Development of oral microflora of infants in relation to oral and general health: longitudinal analysis of babies under an oral Health Program for early childhood and their relative s microflora by pyrosequencing

Abstract

Defining the oral microbioma of infants and how it changes over time would be of great importance for the prevention, diagnosis and treatment of oral diseases at an early stage. The aim of this study is to evaluate the microbial colonization of children attending an oral health program for infants at 6, 12, 18 and 24 months and the microbial diversity associated with diet, oral hygiene, presence of teeth, tooth decay, periodontal conditions compared to oral conditions and microbial diversity of the parents / guardians. Eighty healthy babies will be monitored at 6, 12, 18 and 24 months. Food intake and oral hygiene will be evaluated by a questionnaire. Saliva and biofilm from babys and parents / guardians will be collected for analysis of microbial diversity by pyrosequencing. Assessment of gingival condition and caries prevalence of children and caregivers and the chronology of tooth eruption of the baby will be also held. Five healthy babies will be selected for the molecular analysis. After the extraction and quantitation of DNA, a library of PCR products of the hypervariable region of the genes V5-V6 will be generated for each sample. The amplicons libraries will be sequenced unidirectionally in the reverse direction by Genome Sequencing System FLX (GS FLX). The algorithm GAST will be used to calculate the percentage difference between each sequence and its closest match to a database of sequences V5-V6 Eubacteria and Archaea. The taxonomic groups will be designated for each complete reference sequence, using a variety of sources. The Shannon Diversity Index and Index of Principal Component Analysis will be obtained using the program PAST. (AU)