lytA-based identification methods can misidentify Streptococcus pneumoniae

During surveillance studies we detected, among over 1500 presumptive pneumococci, 11 isolates displaying conflicting or novel results when characterized by widely accepted phenotypic (optochin susceptibility and bile solubility) and genotypic (lytA-BsaAI-RFLP and MLST) identification methods. We aimed to determine the genetic basis for the unexpected results given by lytA-BsaAI-RFLP and investigate the accuracy of the WHO recommended lytA real-time PCR assay to classify these 11 isolates. Three novel lytA-BsaAI-RFLP signatures were found (one in pneumococcus and two in S. mitis). In addition, one pneumococcus displayed the atypical lytA-BsaAI-RFLP signature characteristic of non-pneumococci and two S. pseudopneumoniae displayed the typical lytA-BsaAI-RFLP pattern characteristic of pneumococci. lytA real-time PCR misidentified these three isolates. In conclusion, identification of pneumococci by lytA real-time PCR, and other lytA-based methodologies, may lead to false results. This is of particular relevance in the increasingly frequent colonization studies relying solely on culture-independent methods. [Display omitted] •Identification of pneumococci by lytA-based methodologies may lead to false results.•A pneumococcal carriage strain harbouring a lytA homologue is described.•Two invasive S. pseudopneumoniae harbouring the pneumococcal lytA are described.•These strains are misidentified by the WHO recommended lytA-real time PCR assay.•Three novel lytA-BsaAI-RFLP signatures are described.