Characterization of enteric disease in children using a low-cost specimen preservation method.

BACKGROUND 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 filter paper specimen preservation can identify diarrheal diseases causing significant morbidity among children in resource constrained countries. METHODS A sub-study was nested into cholera surveillance in Cameroon. Enrollment criteria included: enrollment between 8/1/16 - 10/1/18; age < 18 years; a stool specimen; ≥ three loose stools within 24hours with the presence of dehydration and/or blood. 7227 persons were enrolled, for which 2746 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. RESULTS 107 were ETEC positive of which: 40.2% (N=43) LT-STh, 19.6% (N=21) LT-STp; and 49.5% (53) LT-only. Major CFs were present in 43.9% of ETEC-positives. 96 were positive for Shigella, of which 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. Shigellae and rotavirus were most strongly associated with diarrhea in children with mixed infections. CONCLUSION 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.