A core-genome multilocus sequence typing scheme for the detection of genetically related Streptococcus pyogenes clusters

The recently observed increase in invasive infections causes concern in Europe. However, conventional molecular typing methods lack discriminatory power to aid investigations of outbreaks caused by . Therefore, there is an urgent need for high-resolution molecular typing methods to assess genetic relatedness between isolates. In the current study, we aimed to develop a novel high-resolution core-genome multilocus sequence typing (cgMLST) scheme for and compared its discriminatory power to conventional molecular typing methods. The cgMLST scheme was designed with the commercial Ridom SeqSphere+ software package. To define a cluster threshold, the scheme was evaluated using publicly available data from nine defined outbreaks in the United Kingdom. The cgMLST scheme was then applied to 23 isolates from a suspected outbreak and 117 . surveillance isolates both from the Netherlands. MLST and -typing results were used for comparison to cgMLST results. The allelic differences between isolates from defined outbreaks ranged between 6 and 31 for isolates with the same type, resulting in a proposed cluster threshold of 5 allelic differences out of 1,095 target loci. Seven out of twenty-three (30%) isolates from the suspected outbreak had an allelic difference of 2, thereby identifying a potential cluster that could not be linked to other isolates. The proposed cgMLST scheme shows a higher discriminatory ability when compared to conventional typing methods. The rapid and simple analysis workflow allows for extended detection of clusters of potential outbreak isolates and surveillance and may facilitate the sharing of sequencing results between (inter)national laboratories.