Dataset Information

Phenotypic and molecular analysis of nontypeable Group B streptococci: identification of cps2a and hybrid cps2a/cps5 Group B streptococcal capsule gene clusters.

ABSTRACT: The Group B streptococcus (GBS) can express a capsular polysaccharide (CPS). There are ten recognized CPSs (Ia, Ib, and II-IX). A GBS isolate is considered nontypeable (NT) when CPS cannot be identified as one of ten types. Two groups of GBS NT isolates were studied, isolates without surface sialic acid (sia(-)) and isolates with surface sialic acid (sia(+)). The first objective was to characterize NT sia(-) isolates that failed CPS identification by an immunodiffusion antisera typing assay and a RT-PCR capsule typing assay. NT sia(-) isolates were characterized by assaying phenotypic changes and identifying covR/S mutations that may potentially have a role in the altered phenotypes. The second objective was to characterize NT sia(+) isolates that failed to identify as one of the ten CPS types by an immundiffusion antisera-based typing assay and a RT-PCR capsule typing assay yet expressed capsule. Fifteen NT sia(-) isolates displayed increased ? hemolysis/orange pigmentation, decreased CAMP activity, inability to form biofilm, and susceptibility to phagocytosis by human blood. DNA sequence analysis of the covR/S genes in the sia(-) isolates found mutations in 14 of 15 isolates assayed. These mutations in the covR/S genes may potentially contribute to lack of expression of phenotypic traits assayed in vitro. For the three NT sia(+) isolates, whole-genome sequence analyses identified two isolates with cps gene clusters identical to the recently described and uncommon CPSIIa type. The third isolate possessed a hybrid cluster containing cps genes for both CPSIIa and CPSV suggesting recombination between these two gene clusters.

SUBMITTER: Alhhazmi A

PROVIDER: S-EPMC6081472 | biostudies-literature | 2018 Aug

REPOSITORIES: biostudies-literature

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Phenotypic and molecular analysis of nontypeable Group B streptococci: identification of cps2a and hybrid cps2a/cps5 Group B streptococcal capsule gene clusters.

Alhhazmi Areej A Tyrrell Gregory J GJ

Emerging microbes & infections 20180808 1

The Group B streptococcus (GBS) can express a capsular polysaccharide (CPS). There are ten recognized CPSs (Ia, Ib, and II-IX). A GBS isolate is considered nontypeable (NT) when CPS cannot be identified as one of ten types. Two groups of GBS NT isolates were studied, isolates without surface sialic acid (sia(-)) and isolates with surface sialic acid (sia(+)). The first objective was to characterize NT sia(-) isolates that failed CPS identification by an immunodiffusion antisera typing assay and ...[more]

PMID: 30087323

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Project description:Streptococcus agalactiae or group B streptococcus (GBS) is a leading cause of serious neonatal infections. GBS is an opportunistic commensal constituting a part of the intestinal and vaginal physiologic flora and maternal colonization is the principal route of GBS transmission. GBS is a pathobiont that converts from the asymptomatic mucosal carriage state to a major bacterial pathogen causing severe invasive infections. At present, as many as 10 serotypes (Ia, Ib, and II-IX) are recognized. The aim of the current review is to shed new light on the latest epidemiological data and clonal distribution of GBS in addition to discussing the most important colonization determinants at a molecular level. The distribution and predominance of certain serotypes is susceptible to variations and can change over time. With the availability of multilocus sequence typing scheme (MLST) data, it became clear that GBS strains of certain clonal complexes possess a higher potential to cause invasive disease, while other harbor mainly colonizing strains. Colonization and persistence in different host niches is dependent on the adherence capacity of GBS to host cells and tissues. Bacterial biofilms represent well-known virulence factors with a vital role in persistence and chronic infections. In addition, GBS colonization, persistence, translocation, and invasion of host barriers are largely dependent on their adherence abilities to host cells and extracellular matrix proteins (ECM). Major adhesins mediating GBS interaction with host cells include the fibrinogen-binding proteins (Fbs), the laminin-binding protein (Lmb), the group B streptococcal C5a peptidase (ScpB), the streptococcal fibronectin binding protein A (SfbA), the GBS immunogenic bacterial adhesin (BibA), and the hypervirulent adhesin (HvgA). These adhesins facilitate persistent and intimate contacts between the bacterial cell and the host, while global virulence regulators play a major role in the transition to invasive infections. This review combines for first time epidemiological data with data on adherence and colonization for GBS. Investigating the epidemiology along with understanding the determinants of mucosal colonization and the development of invasive disease at a molecular level is therefore important for the development of strategies to prevent invasive GBS disease worldwide.

Project description:The major virulence determinant of group A streptococci is the ability to resist opsonization and phagocytic ingestion. The present studies were performed to compare the mechanisms of resistance to opsonization of type 18 and type 24 streptococci and to determine the relative roles of M protein-fibrinogen interaction and the hyaluronate capsule in preventing phagocytic ingestion and killing. By use of parent strains and acapsular transposon mutants in the presence and absence of fibrinogen, we show that type 18 and type 24 streptococci rely on somewhat different mechanisms for resistance to opsonization. Type 24 streptococci bound fibrinogen avidly to their surfaces, and encapsulated organisms were completely resistant to opsonization only in the presence of fibrinogen. In contrast, type 18 streptococci bound 10-fold less fibrinogen than type 24 streptococci and were fully resistant to phagocytosis only when they expressed capsule. The general structural characteristics of the amino-terminal halves of type 18 and type 24 M proteins differed in that type 18 M protein contained only one complete B repeat, whereas type 24 M protein contained five complete B repeats, a structural difference which could potentially be related to the differences in fibrinogen binding between the two serotypes. Immunofluorescence assays of complement deposition were used in combination with 125I-C3 binding assays to show that encapsulated type 24 streptococci were fully resistant to opsonization by C3 only in the presence of plasma. Encapsulated and unencapsulated type 18 streptococci were equally opsonized by C3 in either plasma or serum, yet only encapsulated organisms resisted phagocytic killing in blood. The results of this study indicate that opsonization by C3 does not necessarily lead to phagocytic ingestion and that the hyaluronate capsule and M proteins are variably important in resistance to different group A streptococci to opsonization and phagocytic killing.

Dataset Information

Phenotypic and molecular analysis of nontypeable Group B streptococci: identification of cps2a and hybrid cps2a/cps5 Group B streptococcal capsule gene clusters.

Publications

Phenotypic and molecular analysis of nontypeable Group B streptococci: identification of cps2a and hybrid cps2a/cps5 Group B streptococcal capsule gene clusters.

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