Generation of a specific marker to discriminate Gacillus anthracis from other bacteria of the Bacillus cereus group.
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.
Division of Molecular and Life Sciences, Hanyang University, Ansan 426-791, Korea.
Tai Hyoung
TH
Kim
Gwi Moon
GM
Seo
Kyoung Hwa
KH
Jung
Seong Joo
SJ
Kim
Jee Cheon
JC
Kim
Kwang Geun
KG
Oh
Bon-Sung
BS
Koo
Young Gyu
YG
Chai
D001408:Bacillus anthracis / Q000145:classification* / Q000235:genetics; D001409:Bacillus cereus / Q000145:classification / Q000235:genetics; D001413:Bacillus thuringiensis / Q000145:classification / Q000235:genetics; D001483:Base Sequence; D015415:Biomarkers / Q000032:analysis*; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D019105:Random Amplified Polymorphic DNA Technique; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D015415:Biomarkers; D017931:DNA Primers; D004269:DNA, Bacterial