Abstract:To establish and develop a assay for quick,special and sensitive detection of Bacillus anthracis. By Using Real-time fluorescence polymerase chain reaction based on the TaqMan technology with anti-contamination UDG systems and ROX reference dye,the special probes and primers were designed from pag gene and cap gene on two plasmids, pXO1,pXO2,as well as a ropB gene on chromosome.The amplification efficiency of different company'sreagents were compared.The sensitivity were evaluated by serial dilution of gene colon, internal template and B.anthracis strains.The specifity was confirmed by amolifying real DNAs from bacteria,and the reprentitive strains were identified and assessed by dule-blind.An internal control was added to the reaction mixture in order to detect the presence of PCR inhibitors that were often found in biological samples.Applying the exterior and interior standards, we developed two ways to exactly quantify the samples,and we stimulated infective spleen samples to evaluate the ability for actual application.The results showed that the quantitative real-time PCR assay was sentitive and specific for rapid identification of B.anthracis. It is well suited to identify B.anthracis in case of emergency, bioterrorist attack and surveillance of epidemics.