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

被引：27

作者：

Easterday, WR

Van Ert, MN

Zanecki, S

Keim, P ^{[1
]}

机构：

[1] No Arizona Univ, Dept Sci Biol, Flagstaff, AZ 86011 USA

[2] TGen, Phoenix, AZ USA

来源：

BIOTECHNIQUES | 2005年 / 38卷 / 05期

关键词：

D O I：

10.2144/05385ST03

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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 (R) 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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收藏

页码：731 / 735

页数：7

相关论文

共 11 条

[1]

Cha R S, 1992, PCR Methods Appl, V2, P14

[2]

EASTERDAY WR, IN PRESS J CLIN MICR

[3] A novel assay for allelic discrimination that combines the fluorogenic 5′ nuclease polymerase chain reaction (TaqMan®) and mismatch amplification mutation assay [J].

Glaab, WE ;

Skopek, TR .

MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS, 1999, 430 (01) :1-12

[4] Fluorescent amplified fragment length polymorphism analysis of Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis isolates [J].

Hill, KK ;

Ticknor, LO ;

Okinaka, RT ;

Asay, M ;

Blair, H ;

Bliss, KA ;

Laker, M ;

Pardington, PE ;

Richardson, AP ;

Tonks, M ;

Beecher, DJ ;

Kemp, JD ;

Kolsto, AB ;

Wong, ACL ;

Keim, P ;

Jackson, PJ .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2004, 70 (02) :1068-1080

[5] Detection of the Bacillus anthracis gyrA gene by using a minor groove binder probe [J].

Hurtle, W ;

Bode, E ;

Kulesh, DA ;

Kaplan, RS ;

Garrison, J ;

Bridge, D ;

House, M ;

Frye, MS ;

Loveless, B ;

Norwood, D .

JOURNAL OF CLINICAL MICROBIOLOGY, 2004, 42 (01) :179-185

[6] Anthrax molecular epidemiology and forensics: using the appropriate marker for different evolutionary scales [J].

Keim, Paul ;

Van Ert, Matthew N. ;

Pearson, Talima ;

Vogler, Amy J. ;

Huynh, Lynn Y. ;

Wagner, David M. .

INFECTION GENETICS AND EVOLUTION, 2004, 4 (03) :205-213

[7] Relief of amplification inhibition in PCR with bovine serum albumin or T4 gene 32 protein [J].

Kreader, CA .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1996, 62 (03) :1102-1106

[8] The incompatibility between the PlcR- and AtxA-controlled regulons may have selected a nonsense mutation in Bacillus anthracis [J].

Mignot, T ;

Mock, M ;

Robichon, D ;

Landier, A ;

Lereclus, D ;

Fouet, A .

MOLECULAR MICROBIOLOGY, 2001, 42 (05) :1189-1198

[9] Phylogenetic discovery bias in Bacillus anthracis using single-nucleotide polymorphisms from whole-genome sequencing [J].

Pearson, T ;

Busch, JD ;

Ravel, J ;

Read, TD ;

Rhoton, SD ;

U'Ren, JM ;

Simonson, TS ;

Kachur, SM ;

Leadem, RR ;

Cardon, ML ;

Van Ert, MN ;

Huynh, LY ;

Fraser, CM ;

Keim, P .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (37) :13536-13541

[10] Genetic diversity in the protective antigen gene of Bacillus anthracis [J].

Price, LB ;

Hugh-Jones, M ;

Jackson, PJ ;

Keim, P .

JOURNAL OF BACTERIOLOGY, 1999, 181 (08) :2358-2362