Novel staphylococci nucH taxonomical marker used in identification of human-associated Staphylococcus succinus subsp. casei.

The aim of our study was to assess the sequencing of unique nucH gene fragment based on performed bioinformatics analysis as a novel diagnostic method for the identification of difficult to identify staphylococcal human pathogenic strains. Initially, PCR-RFLP-rrn analysis specific to the spacers between 16SrDNA and 23SrDNA followed by HhaI restriction analysis was performed. Further, sequencing of nucH and 16S rDNA genes fragments was carried out. Blast analysis from the NCBI showed 99% similarity of nucH gene fragment with reference genomic DNA for S. succinus with the accession no. CP018199. This result was also confirmed by MALDI-TOF analysis. Sequencing analysis of 16S rDNA gene fragment allowed for 100% identification of two strains isolated from human samples as Staphylococus succinus subsp. casei. Sequencing of identified unique nucH gene fragment seems to be a promising diagnostic assay for the identification of Staphylococcus species. Based on our results, we can assume that probably other Staphylococcus species originated from different clinical samples could be identified using nucH gene sequencing method we developed. However, an extension of the genetic databases with a substantially bigger number of reference staphylococcal species for nucH gene is needed to make this method better than widely used standard 16S rDNA sequencing assay. To the best of our knowledge, it is the second published isolation of S. succinus subsp. casei from human clinical specimens. Moreover, possibility of decreasing the number of dimensions from multi-PCR-bands results using ribotyping analysis is also described.