ORALMET: The Oral Microbiota of Irish Children

The oral cavity is a complex niche. The microorganisms found in the oral cavity are referred to as the oral microbiota and the collection of genes that these bacteria carry is the oral microbiome. This oral microbiome is intimately related to health and disease. With the development of metagenomic techniques, it is now possible to study complete bacterial communities, which was not previously possible using culture-dependent methods.

The aim of the ORALMET project is to identify oral microbiota changes that occur from birth to pre-school age in a longitudinally-studied cohort, and to compare this data with baseline microbiota analyses of caries-affected and caries-free children. The first objective of this study is to characterise the developing oral microbiota from birth to 36 months using microbiota profiling. The second overlapping objective is to investigate the oral microbiota in a cohort of children from 12-71 months, to generate insight into the microbiological risk factors for disease progression and which may lead to possible interventions.

Method:

Mothers were recruited antenatally at Cork University Maternity Hospital. Skin swabs from c-Section mothers and Vaginal swabs from Natural delivered mothers were taken, and saliva swabs from all mothers and their babies at different time-points. We compared and validated DNA extraction methods for saliva samples from mothers and babies, and for vaginal/skin swabs from mothers. DNA extraction methods were validated, PCR of the 16s rRNA V4/5 region was completed. Samples were then purified using Agencourt AMPure XP, Beckman Coulter. The samples were then Quantified using Quant-iT™ PicoGreen® dsDNA Assay Kit, Life Technologies. After Pooling, samples were sent for MiSeq Sequencing.

Result:

Results of this MiSeq sequencing to be presented.

Conclusion:

Following comparison of DNA extraction methods, genomic DNA is now being extracted with all samples using the MO Bio kit, and following the above sequencing techniques.