title: Superantigen gene complement of Streptococcus pyogenes--relationship with other typing methods and short-term stability.
abstract: The profiling of the superantigen (SAg) encoding genes has been frequently used as a complementary typing method for group A streptococci (GAS), but a confusing gene nomenclature and a large diversity of primers used in screening has led to some conflicting results. The aim of this work was to develop a polymerase chain reaction (PCR) method capable of efficiently amplifying all the known allelic variants of these genes, and to evaluate the congruence of this methodology with other commonly used molecular typing methods. The presence of the 11 known SAg genes and two other exotoxin-encoding genes (speB and speF) was tested in a collection of 480 clinical GAS isolates, using two multiplex PCR reactions. The SAg gene profile was compared with other typing methods. Four naturally occurring deletions involving the genes speB, speF, and rgg were characterized, two of which were found among invasive isolates. The absence of the chromosomally encoded genes speG and smeZ was supported by Southern blot hybridization and associated with specific GAS lineages, while the presence of phage-encoded genes was more variable. Positive associations between SAg genes or between SAg profiles and emm types or pulsed-field gel electrophoresis (PFGE) clusters were observed. The results suggest that the SAg profile diversifies faster than other properties commonly used for molecular typing, such as emm type and multilocus sequence typing (MLST) sequence types (STs), and can be a useful complement in GAS molecular epidemiology. Still, the short-term stability of the SAg gene profile among prevalent genetic lineages may largely explain the observed associations between SAg genes.
authors.affiliation: Instituto de Microbiologia, Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Av Prof Egas Moniz, 1649-028 Lisboa, Portugal.
authors.forename: A
authors.initials: A
authors.lastname: Friães
authors.forename: F R
authors.initials: FR
authors.lastname: Pinto
authors.forename: C
authors.initials: C
authors.lastname: Silva-Costa
authors.forename: M
authors.initials: M
authors.lastname: Ramirez
authors.forename: J
authors.initials: J
authors.lastname: Melo-Cristino
mesh_terms: D000942:Antigens, Bacterial / Q000235:genetics; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D015139:Blotting, Southern; D002352:Carrier Proteins / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field; D017353:Gene Deletion; D005798:Genes, Bacterial; D042822:Genomic Instability; D006801:Humans; D058889:Molecular Typing / Q000379:methods*; D058885:Multilocus Sequence Typing; D016133:Polymerase Chain Reaction / Q000379:methods*; D039002:Prophages / Q000235:genetics; D017108:Streptococcus Phages / Q000235:genetics; D013297:Streptococcus pyogenes / Q000145:classification* / Q000235:genetics*; D018089:Superantigens / Q000235:genetics*
chemical_list: D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D002352:Carrier Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D018089:Superantigens; C014689:streptococcal M protein