QUANTITATIVE DETECTION OF BACILLUS ANTHRACIS BY SYBR GREEN I REAL-TIME POLYMERASE CHAIN REACTION / 解放军医学杂志

ABSTRACT

Anthrax is a fatal infectious disease of human and livestock and is caused by Bacillus anthracis. To establish a simple and specific assay for the clinical diagnosis of B. Anthracis infection, two oligonucleotide primers specific for the cap region of plasmid pXO2 and two specific for the pag region of plasmid pXO1 were designed and synthesised for polymerase chain reaction (PCR). The fluorescence dye SYBR Green I incorporated into the double strands was used to quantitate, and the specificity of PCR product was confirmed by the melting curve. The results showed that 0.8?mol/L primers and 3.0mmol/L Mg 2+ were optimal for this quantitative PCR assay. The sensitivity of this assay was 10 3 copies per millitier B. anthracis could be specifically distinguished from other B. cereus group of bacteria. with this assay. SYBR Green I real-time PCR appeared to be a rapid, accurate and specific methad for quantitative analysis of B. anthracis.