Specific detection of Bacillus anthracis using a TaqMan® mismatch amplification mutation assay

William R. Easterday, Matthew N. Van Ert, Shaylan Zanecki, Paul Keim

Research output: Contribution to journalArticle

31 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)731-735
Number of pages5
JournalBioTechniques
Volume38
Issue number5
DOIs
StatePublished - May 2005

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

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