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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| Veröffentlicht in: | BioTechniques 2005-05, Vol.38 (5), p.731-735 |
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| container_title | BioTechniques |
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| creator | EASTERDAY, William R VAN ERT, Matthew N ZANECKI, Shaylan KEIM, Paul |
| description | 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. |
| doi_str_mv | 10.2144/05385ST03 |
| format | Article |
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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. 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Psychology ; In Situ Hybridization - methods ; Molecular and cellular biology ; Polymerase Chain Reaction - methods ; Polymorphism, Single Nucleotide - genetics ; Reproducibility of Results ; Sensitivity and Specificity ; Sequence Alignment - methods ; Sequence Analysis, DNA - methods ; Taq Polymerase - metabolism</subject><ispartof>BioTechniques, 2005-05, Vol.38 (5), p.731-735</ispartof><rights>2005 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=16758891$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/15945372$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>EASTERDAY, William R</creatorcontrib><creatorcontrib>VAN ERT, Matthew N</creatorcontrib><creatorcontrib>ZANECKI, Shaylan</creatorcontrib><creatorcontrib>KEIM, Paul</creatorcontrib><title>Specific detection of Bacillus anthracis using a TaqMan® mismatch amplification mutation assay</title><title>BioTechniques</title><addtitle>Biotechniques</addtitle><description>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.</description><subject>Bacillus anthracis - classification</subject><subject>Bacillus anthracis - genetics</subject><subject>Bacillus anthracis - isolation & purification</subject><subject>Base Pair Mismatch - genetics</subject><subject>Biological and medical sciences</subject><subject>Diverse techniques</subject><subject>DNA Mutational Analysis - methods</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>In Situ Hybridization - methods</subject><subject>Molecular and cellular biology</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Polymorphism, Single Nucleotide - genetics</subject><subject>Reproducibility of Results</subject><subject>Sensitivity and Specificity</subject><subject>Sequence Alignment - methods</subject><subject>Sequence Analysis, DNA - methods</subject><subject>Taq Polymerase - metabolism</subject><issn>0736-6205</issn><issn>1940-9818</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2005</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpF0MlOw0AMANARAtFSOPADaC5wC8y-HKFik4o4tJwjZzJDg7I1kxz6U3wEX0agRZxsS8-WbYTOKblmVIgbIrmRyxXhB2hKrSCJNdQcoinRXCWKETlBJzF-EMKI1vYYTai0QnLNpihdtt4VoXA49713fdHUuAn4DlxRlkPEUPfrbiwiHmJRv2PAK9i8QP31iasiVtC7NYaqLX9GwG93NfS7BGKE7Sk6ClBGf7aPM_T2cL-aPyWL18fn-e0iaRm3feKsos4GQpVlilOpFLM898SBASKCzp0QWgQ6CiOFDiJ3eZ5lgjHJFc8Un6Gr3dy2azaDj306rud8WULtmyGmSlvGKDUjvNjDIat8nrZdUUG3Tf9eMoLLPYDooAwd1OP5_05paYyl_Bs6Fm_q</recordid><startdate>20050501</startdate><enddate>20050501</enddate><creator>EASTERDAY, William R</creator><creator>VAN ERT, Matthew N</creator><creator>ZANECKI, Shaylan</creator><creator>KEIM, Paul</creator><general>Eaton</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope></search><sort><creationdate>20050501</creationdate><title>Specific detection of Bacillus anthracis using a TaqMan® mismatch amplification mutation assay</title><author>EASTERDAY, William R ; VAN ERT, Matthew N ; ZANECKI, Shaylan ; KEIM, Paul</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p239t-c961c9f01692631566293de0ca8a04f7dc4474f1f018547f4dcddbb4225363b63</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2005</creationdate><topic>Bacillus anthracis - classification</topic><topic>Bacillus anthracis - genetics</topic><topic>Bacillus anthracis - isolation & purification</topic><topic>Base Pair Mismatch - genetics</topic><topic>Biological and medical sciences</topic><topic>Diverse techniques</topic><topic>DNA Mutational Analysis - methods</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>In Situ Hybridization - methods</topic><topic>Molecular and cellular biology</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Polymorphism, Single Nucleotide - genetics</topic><topic>Reproducibility of Results</topic><topic>Sensitivity and Specificity</topic><topic>Sequence Alignment - methods</topic><topic>Sequence Analysis, DNA - methods</topic><topic>Taq Polymerase - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>EASTERDAY, William R</creatorcontrib><creatorcontrib>VAN ERT, Matthew N</creatorcontrib><creatorcontrib>ZANECKI, Shaylan</creatorcontrib><creatorcontrib>KEIM, Paul</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><jtitle>BioTechniques</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>EASTERDAY, William R</au><au>VAN ERT, Matthew N</au><au>ZANECKI, Shaylan</au><au>KEIM, Paul</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Specific detection of Bacillus anthracis using a TaqMan® mismatch amplification mutation assay</atitle><jtitle>BioTechniques</jtitle><addtitle>Biotechniques</addtitle><date>2005-05-01</date><risdate>2005</risdate><volume>38</volume><issue>5</issue><spage>731</spage><epage>735</epage><pages>731-735</pages><issn>0736-6205</issn><eissn>1940-9818</eissn><coden>BTNQDO</coden><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.</abstract><cop>Natick, MA</cop><pub>Eaton</pub><pmid>15945372</pmid><doi>10.2144/05385ST03</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| source | Taylor & Francis Open Access; MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals |
| subjects | Bacillus anthracis - classification Bacillus anthracis - genetics Bacillus anthracis - isolation & purification Base Pair Mismatch - genetics Biological and medical sciences Diverse techniques DNA Mutational Analysis - methods Fundamental and applied biological sciences. Psychology In Situ Hybridization - methods Molecular and cellular biology Polymerase Chain Reaction - methods Polymorphism, Single Nucleotide - genetics Reproducibility of Results Sensitivity and Specificity Sequence Alignment - methods Sequence Analysis, DNA - methods Taq Polymerase - metabolism |
| title | Specific detection of Bacillus anthracis using a TaqMan® mismatch amplification mutation assay |
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