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The LuxS protein, encoded by <i>luxS</i>, is required for the production of autoinducer 2 (AI-2) in <i>Streptococcus pneumoniae</i>. The AI-2 molecule serves as a quorum sensing signal, and thus regulates cellular processes such as carbohydrate utilisation and biofilm formation, as well as impacting virulence. The role of <i>luxS</i> in <i>S. pneumoniae</i> biology and lifestyle has been predominantly assessed in the laboratory strain D39. However, as biofilm formation, which is regulated by <i>luxS</i>, is critical for the ability of <i>S. pneumoniae</i> to cause otitis media, we investigated the role of <i>luxS</i> in a middle ear isolate, strain 947. Our results identified <i>luxS</i> to have a role in prevention of <i>S. pneumoniae</i> transition from colonisation of the nasopharynx to the ear, and in facilitating adherence to host epithelial cells.