Identification of the anginosus group within the genusStreptococcususing polymerase chain reaction

Abstract

The aim of this study was to establish an identification method for the anginosus group within the genus Streptococcus by polymerase chain reaction (PCR). Using a primer pair based on the group-specific sequences of penicillin-binding protein 2B (pbp2b) gene, a 275-bp fragment was amplified from each species in the group but no size-matched products were obtained in other streptococci. Further identification in the species or subspecies level was possible by a multiplex PCR with primers for the 16S ribosomal RNA gene of Streptococcus anginosus, the hyaluronate lyase genes both of Streptococcus intermedius and Streptococcus constellatus subsp. constellatus, and the intermedilysin (ily) gene of S. intermedius. In the case ofStreptococcus constellatus subsp. pharyngis, the amplified fragment from the S. intermedius-type hyaluronate lyase gene was obtained, while that from the ily gene was not. These results also indicate that two different hyaluronate lyase genes are distributed among the anginosus group.

Streptococcus anginosus (milleri) Group Strains Isolated in Poland (1996–2012) and their Antibiotic Resistance Patterns

“…So, even with advanced techniques used for strain identification, we experienced multiple problems with proper species identification. In laboratories that routinely perform identification using PCR methods, detection and differentiation of SAG can be performed using molecular methods (Takao et al, 2004). From the clinical point of view, identification to anginosus group is enough to consider an isolate as an etiological factor of infection, however, identification as S. gordonii, which is a natural oral flora, may lead to misdiagnosis and not considering the strain as source of the infection.…”

“…Unfortunately, even with the use of 16S rDNA sequencing, some of the strains cannot be clearly classified to previously described streptococcal species (Olson et al, 2013;Thompson et al, 2013). There are some methods available to discriminate streptococcal strains to the species and subspecies level (Poyart et al, 1998;Picard et al, 2004;Glazunova et al, 2010;Zbinden et al, 2011;Obszanska et al, 2015a;Takao et al, 2004). However, sequencing methods are not often used in diagnostic laboratory as the routine spe cies identification and not all microbiology diagnostic laboratories have PCR set up.…”

Novel Twin Streptolysin S-Like Peptides Encoded in the sag Operon Homologue of Beta-Hemolytic Streptococcus anginosus

“…The type strain, NCTC10713, was used for molecular analyses of the SLS homologue of S. anginosus, and a further 125 strains of S. anginosus from human clinical sources were tested for epidemiological purposes. Initial confirmation of these strains as belonging to S. anginosus was by the PCR-based method of Takao et al (12). The type strain of S. pyogenes GTC262 and a streptolysin O gene (slo)-deleted mutant of S. pyogenes strain NIH35 were also used for the comparison of hemolytic characteristics of SLS and SLS homologues.…”

A streptolysin S homologue is essential for β-haemolytic Streptococcus constellatus subsp. constellatus cytotoxicity

“…The presence of sagA SCC was confirmed by PCR amplification using primers designed against the full-length sagA SCC from strain W277. The PCR amplification was conducted using Ex Taq DNA polymerase (TaKaRa) with the SCC_sagA-Fw1 and SCC_sagA-Bw1 primer set (Table 1) and the bacterial cell suspension as templates, which were prepared as described previously (Takao et al, 2004). The reaction mixture was incubated for 5 min at 94 uC and then subjected to 40 cycles using the following conditions: 98 uC for 10 s, 55 uC for 30 s and 68 uC for 20 s. Finally, the mixture was heated once at 72 uC for 5 min.…”