Pneumococcal Research, Murdoch Childrens Research Institute, Royal Children's Hospital, Parkville, Victoria, Australia.
Pneumococcal Research, Murdoch Childrens Research Institute, Royal Children's Hospital, Parkville, Victoria, Australia.
Clin Microbiol Infect. 2018 Apr;24(4):428.e1-428.e7. doi: 10.1016/j.cmi.2017.06.031. Epub 2017 Jul 20.
As part of annual cross-sectional Streptococcus pneumoniae carriage surveys in Fiji (2012-2015), we detected pneumococci in over 100 nasopharyngeal swabs that serotyped as '11F-like' by microarray. We examined the genetic basis of this divergence in the 11F-like capsular polysaccharide (cps) locus compared to the reference 11F cps sequence. The impact of this diversity on capsule phenotype, and serotype results using genetic and serologic methods were determined.
Genomic DNA from representative 11F-like S. pneumoniae isolates obtained from the nasopharynx of Fijian children was extracted and subject to whole genome sequencing. Genetic and phylogenetic analyses were used to identify genetic changes in the cps locus. Capsular phenotypes were evaluated using the Quellung reaction and latex agglutination.
Compared to published 11F sequences, the wcwC and wcrL genes of the 11F-like cps locus are phylogenetically divergent, and the gct gene contains a single nucleotide insertion within a homopolymeric region. These changes within the DNA sequence of the 11F-like cps locus have modified the antigenic properties of the capsule, such that 11F-like isolates serotype as 11A by Quellung reaction and latex agglutination.
This study demonstrates the ability of molecular serotyping by microarray to identify genetic variants of S. pneumoniae and highlights the potential for discrepant results between phenotypic and genotypic serotyping methods. We propose that 11F-like isolates are not a new serotype but rather are a novel genetic variant of serotype 11A. These findings have implications for invasive pneumococcal disease surveillance as well as studies investigating vaccine impact.
作为斐济年度横断面肺炎链球菌携带调查的一部分(2012-2015 年),我们通过微阵列检测到超过 100 份鼻咽拭子中的肺炎球菌,这些肺炎球菌经微阵列检测为“11F 样”。我们比较了参考 11F 荚膜多糖(cps)序列与“11F 样”荚膜多糖(cps)基因座的遗传基础,研究了这种差异的原因。通过遗传和血清学方法确定了这种多样性对荚膜表型和血清型结果的影响。
从斐济儿童鼻咽部获得的代表 11F 样 S. pneumoniae 分离株的基因组 DNA 提取后进行全基因组测序。利用遗传和系统发育分析确定 cps 基因座的遗传变化。用凝固酶反应和乳胶凝集法评估荚膜表型。
与已发表的 11F 序列相比,“11F 样”cps 基因座的 wcwC 和 wcrL 基因在系统发育上是不同的,gct 基因在一个同源多聚区域内有一个单核苷酸插入。cps 基因座 DNA 序列中的这些变化改变了荚膜的抗原特性,使得 11F 样分离株通过凝固酶反应和乳胶凝集反应血清型为 11A。
本研究表明,微阵列分子血清分型能够识别肺炎链球菌的遗传变异,并强调了表型和基因型血清分型方法之间可能存在不一致的结果。我们提出,11F 样分离株不是一个新的血清型,而是血清型 11A 的一个新的遗传变异。这些发现对侵袭性肺炎球菌病监测以及研究疫苗影响具有重要意义。