基于生物信号纳米放大技术的戊肝病毒检测基因芯片  

Visual Gene Detection Chip for HEV Based on Bio-Signal with Nano-Amplification Technology

作　　者：杨永[1] 翟建新[1] 王业富[1] 

机构地区：[1]武汉大学生命科学学院,湖北武汉430072

出　　处：《武汉大学学报（理学版）》2007年第4期475-480,共6页Journal of Wuhan University:Natural Science Edition

基　　金：湖北省自然科学基金资助项目(2001AA305B01)

摘　　要：建立了一种利用"双探针夹心银染"技术快速检测戊型肝炎病毒(HEV)的基因芯片方法.提取的病毒核酸经过RT-PCR扩增,PCR体系中用掺入生物素修饰的dUTP扩增得到带有生物素的核酸片段,5′末端-NH2修饰的同源寡核苷酸作为捕获探针固定到处理后的玻片上,该探针识别病毒的带有生物素的核酸片段,而链霉亲和素标记的胶体金与生物素紧密结合,通过银沉积在金颗粒表面放大信号可以目视结果.采用该方法对已经确证的86个血清样本进行检测,检出52个阳性和34个阴性样本,结果与荧光定量PCR检出一致.A rapid detection of hepatitis E virus （HEV） gene chip with nano-amplification technique was developed. The whole frame can be described with ＂sandwich hybridization of double probes coupled with silver stain enhancement＂. To detect Hepatitis E Virus, the virus RNA was extracted from the patient serum, then followed by RT-PCR. 5＇-end-NH2 modified oligonucleotides were designed to fabricate a DNA microarry in order to discriminate the target biotinylated DNA amplified by RT-PCR, the whole reaction system with partial dTTP was replaced by bio-dUTP, the signals generated with this method results from the precipitation of silver onto nanogold particles bound to streptavidin. The signals can be seen with naked eyes, which reduces the cost of detection. A total of 86 sera were tested by FQ-PCR and our home-made gene-chip. 52 HEV positive samples tested by FQ-PCR all exhibit positively when tested by our rap- id assay; These results suggested that chip-based detection method was consistent with a reference Fluorescence Quantitative PCR assay.

关 键 词：纳米颗粒 生物素 链霉亲和素 戊肝病毒 基因检测 

分 类 号：Q78[生物学—分子生物学]