disease has a tremendous economic and social impact in regions in central Africa. An estimated $500 million to $1 billion USD income potential could be generated in central Africa if the disease is eradicated. Brucellosis infections can cause symptoms such as spontaneous abortions, chronic infection, coughing, muscle pain, fevers, and many other peripheral organ complications. Despite these pervasive issues caused by the disease, challenges remain for its diagnosis. Not only is it difficult to differentiate the symptoms from other infectious diseases like malaria, but common means for diagnosing Brucellosis are also both impractical and nonspecific. These concerns necessitate the creation of a new cost-effective, sensitive, specific, and user-friendly approach to diagnosing Brucellosis. Recombinase Polymerase Amplification (RPA) is an isothermal DNA amplification technique that utilizes single-stranded binding proteins (SSBs), primers, and oligonucleotides to generate amplified DNA in under 30 minutes. RPA used in conjunction with lateral flow (LF) platforms creates an effective means for detecting nucleic acids in comparison to other methods. In this study, we designed and optimized a Brucella RPA-LF assay to target the most common bacterial strains observed in human Brucellosis. The assay appears to have a lower limit of detection of 1.37 genomes by probit analysis and demonstrates high specificity for Brucella strains. Future directions will look at other methods for extracting DNA necessary to run the RPA-LF assay and implementing the diagnostic test in southwestern rural Uganda to clinically validate the assay.
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