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Published on: March 10, 2020
Characterization of Enteric Disease in Children by Use of a Low-Cost Specimen Preservation Method
Amanda K Debes1, Shaoming Xiao1, Jie Liu2
1Department of International Health, Johns Hopkins Universitygrid.21107.35 Bloomberg School of Public Health, Baltimore, Maryland, USA.
Insights
Dried filter paper preserves stool specimens for identifying pediatric diarrhea pathogens like rotavirus, Shigella, and ETEC. This method simplifies surveillance in resource-limited settings, aiding vaccine development.
Area of Science:
- Infectious Diseases
- Molecular Diagnostics
- Public Health Surveillance
Background:
- Diarrhea is a primary cause of mortality in children under five globally.
- Current molecular diagnostic methods for enteric pathogens require frozen stool specimens, posing logistical challenges in resource-constrained settings.
- Dried stool specimens on filter paper offer a viable alternative for pathogen detection.
Purpose of the Study:
- To evaluate the efficacy of dried filter paper-preserved stool specimens for identifying enteric pathogens causing significant diarrheal disease in children.
- To assess the burden of specific enteric pathogens, including enterotoxigenic Escherichia coli (ETEC), Shigella, and rotavirus, in pediatric diarrhea cases.
- To demonstrate the utility of this preservation method for enteric disease surveillance in resource-limited areas.
Main Methods:
- A substudy nested within cholera surveillance in Cameroon enrolled children (<18 years) with acute diarrhea between August 2016 and October 2018.
- Stool specimens were preserved on filter paper and analyzed using the enteric TaqMan array card.
- Pathogen detection was correlated with clinical data, including diarrhea severity, dehydration, and presence of blood.
Main Results:
- Enterotoxigenic E. coli (ETEC) was detected in 107 children, with various toxin profiles (LT, STh, STp).
- Shigella was identified in 96 children, with 14.6% reporting dysentery.
- Rotavirus, Shigella, and ETEC were confirmed as major contributors to pediatric diarrhea, with Shigella and rotavirus strongly associated with severe cases and mixed infections.
Conclusions:
- Dried filter paper-preserved stool specimens effectively identify key pediatric diarrhea pathogens, eliminating the need for cold chain storage.
- This method significantly enhances the feasibility of enteric surveillance in resource-limited countries.
- Findings support the use of this technique to better understand diarrheal disease burden and guide vaccine development and implementation strategies.
Abstract:
Diarrhea is a leading cause of death in children under five. Molecular methods exist for the rapid detection of enteric pathogens; however, the logistical costs of storing stool specimens limit applicability. We sought to demonstrate that dried specimens preserved using filter paper can be used to identify diarrheal diseases causing significant morbidity among children in resource-constrained countries. A substudy was nested into cholera surveillance in Cameroon. Enrollment criteria included enrollment between 1 August 2016 and 1 October 2018, age of <18 years, availability of a stool specimen, and having three or more loose stools within 24 h with the presence of dehydration and/or blood. A total of 7,227 persons were enrolled, of whom 2,746 met enrollment criteria and 337 were included in this analysis using the enteric TaqMan array card. Bacterial pathogens were compared to severity of diarrhea, age, and sex, among other variables. One hundred seven were positive for enterotoxigenic Escherichia coli, of which 40.2% (n = 43) had heat-labile enterotoxin (LT) and the heat-stable enterotoxin STh, 19.6% (n = 21) had LT and the heat-stable enterotoxin STp, and 49.5% (n = 53) had LT only. Major colonization factors (CFs) were present in 43.9% of enterotoxigenic E. coli (ETEC)-positive patients. Ninety-six were positive for Shigella, of whom 14 (14.6%) reported dysentery. Model-derived quantitative cutoffs identified 116 (34.4%) with one highly diarrhea-associated pathogen and 16 (4.7%) with two or more. Shigella and rotavirus were most strongly associated with diarrhea in children with mixed infections. Dried-filter-paper-preserved specimens eliminate the need for frozen stool specimens and will facilitate enteric surveillance and contribute to the understanding of disease burden, which is needed to guide vaccine development and introduction. This study confirms rotavirus, Shigella, and ETEC as major contributors to pediatric diarrheal disease in two regions of Cameroon.
