Real-time PCR system targeting a chromosomal marker specific for Bacillus anthracis

被引：57

作者：

Antwerpen, Markus H. ^{[1
]}

Zimmermann, Pia ^{[1
]}

Bewley, Kevin ^{[2
]}

Frangoulidis, Dimitrios ^{[1
]}

Meyer, Hermann ^{[1
]}

机构：

[1] Bundeswehr Inst Microbiol, D-80937 Munich, Germany

[2] Hlth Protect Agcy, Salisbury, Wilts, England

来源：

MOLECULAR AND CELLULAR PROBES | 2008年 / 22卷 / 5-6期

关键词：

Bacillus anthracis; Real-time PCR; Chromosomal marker; dhp61; BA5345;

D O I：

10.1016/j.mcp.2008.06.001

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Specific identification of Bacillus anthracis and differentiation from closely related Bacillus cereus and Bacillus thuringiensis strains is still a major diagnostic problem. Commercially available diagnostic kits targeting plasmid-markers cannot differentiate between B. anthracis, non-anthracis Bacillus species harbouring anthrax-specific virulence plasmids, and plasmidless B. anthracis strains. A TaqMan (R) PCR assay was designed targeting sequences of gene locus BA_5345 of the B. anthracis strain Ames. Specificity was determined by using a panel of 328 Bacillus strains; sensitivity was determined by probit analysis. All B. anthracis isolates (n = 92) were specifically detected by using the genomic TaqMan (R) PCR assay whereas 236 strains belonging to 19 Bacillus species other than B. anthracis were PCR negative. The detection limit was determined to be 12.7 copies per reaction (95% confidence interval 10.2-17.5 copies). Here we present an extensively evaluated and - to our current knowledge - specific TaqMan (R) PCR assay for the detection of B. anthracis based on a chromosomal marker. (C) 2008 Elsevier Ltd. All rights reserved.

引用

页码：313 / 315

页数：3

共 23 条

[1] Strategy for identification of Bacillus cereus and Bacillus thuringiensis strains closely related to Bacillus anthracis [J].

Daffonchio, D ;

Raddadi, N ;

Merabishvili, M ;

Cherif, A ;

Carmagnola, L ;

Brusetti, L ;

Rizzi, A ;

Chanishvili, N ;

Visca, P ;

Sharp, R ;

Borin, S .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2006, 72 (02) :1295-1301

[2] TaqMan 5′-nuclease human immunodeficiency virus type IPCR assay with phage-packaged competitive internal control for high-throughput blood donor screening [J].

Drosten, C ;

Seifried, E ;

Roth, WK .

JOURNAL OF CLINICAL MICROBIOLOGY, 2001, 39 (12) :4302-4308

[3] Use of single nucleotide polymorphisms in the plcR gene for specific identification of Bacillus anthracis [J].

Easterday, WR ;

Van Ert, MN ;

Simonson, TS ;

Wagner, DM ;

Kenefic, LJ ;

Allender, CJ ;

Keim, P .

JOURNAL OF CLINICAL MICROBIOLOGY, 2005, 43 (04) :1995-1997

[4] Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis -: One species on the basis of genetic evidence [J].

Helgason, E ;

Okstad, OA ;

Caugant, DA ;

Johansen, HA ;

Fouet, A ;

Mock, M ;

Hegna, I ;

Kolsto, AB .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2000, 66 (06) :2627-2630

[5] 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

[6] Characterization of Bacillus cereus isolates associated with fatal pneumonias:: Strains are closely related to Bacillus anthracis and Harbor B-anthracis virulence genes [J].

Hoffmaster, Alex R. ;

Hill, Karen K. ;

Gee, Jay E. ;

Marston, Chung K. ;

De, Barun K. ;

Popovic, Tanja ;

Sue, David ;

Wilkins, Patricia P. ;

Avashia, Swati B. ;

Drumgoole, Rahsaan ;

Helma, Charles H. ;

Ticknor, Lawrence O. ;

Okinaka, Richard T. ;

Jackson, Paul J. .

JOURNAL OF CLINICAL MICROBIOLOGY, 2006, 44 (09) :3352-3360

[7] Identification of anthrax toxin genes in a Bacillus cereus associated with an illness resembling inhalation anthrax [J].

Hoffmaster, AR ;

Ravel, J ;

Rasko, DA ;

Chapman, GD ;

Chute, MD ;

Marston, CK ;

De, BK ;

Sacchi, CT ;

Fitzgerald, C ;

Mayer, LW ;

Maiden, MCJ ;

Priest, FG ;

Barker, M ;

Jiang, LX ;

Cer, RZ ;

Rilstone, J ;

Peterson, SN ;

Weyant, RS ;

Galloway, DR ;

Read, TD ;

Popovic, T ;

Fraser, CM .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (22) :8449-8454

[8] Evaluation and validation of a real time polymerase chain reaction assay for rapid identification of Bacillus anthracis [J].

Hoffmaster, AR ;

Meyer, RF ;

Bowen, MP ;

Marston, CK ;

Weyant, RS ;

Barnett, GA ;

Sejvar, JJ ;

Jernigan, JA ;

Perkins, BA ;

Popovic, T .

EMERGING INFECTIOUS DISEASES, 2002, 8 (10) :1178-1182

[9] 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

[10] Multiple-locus variable-number tandem repeat analysis reveals genetic relationships within Bacillus anthracis [J].

Keim, P ;

Price, LB ;

Klevytska, AM ;

Smith, KL ;

Schupp, JM ;

Okinaka, R ;

Jackson, PJ ;

Hugh-Jones, ME .

JOURNAL OF BACTERIOLOGY, 2000, 182 (10) :2928-2936

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