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Identification of Enteropathogens by Multiplex PCR among Rural and Urban Guatemalan Children with Acute Diarrhea
James T Gaensbauer1,2,3, Molly Lamb2, Diva M Calvimontes4
1Department of Pediatrics, Denver Health Hospital Authority, Denver, Colorado.
Insights
Multiplex polymerase chain reaction (PCR) identified diverse intestinal pathogens in Guatemalan children with diarrhea. Campylobacter, norovirus, sapovirus, and adenovirus were significant causes of illness in this low-income region.
Area of Science:
- Infectious Diseases
- Microbiology
- Public Health
Background:
- Low- and middle-income countries (LMICs) face persistent challenges with poverty, poor sanitation, and undernutrition, impacting child health.
- Intestinal pathogens contribute significantly to diarrheal diseases in children, particularly in resource-limited settings.
- Limited research exists on multiplex polymerase chain reaction (PCR) platforms for pathogen identification in Latin America.
Purpose of the Study:
- To characterize viral, bacterial, and parasitic enteric pathogens in children with acute diarrhea in Guatemala using multiplex PCR.
- To analyze correlations between identified pathogens and clinical, demographic, and socioeconomic factors.
- To describe pathogen acquisition, persistence, and clearance patterns and calculate population attributable fractions (PAFs) for diarrheal disease.
Main Methods:
- Multiplex PCR (BioFire) was employed to detect pathogens in stool samples from children aged 6-35 months with acute, non-bloody diarrhea.
- Stool samples were collected on day 1 and day 31 from participants in rural and urban Guatemala.
- Statistical analyses were performed to correlate pathogens with variables and calculate PAFs.
Main Results:
- A high prevalence of multiple pathogens per sample was observed, particularly in the rural setting (4.8 pathogens/sample vs. 2.7 in urban).
- Escherichia coli pathotypes, Shigella, Campylobacter, and Giardia showed high persistence rates over the 30-day study period.
- Significant adjusted PAFs for diarrheal causation were found for Campylobacter (14.9%), norovirus (10.2%), sapovirus (7.6%), and adenovirus 40/41 (5.6%).
Conclusions:
- Multiplex PCR platforms are valuable tools for understanding the complex landscape of enteric pathogens in children in LMICs.
- Patterns of symptomatic and asymptomatic enteric infections in Latin American children resemble those in other LMIC regions.
- Findings provide critical data for clinical management of infectious diarrhea and inform public health strategies in Guatemala and similar regions.
Abstract:
Multiplex polymerase chain reaction (PCR) platforms have enhanced understanding of intestinal pathogens in low- and middle-income countries (LMICs). However, few such studies have been performed in Latin America, where poverty, poor sanitation, and undernutrition persist. Multiplex PCR (BioFire, Salt Lake City, UT) was used to identify viral, bacterial, and parasitic pathogens in stool collected on day 1 and 31 from children aged 6 to 35 months with acute, non-bloody diarrhea in two locations (rural and urban) in Guatemala. We analyzed correlation between pathogens and clinical, demographic, and socioeconomic variables; described patterns of pathogen acquisition, persistence, and clearance over the 30-day period; and calculated population attributable fractions (PAFs) for diarrheal causation for individual pathogens. We analyzed 316 subjects (144 urban; 172 rural) enrolled between March 2015 and January 2016. Rural subjects had significantly more malnutrition, animal exposure, and unimproved water/sanitation infrastructure. The majority of subjects had multiple pathogens/sample (4.8 rural and 2.7 urban). Few meaningful correlates were identified between individual pathogens and clinical, demographic, or environmental variables. Escherichia coli pathotypes, Shigella, Campylobacter, and Giardia had high rates of persistence between initial and 30-day follow-up. Statistically significant adjusted PAFs were identified for Campylobacter (14.9%, 95% CI: 3.2-23.1), norovirus (10.2%, 95% CI: 0.4-17.1), sapovirus (7.6%, 95% CI: 2.3-10.9), and adenovirus 40/41 (5.6%, 95% CI: 0.3-8.7). These observations further characterize the diversity and complexity of enteric pathogens in children in LMICs. Patterns of chronic symptomatic and asymptomatic infection among Latin American children are similar to those observed in other LMIC regions. Findings have direct implications for practitioners treating individuals with acute infectious diarrhea and should inform regional public health strategies.
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