摘要
建立一种基于纳米金生物条形码技术的二噁英快速筛检方法.利用二噁英诱导激活的芳香烃受体复合物,特异识别以纳米金为报告基团的二噁英反应探针,该探针被释放后进行条形码放大,通过纳米金银染技术增强识别信号,并记录其吸光度值,从而可以简单灵敏地快速筛检二噁英类化合物.在一定的反应时间和浓度范围内(10-14～10-10mol/L),溶液的吸光度值与2,3,7,8-四氯二苯并二噁英(2,3,7,8-tetrachlorodibenzo-p-dioxin,TCDD)浓度之间呈正相关,方法检测限为0.01pmol/L,变异系数为5%～8%.用纳米金生物条形码(Nano-Barcode,nanoparticle-based bio-barcode)方法和现有的生物分析方法(CALUX,chemical-activated luciferase gene expression)分别测定TCDD标准品,并绘制剂量效应曲线.结果表明,本方法灵敏度高,线性范围宽,重复性好.本研究纳米金生物条形码方法的检测灵敏度高于CALUX将近5倍(EC50分别为4×10-12mol/L和2×10-11mol/L),检测限较CALUX降低了10倍(检测限分别为1×10-14mol/L和1×10-13mol/L),变异系数分别为5%～8%和15%～30%.
We have established a nannparticle-based bio-barcode method for the prescreening of dioxins. The formation of AhR-Amt complex triggered by dioxin resulted in a high-affinity to DRE. The DRE probes were completely released as bio-barcodes and amplified by nanoparticle based silver enhancement, then the absorbance of TCDD（2,3,7,8- tetrachlorodibenzo-p-dioxin） were recorded as quantitative signals. The dose- dependency of the assay range between 10^-14mol/L to 10^-10mol/L for soluable products. The TCDD detection limit was 0.01 pmol/L with a coefficient of variation as 5 %-8 % . Compared with CALUX（ chemical-activated luciferase gene expression） bioassay, the sensitivity of our Nano-Barcode method was approximately 5 times higher with ECso of 4×10^-12 mol/L, and the detection limit was 10 times lower as 1×10^-14 mol/L. In addition, our method exhibited a better reproducibility of CV as 5 %-8 % compared to 15 %-30 % of CALUX.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2009年第2期188-192,共5页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金资助项目(No.20377017)~~
关键词
纳米金颗粒
银染增强
二噁英类化合物
四氯二苯并二噁英
生物条形码
多环芳烃受体
毒性当量
gold nanoparticle
silver enhancement
dioxin-like chemicals
2,3,7,8- tetrachlorodibenzo-p- dioxin （TCDD）
bio-barcode
aryl hydrocarbon receptor（AhR）
toxic equivalency （TEQ）
