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    Absolute Quantitative PCR Detection of Shigella spp./EIEC and Association with Moderate to Severe Diarrheal Illness in Children in Developing Countries

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    Author
    Lindsay, Brianna R.
    Advisor
    Stine, Oscar Colin
    Date
    2014
    Type
    dissertation
    
    Metadata
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    Abstract
    Background: Shigella spp. and enteroinvasive Escherichia coli (EIEC) are prominent enteric pathogens and estimates of prevalence are limited by suboptimal sensitivity of current detection methods. Recent studies have suggested common co-infection and frequent asymptomatic pathogen detection. Objective: The objective of this research is to examine the association between the quantity of Shigella/EIEC identified in stool and moderate-to-severe diarrhea (MSD) and how this association is modified by different factors such as age, location, co-infecting pathogens and the gut microbiota. Methods: The Global Enteric Multicenter Study (GEMS) estimated the etiology and population-based burden of MSD in sub-Saharan Africa and South Asia in children under the age of five years. Here, we conducted a case-control study sampled from GEMS utilizing quantitative polymerase chain reaction (QPCR) for the identification of the Shigella/EIEC ipaH gene in stool samples and compared the results of conventional culture to those of QPCR. Using multivariate logistic regression modeling we tested for effect modification and interaction and examined the effects of co-infecting pathogens identified through traditional microbiologic techniques and 16s rRNA sequencing. Results: A cut-point of 1.4x104 ipaH gene copies results in an increased burden of MSD cases attributable to Shigella/EIEC. Both site and age were identified as significant effect modifiers of the relationship between ipaH quantity and MSD, with the greatest association seen in children 24-59 months of age (Odds Ratio 8.2; 95% CI 4.3-15.7) and from Bangladesh (OR13.2; 95% CI 7.3-23.8). After adjusting for age and site, rotavirus exhibited a significant negative association with high ipaH quantities in cases (OR=0.31, 95% CI 0.17-0.55) and resulted in large combined effects (OR=28.85; 95% CI 3.77-220). Five Lactobacillus taxa exhibited significant additive antagonistic effects, decreasing the association between Shigella and MSD, consistent with a possible protective role. Conclusions: QPCR detection of the ipaH gene substantially increases the fraction of MSD attributable to Shigella/EIEC above that attributed by culture and identifies similar age and site effects. Our results suggest that Lactobacillus may specifically inhibit the pathogenicity of Shigella/EIEC and conclude that future studies should continue to consider the effects of significant characteristics such as age and site as well as co-occurring species.
    Description
    University of Maryland, Baltimore. Epidemiology and Preventive Medicine. Ph.D. 2014
    Keyword
    diagnostics
    Diarrhea
    Epidemiology
    Microbiota
    Shigella
    Identifier to cite or link to this item
    http://hdl.handle.net/10713/4075
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    Theses and Dissertations All Schools
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