一种快速定量检测炭疽杆菌方法的建立  

Establishment of a rapid quantitative method for Bacillus anthracis

作　　者：陈苏红[1] 张敏丽[1] 牟航[1] 管伟[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：《中华检验医学杂志》2003年第2期98-100,共3页Chinese Journal of Laboratory Medicine

摘　　要：目的　建立一种快速、准确、特异定量检测炭疽杆菌的方法。方法　根据复合探针荧光定量分析原理 ,以炭疽杆菌染色体rpoB基因为靶序列 ,设计合成引物和探针 ,对炭疽杆菌进行实时定量聚合酶链反应 (PCR)检测 ,并探讨荧光探针与淬灭探针用量及比例、镁离子浓度、淬灭探针长度对定量结果的影响。结果　本法最适条件 :荧光探针浓度 30 0nmol/L、荧光探针与淬灭探针的比例为1/ 2 ,镁离子浓度为 3mmol/L ,淬灭探针长 15个核苷酸 ,该法检测炭疽杆菌的灵敏度达 10 3 拷贝 ,能特异区分炭疽杆菌与其他蜡样杆菌。结论　复合探针荧光定量PCR技术能够快速准确、特异、敏感地对炭疽杆菌进行定量分析 ,可为临床诊断提供帮助。Objective To establish a rapid specific quantitative assay for Bacillus anthracis detection. Methods According to the principle of complex probe quantitative assay, the primers and quantitative probes targeted at chromatosome DNA rpoB were designed and applied to detect Bacillus anthracis. The influence factor of quantitative PCR were determined. Results The optimal system of this method was aquired: the length of quenching probe is 15mer,the ratio of fluorescent probe to quenching probe is 1/2 and the concentrtion of Mg 2+ is 3 mmol/L.The sensitivity of this assay for Bacillus anthracis is 10 3 copies. It can distinguish Bacillus anthracis from other closely related Bacillus. Conclusion The method can rapidly quantitatively detect the Bacillus anthracis with high sensitivity and specificity, it can be applied to clinical diagnosis.

关 键 词：炭疽芽孢杆菌 聚合酶链反应 分子探针技术 

分 类 号：R446.5[医药卫生—诊断学]