Open AccessVir typing: a long-PCR typing method for group A streptococci.
Author(s) -
Don L. Gardiner,
Jon Hartas,
Bart J. Currie,
John D. Mathews,
Darrell J. Kemp,
Kadaba S. Sriprakash
Publication year - 1995
Publication title -
genome research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.556
H-Index - 297
eISSN - 1549-5469
pISSN - 1088-9051
DOI - 10.1101/gr.4.5.288
Subject(s) - biology , typing , haeiii , ethidium bromide , restriction enzyme , dna , microbiology and biotechnology , rapd , agarose gel electrophoresis , genetics , dna extraction , polymerase chain reaction , restriction fragment length polymorphism , gene , population , demography , sociology , genetic diversity
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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