Evaluation and Validation of a Real-Time Polymerase Chain Reaction Assay for Rapid Identification of<i>Bacillus anthracis</i>

Hoffmaster, Alex R.; Meyer, Richard F.; Bowen, Michael P.; Marston, Chung K.; Weyant, Robbin S.; Thurman, Kathy; Messenger, Sharon; Minor, Erin E.; Winchell, Jonas M.; Rasmussen, Max V.; Newton, Bruce R.; Parker, Jake; Morrill, William E.; McKinney, Nancy; Barnett, Gwen A.; Sejvar, James J.; Jernigan, John A.; Perkins, Bradley A.; Popović, Tanja

doi:10.3201/eid0810.020393

Cited by 119 publications

(99 citation statements)

References 23 publications

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“…Polymerase chain reaction (PCR) was used during the 2001 anthrax attacks, with primer and probe sets targeting each of the plasmids as well as the chromosome. 39 Alam et al 40 improved PCR sensitivity by using 2 signatures for the gene coding for edema factor; Christensen et al 41 determined that real-time PCR could identify 50 fg, or 9 genome equivalents of B. anthracis Ames using either the RAPID or Smart Cycler platforms. However, PCR requires clean samples in a small volume and is not generally suitable for field use.…”

Section: Discussionmentioning

confidence: 99%

Comprehensive Laboratory Evaluation of a Highly Specific Lateral Flow Assay for the Presumptive Identification ofBacillus anthracisSpores in Suspicious White Powders and Environmental Samples

et al. 2016

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We conducted a comprehensive, multiphase laboratory evaluation of the Anthrax BioThreat Alert Ò test strip, a lateral flow immunoassay (LFA) for the rapid detection of Bacillus anthracis spores. The study, conducted at 2 sites, evaluated this assay for the detection of spores from the Ames and Sterne strains of B. anthracis, as well as those from an additional 22 strains. Phylogenetic near neighbors, environmental background organisms, white powders, and environmental samples were also tested. The Anthrax LFA demonstrated a limit of detection of about 10 6 spores/mL (ca. 1.5 · 10 5 spores/assay). In this study, overall sensitivity of the LFA was 99.3%, and the specificity was 98.6%. The results indicated that the specificity, sensitivity, limit of detection, dynamic range, and repeatability of the assay support its use in the field for the purpose of qualitatively evaluating suspicious white powders and environmental samples for the presumptive presence of B. anthracis spores.

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Section: Discussionmentioning

confidence: 99%

Comprehensive Laboratory Evaluation of a Highly Specific Lateral Flow Assay for the Presumptive Identification ofBacillus anthracisSpores in Suspicious White Powders and Environmental Samples

et al. 2016

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“…[21,27,43]. PCR is the most widely used molecular diagnostic technique due to its fast and easy to use protocol.…”

Section: Pcr Implementation In Diagnosticsmentioning

confidence: 99%

Potentials and limitations of molecular diagnostic methods in food safety

Lauri

Mariani

2008

Genes Nutr

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Molecular methods allow the detection of pathogen nucleic acids (DNA and RNA) and, therefore, the detection of contamination in food is carried out with high selectivity and rapidity. In the last 2 decades molecular methods have accompanied traditional diagnostic methods in routine pathogen detection, and might replace them in the upcoming future. In this review the implementation in diagnostics of four of the most used molecular techniques (PCR, NASBA, microarray, LDR) are described and compared, highlighting advantages and limitations of each of them. Drawbacks of molecular methods with regard to traditional ones and the difficulties encountered in pathogen detection from food or clinical specimen are also discussed. Moreover, criteria for the choice of the target sequence for a secure detection and classification of pathogens and possible developments in molecular diagnostics are also proposed.

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“…30 These genetic markers provide limited specificity and require additional timeconsuming and labor-intensive post-PCR analysis steps. Other areas of the chromosome have also been investigated as potential DNA-targets for identification purposes, including the so-called BA813 [31][32][33][34][35][36][37][38] and BA5510 sequences, 19 genes bclB, 39 sap, 40,41 saspB, 5,42 and sspE, 22,43 the B-type small acid-soluble spore protein gene (SASP), 44 a glycosyltransferase group 1 family protein, 45 a protein showing similarities with an abhydrolase, 18 and several DNA loci located on prophage regions, 17 i.e., BA5345, 21 BA5357, 46 and PL3. 47 Although most of these regions have been claimed to be anthrax-specific, B. cereus strains sometimes yield false positive results.…”

Section: Literature Survey Of Pcr-based Detection Methodsmentioning

confidence: 99%

In silico and in vitro evaluation of PCR-based assays for the detection ofBacillus anthracischromosomal signature sequences

et al. 2013

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Evaluation and Validation of a Real-Time Polymerase Chain Reaction Assay for Rapid Identification ofBacillus anthracis

Cited by 119 publications

References 23 publications

Comprehensive Laboratory Evaluation of a Highly Specific Lateral Flow Assay for the Presumptive Identification ofBacillus anthracisSpores in Suspicious White Powders and Environmental Samples

Comprehensive Laboratory Evaluation of a Highly Specific Lateral Flow Assay for the Presumptive Identification ofBacillus anthracisSpores in Suspicious White Powders and Environmental Samples

Potentials and limitations of molecular diagnostic methods in food safety

In silico and in vitro evaluation of PCR-based assays for the detection ofBacillus anthracischromosomal signature sequences

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