Indirect detection of Bacillus anthracis using real-time PCR to detect amplified gamma phage DNA.
Typical real-time PCR methods used to identify Bacillus anthracis do not distinguish between viable and non-viable spores, which would be critical in any first response and remediation scenarios. This study combined both real-time PCR, using primers specifically designed for gamma phage, with the highly specific gamma phage amplification into one simple assay to indirectly detect Bacillus anthracis. Since the amplification of gamma phage only occurs in the presence of a suitable host, the detection of increasing concentrations of progeny gamma phage DNA using real-time PCR implies the presence of viable Bacillus anthracis cells. This method detected a starting Bacillus anthracis concentration of 207 cfu/mL, equivalent to less than one cell in 20 microL, in less than 5 h.
Reiman
Robert W
RW
Department of Chemistry and Geochemistry, Colorado School of Mines, Golden, CO, USA. rreiman@mines.edu
Atchley
Daniel H
DH
Voorhees
Kent J
KJ
D017103:Bacillus Phages / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity / Q000502:physiology*; D001408:Bacillus anthracis / Q000145:classification / Q000254:growth & development / Q000302:isolation & purification / Q000821:virology*; D015373:Bacterial Typing Techniques; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013045:Species Specificity
D017931:DNA Primers; D004279:DNA, Viral