实时荧光定量PCR法检测炭疽杆菌  

QUANTITATIVE DETECTION OF BACILLUS ANTHRACIS BY SYBR GREEN I REAL-TIME POLYMERASE CHAIN REACTION

作　　者：陈苏红[1] 张敏丽[1] 牟航[1] 管伟[1] 李鲁[1] 王升启[1] 

机构地区：[1]军事医学科学院放射医学研究所,北京100850

出　　处：《解放军医学杂志》2003年第3期268-270,共3页Medical Journal of Chinese People's Liberation Army

摘　　要：为建立一种简便而特异的炭疽杆菌检测方法用作临床诊断工具 ,以pXO1质粒上的 pag基因及pXO2质粒上的cap基因为靶合成特异引物 ,用DNA嵌入荧光染料SYBRGreenI进行定量PCR扩增 ,在PCR后进行熔解曲线分析以确定得到的产物是否为目的产物。定量PCR条件优化结果为 :引物浓度 0 8μmol/L、Mg2 +5 0mmol/L。该法检测炭疽的灵敏度达 10 3拷贝 ,并能区分炭疽杆菌与其他蜡样杆菌。表明实时荧光定量PCR技术能够快速准确。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 0.8μmol/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.

关 键 词：炭疽杆菌 聚合酶链反应 定量 实时检测 

分 类 号：R378.72[医药卫生—病原生物学]