Clustered Regularly Interspaced Short Palindromic Repeat Analysis of Clonal Complex 17 Serotype III Group B Streptococcus Strains Causing Neonatal Invasive Diseases

Abstract

Group B Streptococcus (GBS) is an important pathogen of neonatal infections, and the clonal complex (CC)-17/serotype III GBS strain has emerged as the dominant strain. The clinical manifestations of CC17/III GBS sepsis may vary greatly but have not been well-investigated. A total of 103 CC17/III GBS isolates that caused neonatal invasive diseases were studied using a new approach based on clustered regularly interspaced short palindromic repeats (CRISPR) loci and restriction fragment length polymorphism (RFLP) analyses. All spacers of CRISPR loci were sequenced and analyzed with the clinical presentations. After CRISPR-RFLP analyses, a total of 11 different patterns were observed among the 103 CRISPR-positive GBS isolates. GBS isolates with the same RFLP patterns were found to have highly comparable spacer contents. Comparative sequence analysis of the CRISPR1 spacer content revealed that it is highly diverse and consistent with the dynamics of this system. A total of 29 of 43 (67.4%) spacers displayed homology to reported phage and plasmid DNA sequences. In addition, all CC17/III GBS isolates could be categorized into three subgroups based on the CRISPR-RFLP patterns and eBURST analysis. The CC17/III GBS isolates with a specific CRISPR-RFLP pattern were more significantly associated with occurrences of severe sepsis (57.1% vs. 29.3%, p = 0.012) and meningitis (50.0% vs. 20.8%, p = 0.009) than GBS isolates with RFLP lengths between 1000 and 1300 bp. Whole-genome sequencing was also performed to verify the differences between CC17/III GBS isolates with different CRISPR-RFLP patterns. We concluded that the CRISPR-RFLP analysis is potentially applicable to categorizing CC17/III GBS isolates, and a specific CRISPR-RFLP pattern could be used as a new biomarker to predict meningitis and illness severity after further verification.

Current research update on group B streptococcal infection related to obstetrics and gynecology

“…ALP family proteins are commonly expressed virulence factors in GBS that are also associated with immune evasion (Paoletti and Kasper, 2019;Lin L. et al, 2021). This family of proteins includes ALP-C, ALP-1, ALP-2, ALP-3, ALP-4, and Rib, encoded by the genes bca, alp1, alp2, alp3, alp4, and rib, respectively, and their amino acid sequences exhibit homology (Furfaro et al, 2018).…”

“…Currently, there is limited understanding of the pathogenesis of late-onset GBS infection ( Delara et al, 2023 ), which may be related to vertical transmission, nosocomial infection, or community-acquired infection. Serotype III GBS is highly associated with meningitis ( Hsu et al, 2021 ). Recent studies have shown that preterm birth is a major risk factor for late-onset GBS infection ( Gonçalves et al, 2022 ; Choi et al, 2023 ; Paul et al, 2023 ).…”

CRISPR/Cas9 Technology and Its Utility for Crop Improvement

“…When dCas binds to the promoter or transcription start site of the target gene, it can prevent the initiation of transcription. When dCas binds to the target gene open reading frame, it prevents the binding of RNA polymerases and transcription factors, thereby inhibiting transcription [ 46 ].…”

“…Ma et al successfully reduced the amylose content of rice grains from 14.6% to 2.6% by knocking out the WAXY ( Wx ) gene, thereby obtaining waxy rice mutant plants [ 25 ]. Similarly, the maize Wx1 gene encodes a starch synthase affecting the grain composition [ 46 ]. Knocking out Wx1 via the CRISPR/Cas9 system can increase the amylopectin content of maize grains to almost 100% [ 47 ], without inducing other phenotypic changes.…”

An Application of Real-Time PCR and CDC Protocol May Significantly Reduce the Incidence of Streptococcus agalactiae Infections among Neonates

“…Streptococcus agalactiae identification is a challenge for modern microbiological diagnostics, especially taking into account the phenotypic and molecular diversity of the strains belonging to this species [7][8][9][10]. Moreover, in recent years, an increase in GBS resistance to selected drugs has been observed [11].…”