D Gardiner, J Hartas, B Currie, J D Mathews, D J Kemp, K S Sriprakash
{"title":"Vir typing: a long-PCR typing method for group A streptococci.","authors":"D Gardiner, J Hartas, B Currie, J D Mathews, D J Kemp, K S Sriprakash","doi":"10.1101/gr.4.5.288","DOIUrl":null,"url":null,"abstract":"<p><p>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.</p>","PeriodicalId":77315,"journal":{"name":"PCR methods and applications","volume":"4 5","pages":"288-93"},"PeriodicalIF":0.0000,"publicationDate":"1995-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"88","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"PCR methods and applications","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1101/gr.4.5.288","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 88
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.