ISOLATION OF A SPECIFIC CHROMOSOMIC DNA-SEQUENCE OF BACILLUS-ANTHRACIS AND ITS POSSIBLE USE IN DIAGNOSIS

A 277-bp long DNA fragment, Ba813, was isolated from an avirulent Baci llus anthracis strain 7700 genomic library. Two oligonucleotides deriv ed from the Ba813 sequence were used as primers in polymerase chain re action tests on genomic DNA from 28 Bacillus anthracis and from 33 het erologous bacteria strains. A specific, 152-bp long DNA fragment was a mplified only when Bacillus anthracis DNA was used as the target. The amplified product was analysed by non-radioactive sandwich hybridisati on in microtiter plates using two oligonucleotides. The capture oligon ucleotide CI was covalently linked onto aminated wells of microtiter p lates. The detection oligonucleotide D3 was labelled with biotine. The hybrid molecules were detected by avidine conjugated with alkaline ph osphatase and chromogenic substrate. Amplification of Ba813 sequence m ay provide the basis for rapid and reliable assay for the detection an d identification of Bacillus anthracis.