Identification of Capsule-Forming Bacillus Anthracis Spores with the PCR and a Novel Dual-Probe Hybridization Format
Abstract
Anthrax is a fatal infection of humans and livestock that is caused by the gram-positive bacterium Bacillus anthracis. The virulent strains of B. anthracis are encapsulated and toxigenic. In this paper we describe the development of a PCR technique for identifying spores of B. anthracis. Two 20- mer oligonucleotide primers specific for the capB region of 60-MDa plasmid pXO2 were used for amplification. The amplification products were detected by using biotin- and fluorescein-labeled probes in a novel dual-probe hybridization format. Using the combination of PCR amplification and dual-probe hybridization, we detected two copies of the bacterial genome. Because the PCR assay could detect a minimum of 100 unprocessed spores per PCR mixture, we attempted to facilitate the release of DNA by comparing the effect of limited spore germination with the effect of mechanical spore disruption prior to PCR amplification. The two methods were equally effective and allowed us to identify signal spore of B. anthracis in PCR mixtures
Document Details
- Document Type
- Technical Report
- Publication Date
- May 01, 1994
- Accession Number
- ADA285333
Entities
People
- Carl Mitchell
- Malcolm Johns
- Suresh D. Pillai
- Timothy C. Reif
Organizations
- Naval Medical Research Center