Discovery of a <i>Streptococcus pneumoniae</i> serotype 33F capsular polysaccharide locus that lacks <i>wcjE</i> and contains a <i>wcyO</i> pseudogene

<div><p>As part of large on-going vaccine impact studies in Fiji and Mongolia, we identified 25/2750 (0.9%) of nasopharyngeal swabs by microarray that were positive for <i>Streptococcus pneumoniae</i> contained pneumococci with a divergent 33F capsular polysaccharide locus (designated ‘33F-1’). We investigated the 33F-1 capsular polysaccharide locus to better understand the genetic variation and its potential impact on serotyping results. Whole genome sequencing was conducted on ten 33F-1 pneumococcal isolates. Initially, sequence reads were used for molecular serotyping by PneumoCaT. Phenotypic typing of 33F-1 isolates was then performed using the Quellung reaction and latex agglutination. Genome assemblies were used in phylogenetic analyses of each gene in the capsular locus to investigate genetic divergence. All ten pneumococcal isolates with the 33F-1 <i>cps</i> locus typed as 33F by Quellung and latex agglutination. Unlike the reference 33F capsule locus sequence, DNA microarray and PneumoCaT analyses found that 33F-1 pneumococci lack the <i>wcjE</i> gene, and instead contain <i>wcyO</i> with a frameshift mutation. Phylogenetic analyses found the <i>wzg</i>, <i>wzh</i>, <i>wzd</i>, <i>wze</i>, <i>wchA</i>, <i>wciG</i> and <i>glf</i> genes in the 33F-1 <i>cps</i> locus had higher DNA sequence similarity to homologues from other serotypes than to the 33F reference sequence. We have discovered a novel genetic variant of serotype 33F, which lacks <i>wcjE</i> and contains a <i>wcyO</i> pseudogene. This finding adds to the understanding of molecular epidemiology of pneumococcal serotype diversity, which is poorly understood in low and middle-income countries.</p></div>