Specific detection of bacillus anthracis using a TaqMan mismatch amplification mutation assay

Single nucleotide polymorphisms (SNPs) are increasingly recognized as important diagnostic markers for the detection and differentiation of Bacillus anthracis. The use of SNP markers for identifying B. anthracis DNA in environmental samples containing genetically similar bacteria requires the abilit...

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Publié dans:BioTechniques. - 1993. - 38(2005), 5 vom: 15. Mai, Seite 731-5
Auteur principal: Easterday, William R (Auteur)
Autres auteurs: Van Ert, Matthew N, Zanecki, Shaylan, Keim, Paul
Format: Article
Langue:English
Publié: 2005
Accès à la collection:BioTechniques
Sujets:Evaluation Study Journal Article Technical Report Validation Study Taq Polymerase EC 2.7.7.-
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520 |a Single nucleotide polymorphisms (SNPs) are increasingly recognized as important diagnostic markers for the detection and differentiation of Bacillus anthracis. The use of SNP markers for identifying B. anthracis DNA in environmental samples containing genetically similar bacteria requires the ability to amplify and detect DNA with single nucleotide specificity. We designed a TaqMan mismatch amplification mutation assay (TaqMAMA) around a SNP in the plcR gene of B. anthracis. The assay permits specific, low-level detection (25 fg DNA) of this B. anthracis-specific SNP, even in the presence of environmental DNA extracts containing a 20,000-fold excess of the alternate allele. We anticipate that the ability to selectively amplify and detect low copy number DNAs with single nucleotide specificity will represent a valuable tool in the arena of biodefense and microbial forensics 
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