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J Microbiol Biotechnol ; 17(5): 806-11, 2007 May.
Artigo em Inglês | MEDLINE | ID: mdl-18051303

RESUMO

Bacillus anthracis is a soil pathogen capable of causing anthrax that is closely related to several environmental species, including B. cereus, B. mycoides, and B. thuringiensis. DNA homology studies showed that B. anthracis, B. cereus, B. mycoides, and B. thuringiensis are closely related, with a high sequence homology. To establish a method to specifically detect B. anthracis in situations such as environmental contamination, we initially performed RAPD-PCR with a 10-mer random primer and confirmed the presence of specific PCR bands only in B. anthracis species. One region specific for B. anthracis was cloned and sequenced, and an internal primer set was designed to amplify a 241-bp DNA fragment within the sequenced region. The PCR system involving these specific primer sets has practical applications. Using lyses methods to prepare the samples for PCR, it was possible to quickly amplify the 241-bp DNA segment from samples containing only a few bacteria. Thus, the PCR detection method developed in this study is expected to facilitate the monitoring of environmental B. anthracis contamination.


Assuntos
Bacillus anthracis/classificação , Biomarcadores/análise , DNA Bacteriano/genética , Reação em Cadeia da Polimerase/métodos , Bacillus anthracis/genética , Bacillus cereus/classificação , Bacillus cereus/genética , Bacillus thuringiensis/classificação , Bacillus thuringiensis/genética , Sequência de Bases , Clonagem Molecular , Primers do DNA/genética , DNA Bacteriano/química , Dados de Sequência Molecular , Técnica de Amplificação ao Acaso de DNA Polimórfico , Sensibilidade e Especificidade , Análise de Sequência de DNA
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