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Rapid and effective detection of anthrax spores in soil by PCR
Author(s) -
Cheun H.I.,
Makino S.I.,
Watarai M.,
Erdenebaatar J.,
Kawamoto K.,
Uchida I.
Publication year - 2003
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1046/j.1365-2672.2003.02038.x
Subject(s) - bacillus anthracis , spore , polymerase chain reaction , microbiology and biotechnology , biology , plasmid , anthrax toxin , nested polymerase chain reaction , dna , real time polymerase chain reaction , bacteria , gene , genetics , recombinant dna , fusion protein
Aims: To detect Bacillus anthracis DNA from soil using rapid and simple procedures. Methods and 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. Significance and Impact of the Study: 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.