Streptococcus suis is an important bacterial pathogen in pigs that may also cause zoonotic disease in humans. The aim of the study was to evaluate MALDI-TOF MS identification of S. suis case isolates from diseased pigs and tonsil isolates from healthy pigs and wild boar, using sequence analysis methods. Isolates (n=348) which had been classified as S. suis by MALDI-TOF MS were whole-genome sequenced and investigated using analysis of i) the 16S rRNA gene, ii) the recN gene, and iii) whole-genome average nucleotide identity (ANI). Analysis of the 16S rRNA gene indicated that 82.8% (288 out of 348) of the isolates were S. suis, while recN-gene analysis indicated that 75.6% (263 out of 348) were S. suis. ANI analysis classified 44.3% (154 out of 348) as S. suis. In total, 44% (153 out of 348) of the investigated isolates were classified as S. suis by all of the species identification methods employed. The mean MALDI-TOF MS score was significantly higher for the S. suis case isolates compared to the tonsil isolates, however, the difference is of limited practical use. The results show that species confirmation beyond MALDI-TOF MS is needed for S. suis isolates. For this purpose, ANI analysis with a slightly lowered cutoff of 94% may be used instead of, or in addition to, recN-gene analysis. Supplementation of the MALDI-TOF MS reference library with mass spectra from S. orisratti, S. parasuis, S. ruminantium, and additional S. suis serotypes, should be considered in order to produce more accurate classifications.
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