Department

De partment of Science, CWC, Department of Molecular Biology/Department of Botany, UW

Description

Hirschsprung’s disease (HD) is a congenital malformation of the gastrointestinal tract characterized by the absence of the distal enteric nervous system. Hirschsprung’s - associated enterocolitis (HAEC) is one of the most serious complications of HD. It is hypothesized t hat the intestinal microbiome contributes to the development of HAEC in human subjects. Our experimental design is to enroll five HD patients after they were diagnosed with HAEC. Fecal samples will be collected prior to antibiotic treatment, after completi on of antibiotic treatment, three months after treatment and six months after treatment. The samples will be frozen and pulverized. Extraction of genomic DNA will be performed by bead - beating technique which will be followed by phenol - chloroform extraction . Pyrosequencing will be outsourced. Then we will taxonomically classify and compare community structure between samples. We predict our results to show that the structure and function of the microbiome of each patient before antibiotic treatment is differ ent from those after treatment. This finding will support the hypothesis that disruption of normal intestinal microbiome contributes to the development of HAEC. These data will help to form strategies to positively impact the morbidity and mortality of HA EC.

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Contribution of the Intestinal Microbiome to the Development of Hirschsprung’s - Associated Enterocolitis

Hirschsprung’s disease (HD) is a congenital malformation of the gastrointestinal tract characterized by the absence of the distal enteric nervous system. Hirschsprung’s - associated enterocolitis (HAEC) is one of the most serious complications of HD. It is hypothesized t hat the intestinal microbiome contributes to the development of HAEC in human subjects. Our experimental design is to enroll five HD patients after they were diagnosed with HAEC. Fecal samples will be collected prior to antibiotic treatment, after completi on of antibiotic treatment, three months after treatment and six months after treatment. The samples will be frozen and pulverized. Extraction of genomic DNA will be performed by bead - beating technique which will be followed by phenol - chloroform extraction . Pyrosequencing will be outsourced. Then we will taxonomically classify and compare community structure between samples. We predict our results to show that the structure and function of the microbiome of each patient before antibiotic treatment is differ ent from those after treatment. This finding will support the hypothesis that disruption of normal intestinal microbiome contributes to the development of HAEC. These data will help to form strategies to positively impact the morbidity and mortality of HA EC.