Vir typing: a long-PCR typing method for group A streptococci.

  1. D Gardiner,
  2. J Hartas,
  3. B Currie,
  4. J D Mathews,
  5. D J Kemp, and
  6. K S Sriprakash
  1. Menzies School of Health Research, Darwin, NT, Australia.

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.

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