Abstract
Waterborne infectious diseases are a major public health concern worldwide. Few methods have been established that are capable of measuring human exposure to multiple waterborne pathogens simultaneously using non-invasive samples such as saliva. Most current methods measure exposure to only one pathogen at a time, require large volumes of individual samples collected using invasive procedures, and are very labor intensive. In this article, we applied a multiplex bead-based immunoassay capable of measuring IgG antibody responses to six waterborne pathogens simultaneously in human saliva to estimate immunoprevalence in beachgoers at Boquerón Beach, Puerto Rico. Further, we present approaches for determining cutoff points to assess immunoprevalence to the pathogens in the assay. For the six pathogens studied, our results show that IgG antibodies against antigens from noroviruses GI.I and GII.4 were more prevalent (60 and 51.6%, respectively) than Helicobacter pylori (21.4%), hepatitis A virus (20.2%), Campylobacter jejuni (8.7%), and Toxoplasma gondii (8%) in the saliva of the study participants. The salivary antibody multiplex immunoassay can be used to examine immunoprevalence of specific pathogens in human populations.
Highlights
Acute gastrointestinal illness (AGI) has long been associated with swimming in fecally contaminated waters [1,2,3,4,5,6,7,8,9,10,11,12]
The Spearman rank-order correlation coefficient assessing the linear relationship between the median fluorescence intensity (MFI) of the antigens in the multiplex ranged from 0.20 for H. pylori/ norovirus GII.4 to 0.55 for C. jejuni/H. pylori (p-value < 0.05 for each pairing), indicating a statistically significant, weak to moderate positive correlation. This is the first multiplex immunoassay study aimed at describing the immunoprevalence of circulating antibodies to six waterborne pathogens in beachgoers in Puerto Rico
The results indicated that more than three-fifths of the beachgoers had evidence of prior exposure to noroviruses, while over a fifth were previously exposed to H. pylori and Hepatitis A virus (HAV)
Summary
Acute gastrointestinal illness (AGI) has long been associated with swimming in fecally contaminated waters [1,2,3,4,5,6,7,8,9,10,11,12]. Epidemiological surveys and enzyme-linked immunosorbent assays are often used to ascertain the cause of these illnesses. These approaches are time consuming, costly, and suffer from challenges such as selection bias and patient recollection of symptoms [13, 14]. The combination of saliva samples and a multiplex assay provides a powerful tool that could enhance health outcome assessment and measurement for certain types of epidemiological studies [17, 19, 35, 45, 46]
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