基于磁珠的可见光检测微阵列信号的新方法

以磁珠作为标记物,提出了一种在微阵列上检测核酸的新方法。该法基于生物素同链霉亲和素的亲和作用,利用磁珠的超顺磁性和宏观可见特性,使得杂交结果可在普通的光学显微镜或放大镜下检测,甚至肉眼可见。以合成探针为对照,用参比荧光标记染料Cy3标记方法,对这种新方法的检出限进行了研究,并在此基础上采用大肠杆菌16s rDNA的PCR产物作为样品,进行了细菌检测的尝试,取得了较好的实验结果。本法所得的实验结果易于观测,无需采用大型的荧光检测仪器,因而大大降低了检测成本,与其它可见光检测方法(如金胶银染等)相比,具有方便、快捷的特点。这种新方法在传染病检测和环境监测中将具有广阔的应用前景。

A New Visible Light-assisted DNA Array Detection Method with Magnetic Beads

A novel method that utilizes magnetic beads as labeling agent for detecting DNA hybridization on microarrays has been presented.The method is based on the binding affinity between biotinylated oligonucleotides and streptavidin-coated magnetic beads,through which hybridization results can be assessed with a conventional optical microscope,a magnifier or even naked eyes.The stability and detection limit of this method have been studied in comparison with fluorescence labeling method.Attempts to identify bacterium have been performed using E.coli 16s rDNA PCR amplicons as samples,with good results obtained.The scheme significantly facilitates the detection compared with time-consuming colloid gold-silver enhancement method,moreover,it greatly reduces the cost of the detection when compared with common fluorescence labeling techniques because of exemption of cumbersome and expensive scanners.This method has great potential for applications in infectious disease diagnostics and environmental monitoring.