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Developing a Salivary Antibody Multiplex Immunoassay to Measure Human Exposure to Environmental Pathogens
Published on: September 12, 2016
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Statistical approaches to developing a multiplex immunoassay for determining human exposure to environmental
Swinburne A J Augustine1, Kaneatra J Simmons1, Tarsha N Eason2
1National Exposure Research Laboratory, U.S. Environmental Protection Agency, 26 W. Martin Luther King Drive, Cincinnati, OH 45268, USA.
Journal of Immunological Methods
|June 14, 2015
Summary
This study developed a multiplex immunoassay to detect multiple waterborne pathogens, improving upon costly single-analyte tests. The optimized assay accurately identified human exposure to various microbial contaminants in patient samples.
Area of Science:
- Environmental microbiology
- Immunology
- Biotechnology
Background:
- Waterborne pathogens pose significant public health risks, necessitating accurate exposure assessment.
- Conventional immunoassays (e.g., ELISAs) are limited by their single-analyte detection, high cost, and sample volume requirements.
- Multiplex immunoassays offer a solution for simultaneous detection of multiple pathogens, but require extensive optimization.
Purpose of the Study:
- To develop and optimize a multiplex immunoassay for detecting human exposure to waterborne pathogens.
- To address challenges in multiplex assay development, including antigen selection and reagent concentration optimization.
- To validate the assay's performance using diagnostically characterized patient samples.
Main Methods:
- A Design of Experiments (DOE) approach was employed to optimize reagent concentrations for antigen coupling to Luminex™ xMAP microspheres.
- Singleplex assay results for *Helicobacter pylori*, *Campylobacter jejuni*, *Escherichia coli* O157:H7, and *Salmonella typhimurium* informed multiplex assay parameters.
- Finite mixed modeling (FMM) was used to establish cut-offs for differentiating exposed and non-exposed individuals.
Main Results:
- The optimized multiplex immunoassay successfully detected Immunoglobulin G (IgG) antibodies against *H. pylori*, *C. jejuni*, *Toxoplasma gondii*, hepatitis A virus (HAV), rotavirus, and noroviruses.
- High detection rates were achieved: 87.5% for *H. pylori*, 100% for *T. gondii*, and 89% for HAV.
- The assay identified exposure to previously uncharacterized environmental pathogens in the tested plasma samples.
Conclusions:
- The developed multiplex immunoassay is a cost-effective and efficient tool for assessing human exposure to a range of waterborne pathogens.
- DOE and statistical modeling provide robust methods for optimizing and validating multiplex immunoassay performance.
- This approach enhances the ability to monitor and mitigate risks associated with microbial contaminants in water.
