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Statistical approaches to developing a multiplex immunoassay for determining human exposure to environmental pathogens.
Citation:
Augustine, S., K. Simmons, T. Eason, S. Griffin, C. Curioso, L. Wymer, Shay Fout, A. Grimm, K. Oshima, AND A. Dufour. Statistical approaches to developing a multiplex immunoassay for determining human exposure to environmental pathogens. JOURNAL OF IMMUNOLOGICAL METHODS. Elsevier Science Ltd, New York, NY, 425(10):1-9, (2015).
Impact/Purpose:
This method provides a novel tool that can be used to study human exposure to environmental pathogens, particularly waterborne pathogen exposure and swimming-associated illnesses, using a rapid, cost-effective, multiplex approach using human saliva. We present several approaches to analyze multiplex immunoassays results to measure immunoprevalence and immunoconversions (incident infections. Additionally, this method will provide a path forward to measure co-infections. When used in conjunction with survey type epidemiology studies, this method provides a powerful tool that can be used to measure asymptomatic infections that were previously difficult to measure.
Description:
This paper describes the application and method performance parameters of a Luminex xMAP™ bead-based, multiplex immunoassay for measuring specific antibody responses in saliva samples (n=5438) to antigens of six common waterborne pathogens (Campylobacter jejuni, Helicobacter pylori, Toxoplasma gondii, hepatitis A virus (HAV) and noroviruses (GI.1 and GII.4 genotypes). Three samples were collected over a six-week period, and analyzed using a multiplexed immunoassay to assess prior exposures (immunoprevalence in the initial sample) and immunoconversions, which serve as indicators of incident infections, to one or more of these pathogens. Methods were evaluated to establish cut-off points based on a control no-antigen bead set. These cut-offs were also used to develop criteria for estimating immunoconversions. The analysis of baseline (S1) saliva samples (n=2078) revealed the following average immunoprevalence rates based on all of the approaches used: noroviruses (GI.1: 64%, GII.4: 56%), H. pylori (26%), HAV (23%), T. gondii (13%) and C. jejuni (13%). For immunoconversions in individuals who provided all three prospectively collected samples (n=1399), we found that the overall immunoconversion rate ranged from about 5% to 15% depending on the criteria used. This work demonstrates that the combination of saliva collection and Luminex xMAP™ technology may provide a cost-effective and practical approach to identifying previous exposure and infection patterns in large populations.
