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The human oral microbiome has a major role in oral diseases including dental caries. Our studies on progression of caries infection through dentin and more recently, the invasion of vital dental pulp, detected  Lactobacillus rhamnosus  in the initial stages of infection of vital pulp tissue. In this study employing current high-throughput next generation sequencing technology we sought to obtain insight into genomic traits of tissue invasive  L. rhamnosus , to recognise biomarkers that could provide an understanding of pathogenic potential of lactobacilli, generally regarded as safe. Roche GS FLX+ technology was used to generate whole genome sequences of two clinical isolates of  L. rhamnosus  infecting vital pulp. Detailed genome-wide comparison of the genetic profiles of tissue invasive  L. rhamnosus  with probiotic  L. rhamnosus  was performed to test the hypothesis that specific strains of  L. rhamnosus  possessing a unique gene complement are selected for the capacity to invade vital pulp tissue. Analysis identified 264 and 258 genes respectively, from dental pulp-invasive  L. rhamnosus  strains LRHMDP2 and LRHMDP3 isolated from two different subjects that were not present in the reference probiotic  L. rhamnosus  strain ATCC 53103 (GG). Distinct genome signatures identified included the presence of a modified exopolysaccharide cluster, a characteristic confirmed in a further six clinical isolates. Additional features of LRHMDP2 and LRHMDP3 were altered transcriptional regulators from RpoN, NtrC, MutR, ArsR and zinc-binding Cro/CI families, as well as changes in the two-component sensor kinase response regulator and ABC transporters for ferric iron. Both clinical isolates of  L. rhamnosus  contained a single  Spa FED cluster, as in  L. rhamnosus  Lc705, instead of the two  Spa  clusters ( Spa CBA and  Spa FED) identified in  L. rhamnosus  ATCC 53103 (GG). Genomic distance analysis and SNP divergence confirmed a close relationship of the clinical isolates but segregation from the reference probiotic  L. rhamnosus  strain ATCC 53103 (GG).

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Comparative Genome Analysis of Lactobacillus rhamnosus Clinical Isolates from Initial Stages of Dental Pulp Infection: Identification of a New Exopolysaccharide Cluster