摘要
目的建立一种通用、敏感、特异的microRNA检测方法,用于检测布鲁氏菌病患者血浆microRNA-146a的表达,探讨其作为诊断标识的可能性。方法以microRNA-146a模拟物为待检物,通过正交实验设计,对退火温度、探针浓度、试剂盒选择进行优化,建立特异敏感的通用检测方法;从布病患者和正常健康人血浆样本提取RNA,应用通用探针法进行检测,比较布病患者血浆microRNA-146a的表达变化。结果建立了优化的通用探针法,与染料法相比特异性更强、扩增范围更大,具有通用检测能力。应用通用探针法检测布鲁氏菌病患者血浆microRNA-146a,发现布病患者血浆中microRNA-146a的表达被抑制(P<0.01),表明其可能作为布病诊断的分子标识。结论经优化的通用探针法是一种通用、敏感、特异的检测方法,可用于microRNA的检测,microRNA-146a可能作为布病诊断的标识分子。
We developed a universal probe based microRNA detection assay and applied it to detect microRNA-146a in human brucellosis, testing the possibility of using it as diagnosis signature. By using orthogonal design, the annealing temperature, probe concentration and commercial kits were optimized and the assay was developed. Total RNAs were isolated from plasma of human brucellosis and healthy control, and microRNA-146a was detected and compared. Results reveal that the optimized universal probe assay was established, which was more specific than the SYBR GreenI assay, and had a wider range of amplification. Compared with healthy control, the application of universal probe assay for the detection of serum microRNA-146a in patients with brucellosis was significantly inhibited （P〈0.01）. Implying the potential of microRNA-146a as biomarker in diagnosis of brucellosis.It is suggested that universal probe based assay is a universal, specific and sensitive method for microRNA detection. MicroRNA-146a represents a potential biomarker for human brucellosis diagnosis.
作者
程慧敏
魏青青
张欢
刘金玲
韩小虎
王大力
王兴龙
CHENG Hui-min WEI Qing-qing ZHANG Huan LIU Jin-ling HAN Xiao-hu WANG Da-li WANG Xing-long(Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun 130122, China Shenyang Agriculture University, Shenyang 110866, China Pestis and Brucellosis Therapeutic Base, Chinese Centers for Disease Control and Prevention, Baicheng 137000, China)
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2017年第5期441-448,共8页
Chinese Journal of Zoonoses
