title: Vir typing: a long-PCR typing method for group A streptococci.
abstract: We have developed a new procedure (Vir typing) for typing Streptococcus pyogenes, by amplifying the entire 5- to 7-kb variable vir regulon by long PCR. The amplified DNA is then cleaved with HaeIII and visualized by ethidium bromide fluorescence after agarose gel electrophoresis. A simple procedure for preparing DNA of sufficiently high quality from 96 samples was employed simultaneously. This DNA was also used to develop a random amplified polymorphic DNA (RAPD) procedure. The discriminatory power of the two DNA-based procedures was compared with previous methods, M typing, and multilocus enzyme electrophoresis. Both procedures were highly discriminatory, but the stoichiometric yield of restriction fragments in Vir typing allows unambiguous interpretation of results.
authors.affiliation: Menzies School of Health Research, Darwin, NT, Australia.
authors.forename: D
authors.initials: D
authors.lastname: Gardiner
authors.forename: J
authors.initials: J
authors.lastname: Hartas
authors.forename: B
authors.initials: B
authors.lastname: Currie
authors.forename: J D
authors.initials: JD
authors.lastname: Mathews
authors.forename: D J
authors.initials: DJ
authors.lastname: Kemp
authors.forename: K S
authors.initials: KS
authors.lastname: Sriprakash
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004706:Endodeoxyribonucleases / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D018085:Regulon; D015183:Restriction Mapping; D012680:Sensitivity and Specificity; D013290:Streptococcal Infections / Q000382:microbiology* / Q000635:transmission; D013297:Streptococcus pyogenes / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D037521:Virulence Factors
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D037521:Virulence Factors; D004706:Endodeoxyribonucleases