Identification of Enteropathogens by Multiplex PCR among Rural and Urban Guatemalan Children with Acute Diarrhea

James T. Gaensbauer Center for Global Health and Department of Epidemiology, Colorado School of Public Health, Aurora, Colorado;
Department of Pediatrics, Denver Health Hospital Authority, Denver, Colorado;
Department of Pediatrics, University of Colorado School of Medicine, Aurora, Colorado;

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Molly Lamb Center for Global Health and Department of Epidemiology, Colorado School of Public Health, Aurora, Colorado;

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Diva M. Calvimontes Hospital Roosevelt, Guatemala City, Guatemala;

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Edwin J. Asturias Center for Global Health and Department of Epidemiology, Colorado School of Public Health, Aurora, Colorado;
Department of Pediatrics, University of Colorado School of Medicine, Aurora, Colorado;

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Satoshi Kamidani Department of Pediatrics, University of Colorado School of Medicine, Aurora, Colorado;

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Ingrid L. Contreras-Roldan Centro de Estudios en Salud, Universidad de Valle de Guatemala, Guatemala, Guatemala;

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Samuel R. Dominguez Department of Pediatrics, University of Colorado School of Medicine, Aurora, Colorado;
Department of Pathology and Laboratory Medicine, Children’s Hospital Colorado, Aurora, Colorado

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Christine C. Robinson Department of Pathology and Laboratory Medicine, Children’s Hospital Colorado, Aurora, Colorado

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Alma Zacarias Center for Global Health and Department of Epidemiology, Colorado School of Public Health, Aurora, Colorado;

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Stephen Berman Center for Global Health and Department of Epidemiology, Colorado School of Public Health, Aurora, Colorado;
Department of Pediatrics, University of Colorado School of Medicine, Aurora, Colorado;

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Mario A. Melgar Hospital Roosevelt, Guatemala City, Guatemala;

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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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Author Notes

Address correspondence to James T. Gaensbauer, Denver Health Hospital Authority, Pavilion C, MC 0590, 777 Bannock St., Denver, CO 80204. E-mail: james.gaensbauer@dhha.org

Disclosure: E. J. A. reports grants from Pantheryx, G. S. K., and Pfizer outside the submitted work. S. B., I. L. C. R., J. T. G., M. L., and C. C. R., report grants from Pan Theryx, during the conduct of the study. M. A. M. reports personal fees from Fundacion para la salud integral de los guatemaltecos during the conduct of the study. S. R. D. reports grants from Biofire during the conduct of the study.

Financial support: Funding for the clinical trial from which data were obtained was provided by Pan Theryx, Inc., Boulder, CO. No funding was received for this analysis.

Authors’ addresses: James T. Gaensbauera, Center for Global Health and Department of Epidemiology, Colorado School of Public Health, Aurora, CO, Department of Pediatrics, Denver Health Hospital Authority, Denver, CO, and Department of Pediatrics, University of Colorado School of Medicine, Aurora, CO, E-mail: james.gaensbauer@dhha.org. Molly Lamb and Alma Zacarias, Center for Global Health and Department of Epidemiology, Colorado School of Public Health, Aurora, CO, E-mails: molly.lamb@cuanschutz.edu and almaloarca@yahoo.es. Diva M. Calvimontes and Mario A. Melgar, Hospital Roosevelt, Guatemala City, Guatemala, E-mails: mirellacalvimontes@yahoo.com and mariomelgart@gmail.com. Edwin J. Asturias and Stephen Berman, Center for Global Health and Department of Epidemiology, Colorado School of Public Health, Aurora, CO, and Department of Pediatrics, University of Colorado School of Medicine, Aurora, CO, E-mails: edwin.asturias@childrenscolorado.org and stephen.berman@childrenscolorado.org. Satoshi Kamidani, Department of Pediatrics, University of Colorado School of Medicine, Aurora, CO, E-mail: satoshi.kamidani@emory.edu. Ingrid L. Contreras-Roldan, Centro de Estudios en Salud, Universidad de Valle de Guatemala, Guatemala, Guatemala, E-mail: icontreras@ces.uvg.edu.gt. Samuel R. Dominguez, Department of Pediatrics, University of Colorado School of Medicine, Aurora, Colorado, and Department of Pathology and Laboratory Medicine, Children’s Hospital Colorado, Aurora, CO, E-mail: samuel.dominguez@childrenscolorado.org. Christine C. Robinson, Department of Pathology and Laboratory Medicine, Children’s Hospital Colorado, Aurora, CO, E-mail: christine.robinson@childrenscolorado.org.

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