Development of a universal probe based microRNA assay and detection of plasma microRNA-146a of human brucellosis / 中国人兽共患病学报

ABSTRACT

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 microRNA146a 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.