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BurkPx: a multiplex serodiagnostic bead assay to detect Burkholderia pseudomallei exposures in non-human primates

Celona, Kimberly Rose (2021) BurkPx: a multiplex serodiagnostic bead assay to detect Burkholderia pseudomallei exposures in non-human primates. Masters thesis, Northern Arizona University.

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Melioidosis is a fatal zoonotic disease caused by the soil-dwelling Gram-negative bacteria Burkholderia pseudomallei. It is most well-known for its burden on humans, with an estimated 165,000 human cases per year worldwide, but this illness is equally important for animals, including non-human primates (NHPs). Although there is no published disease burden for NHPs we can look at the handful of reported melioidosis cases dating back to the 1920’s in multiple countries to make the conclusion that burden is not great, but possibly sporadic. Currently the gold standard for detecting B. pseudomallei in humans and animals is to culture from clinical specimens; however, the sensitivity of this technique is subpar. An alternative test, the indirect hemagglutination assay (IHA) is the standard serological assay for melioidosis, but also has low metrics. The search for a rapid, highly sensitive and specific assay is ongoing. Both direct detection methods like PCR and indirect detection methods like enzyme linked absorbent assays have been developed and evaluated. We have developed a multiplexed serological assay, called BurkPx, which uses purified B. pseudomallei proteins and carbohydrates bound to 21 fluorescently distinct MagPix magnetic beads to standardize a serodiagnostic assay for NHP. We screened B. pseudomallei-challenged (n=115) and non-challenged (n=126) rhesus macaques to explore time-dependent antibody responses and to train two multivariate models, LASSO and Ridge Regression, in differentiating exposure from non-exposure. When cross-validating the model using independent data, 59.6% sensitivity and 100% specificity was observed. However, removal of week 1 samples increased the sensitivity to 91.3%. To our knowledge, this is the first diagnostic tool, a serological multiplex assay, to be formally evaluated for NHPs and results in promising sensitivity and specificity metrics.

Item Type: Thesis (Masters)
Publisher’s Statement: © Copyright is held by the author. Digital access to this material is made possible by the Cline Library, Northern Arizona University. Further transmission, reproduction or presentation of protected items is prohibited except with permission of the author.
Keywords: diagnostic; Melioidosis; non-human primate; Burkholderia pseudomallei; Bacterial infections; Soil; Diagnosis
Subjects: R Medicine > RB Pathology
MeSH Subjects: C Diseases > C01 Bacterial Infections and Mycoses
NAU Depositing Author Academic Status: Student
Department/Unit: Graduate College > Theses and Dissertations
College of the Environment, Forestry, and Natural Sciences > Biological Sciences
Date Deposited: 23 Feb 2022 17:56
Last Modified: 23 Feb 2022 17:56
URI: https://openknowledge.nau.edu/id/eprint/5740

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