Rapid and effective detection of anthrax spores in soil by PCR.
OBJECTIVE
To detect Bacillus anthracis DNA from soil using rapid and simple procedures.


RESULTS
Various amounts of B. anthracis Pasteur II spores were added artificially to 1 g of soil, which was then washed with ethanol and sterile water. Enrichment of the samples in trypticase soy broth was performed twice. A DNA template was prepared from the second enrichment culture using a FastPrep instrument. The template was then used for nested and real-time polymerase chain reaction (PCR) with B. anthracis-specific primers, to confirm the presence of B. anthracis chromosomal DNA and the pXO1/pXO2 plasmids.


CONCLUSIONS
One cell of B. anthracis in 1 g of soil could be detected by nested and real-time PCR. The usefulness of the PCR method using field samples was also confirmed.


CONCLUSIONS
The results indicate that this could be a useful method for detecting anthrax-spore contaminated soil with high sensitivity. Its application could have great impact on the progress of epidemiological surveillance.
Cheun
H I
HI
Department of Applied Veterinary Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, Japan.
Makino
S-I
SI
Watarai
M
M
Erdenebaatar
J
J
Kawamoto
K
K
Uchida
I
I
D001408:Bacillus anthracis / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D001483:Base Sequence; D003470:Culture Media; D004269:DNA, Bacterial / Q000032:analysis*; D010957:Plasmids / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D012988:Soil Microbiology; D013171:Spores, Bacterial / Q000235:genetics / Q000302:isolation & purification
D003470:Culture Media; D004269:DNA, Bacterial