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 single spores of B. anthracis in PCR mixtures.

journal_name

Appl Environ Microbiol

authors

Reif TC,Johns M,Pillai SD,Carl M

doi

10.1128/AEM.60.5.1622-1625.1994

subject

Has Abstract

pub_date

1994-05-01 00:00:00

pages

1622-5

issue

5

eissn

0099-2240

issn

1098-5336

journal_volume

60

pub_type

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