Methods and Compositions
for Detecting Bacillus Species

Case Number CH-036

Inventors: Terrance Leighton, PhD, et al.

Patent No. Title Issue Date Country
Pending Methods and Compositions for Detecting Bacillus Species   United States

Abstract
This invention provides methods and compositions for rapid, sensitive, and highly specific nucleic acid-based detection of B. anthracis in a sample.

Anthrax infections are initiated by endospores of B. anthracis, a Gram-positive soil organism.  Anthrax endospores do not divide, have no measurable metabolism, and are resistant to drying, heat, ultraviolet light, gamma radiation, and many disinfectants.  Endospores introduced into the body by abrasion, inhalation, or ingestion are phagocytosed by macrophages and carried into regional lymph nodes where they enter the blood stream, causing massive septicemia.  Fully virulent or vaccine strains of B. anthracis can be easily identified by their virulence plasmid composition, however, many B. anthracis strains do not contain these plasmids, and, as such, cannot be detected by current methods.  There is currently a need for reliable and rapid detection methods for anthrax bacteria.

Advantage
This method involves detecting within the sspE gene of B. anthracis the presence of a fragment of six contiguous nucleotides of unique sequence that is not present in the sspE genes and other Bacillus bacteria or other non-Bacillus bacteria.  In many embodiments, the methods involve amplifying a nucleic acid comprising at least a fragment of an sspE gene from a sample, and detecting the presence of the unique six contiguous nucleotide fragments in the nucleic acid. 

The invention also provides primers and kits for detection of B. anthracis in a sample, as well as primers and methods for discriminating B. anthracis and near-neighbor chromosomes from each other and those of phylogenetically unrelated bacteria. 

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