Genetic diversity of pneumococcal surface protein A (PspA) in paediatric isolates of non-conjugate vaccine serotypes in Japan

[[[ Purpose: ]]] Among the pneumococcal proteins, pneumococcal surface protein A (PspA) is considered the most promising candidate for a serotype-independent vaccine. This study aimed to investigate the serotype, genetic diversity of PspA, lineage (genotype) and drug resistance traits of pneumococcal isolates from paediatric patients. [[[ Methodology: ]]] A total of 678 non-invasive pneumococcal isolates obtained from June to November 2016 were analysed. All isolates were characterized for PspA families, serotypes and macrolide resistance genes. Seventy-one representative isolates of non-vaccine serotypes (NVTs) were genetically analysed for the clade-defining region (CDR) of PspA, as well as multi-locus sequence typing (MLST). [[[ Results: ]]] The detection rate of NVTs was 87.9 % (n=596), including dominant NVTs 15A (14.5 %, n=98), 35B (11.8 %, n=80), 15C (9.3 %, n=63) and 23A (9.0 %, n=61). Most isolates (96.6 %) possessed macrolide resistance genes erm(B) and/or mef(A/E). PspA families 1, 2 and 3 were detected in 42.3, 56.6 and 0.6 % of isolates, respectively. Nucleotide sequences of CDR showed high identity (90-100 %) within the same PspA clade, although the CDR identity among different PspA families ranged from 53 to 69 %. All isolates of NVTs 23A, 10A, 34, 24, 22F/22A, 33F, 23B and 38 were from PspA family 1, while NVTs 35B, 15C, 15B and 11A/11D isolates were from family 2. In contrast, genetically distinct PspAs were found in NVTs 6C and 15A. PspA family 3/clade 6 was detected in only NVT serotype 37 isolates assigned to ST447 and ST7970, showing the mucoid phenotype. [[[ Conclusion: ]]] The present study revealed the predominance of PspA families 1 and 2 in NVTs, and the presence of family 3 in serotype 37.