Vir typing: a long-PCR typing method for group A streptococci.
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.
Gardiner
D
D
Menzies School of Health Research, Darwin, NT, Australia.
Hartas
J
J
Currie
B
B
Mathews
J D
JD
Kemp
D J
DJ
Sriprakash
K S
KS
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
D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D037521:Virulence Factors; D004706:Endodeoxyribonucleases