Rapid and effective detection of anthrax spores in soil by PCR

被引:35
作者
Cheun, HI
Makino, SI
Watarai, M
Erdenebaatar, J
Kawamoto, K
Uchida, I
机构
[1] Obihiro Univ Agr & Vet Med, Dept Appl Vet Sci, Obihiro, Hokkaido 0808555, Japan
[2] Obihiro Univ Agr & Vet Med, Res Ctr Anim Hyg & Food Safety, Obihiro, Hokkaido 0808555, Japan
[3] Natl Inst Anim Hlth, Hokkaido Res Stn, Sapporo, Hokkaido, Japan
[4] Inst Vet Med, Immunol Res Ctr, Lab Bacteriol & Infect Dis, Ulaanbaatar, Mongolia
关键词
Bacillus anthracis; detection; nested PCR; real-time PCR; soil;
D O I
10.1046/j.1365-2672.2003.02038.x
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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.
引用
收藏
页码:728 / 733
页数:6
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