微滴数字PCR定量检测全血样品中结核分枝杆菌特异CFP10基因

被引：7

作者：

宋能 ^{[1
]}

谭杨 ^{[1
]}

罗凤玲 ^{[1
]}

关清 ^{[1
]}

闫明哲 ^{[2
]}

潘勤 ^{[1
]}

章晓联 ^{[1
]}

机构：

[1] 武汉大学病毒学国家重点实验室/湖北省过敏及免疫相关疾病重点实验室/武汉大学医学研究院/武汉大学基础医学院免疫学系

[2] 武汉市医疗救治中心

来源：

中国生化药物杂志 | 2016年 / 36卷 / 02期

基金：

国家杰出青年科学基金;

关键词：

微滴数字PCR; 结核分枝杆菌; 定量检测; CFP10;

D O I：

暂无

中图分类号：

R52 [结核病]; R440 [];

学科分类号：

摘要：

目的采用微滴数字PCR技术(droplet digital PCR,dd PCR)检测全血中结核分枝杆菌特异性CFP10(10-k Da culture filtrate protein,CFP10)基因的拷贝数含量。方法收集10例活动性肺结核患者(active tuberculosis,a TB)、10例肺外结核患者(extrapulmonary tuberculosis,EPTB)和5例健康对照者(healthy donars,HDs)全血并提取DNA,优化引物最佳扩增条件并制定重组质粒标准曲线,对样本中CFP10基因的拷贝数含量分别用dd PCR与实时定量PCR(quantitative real-time PCR,q PCR)进行检测和比较。结果对重组质粒的灵敏度检测显示dd PCR技术明显优于q PCR技术。对样本的检测显示q PCR技术检测a TB、EPTB患者与健康人的全血中CFP10含量之间无统计学意义;但是dd PCR技术检测a TB、EPTB患者与健康人之间有显著统计学意义(P<0.0001);dd PCR检测a TB和EPTB患者(共20例)CFP10基因绝对定量的浓度约为3.4～94.0 copies/μL。结论本研究首次报道采用dd PCR技术能灵敏地用于TB患者全血的诊断。

引用

页码：10 / 15

页数：6

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