Heterogeneity of penicillin-non-susceptible group B streptococci isolated from a single patient in Germany.

OBJECTIVES: Streptococcus agalactiae [group B streptococci (GBS)] have been considered uniformly susceptible to penicillin. However, increasing reports from Asia and North America are documenting penicillin-non-susceptible GBS (PRGBS) with mutations in pbp genes. Here we report, to the best of our knowledge, the first two PRGBS isolates recovered in Europe (AC-13238-1 and AC-13238-2), isolated from the same patient. METHODS: Two different colony morphologies of GBS were noted from a surgical abscess drainage sample. Both were serotyped and antimicrobial susceptibility testing was performed by different methodologies. High-throughput sequencing was done to compare the isolates at the genomic level, to identify their capsular type and ST, to evaluate mutations in the pbp genes and to compare the isolates with the genomes of other PRGBS isolates sharing the same serotype and ST. RESULTS: Isolates AC-13238-1 and AC-13238-2 presented MICs above the EUCAST and CLSI breakpoints for penicillin susceptibility. Both shared the capsular type Ia operon and ST23. Genomic analysis uncovered differences between the two isolates in seven genes, including altered pbp genes. Deduced amino acid sequences revealed critical substitutions in PBP2X in both isolates. Comparison with serotype Ia clonal complex 23 PRGBS from the USA reinforced the similarity between AC-13238-1 and AC-13238-2, and their divergence from the US strains. CONCLUSIONS: Our results support the in-host evolution of ß-lactam-resistant GBS, with two PRGBS variants being isolated from one patient.

Evaluation of the 'HG Group B Streptococcus' loop-mediated isothermal amplification assay for accurate identification of Streptococcus agalactiae.

This study evaluated the HiberGene Group B Streptococcus test, a CE-IVD-approved molecular assay for rapid detection of Streptococcus agalactiae [Group B Streptococcus (GBS)] in human clinical specimens. Performance of the assay in terms of specificity, sensitivity and genotype inclusivity was investigated using an extended specificity panel of 113 human and animal GBS isolates, and eight isolates from other streptococcal species, from the isolate collection of the German National Reference Center for Streptococci. Broth cultures were tested according to the manufacturer's protocol, including lysis, heat denaturation and isothermal amplification. All 104/104 (100 %) human GBS isolates of nine serotypes (Ia, Ib, II, III, IV, V, VI, VII, VIII and non-typeable) were correctly identified by the assay as GBS. Additionally, 7/9 (78 %) GBS isolates from elephants were also correctly identified. Six isolates of other streptococcal species/subspecies (Streptococcus anginosus, S. constellatus, S. castoreus, S. dysgalactiae and S. dysgalactiae equisimilis) were correctly reported as negative. Two S. pyogenes (Group A Streptococcus) isolates gave invalid results. The HG Group B Streptococcus assay identified human GBS isolates in culture with 100 % sensitivity.