Visual gene diagnosis of HBV and HCV based on nanoparticle probe amplification and silver staining enhancement
Visual gene diagnosis of HBV and HCV based on nanoparticle probe amplification and silver staining enhancement
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DOI：
10.1002/jmv.10379
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发表时间：
2003-06-01
影响因子：
12.7
通讯作者：
Shen, JT
中科院分区：
文献类型：
--
作者：
Wang, YF;Pang, DW;Shen, JT
描述了使用纳米颗粒支持的基因探针的视觉基因检测技术。以金纳米粒子负载的T端巯基衍生寡核苷酸作为检测探针，固定在玻璃表面的5'端氨基衍生寡核苷酸作为捕获探针，通过基于高灵敏度“纳米扩增”和银染色的夹心杂交，对目标DNA进行目视检测。采用自制的HBV、HCV和HBV/HCV基因芯片，采用金/银纳米颗粒染色放大法检测感染者血清样本中不同基因型的乙型和丙型肝炎病毒。目前的可视化基因检测技术因其灵敏度高、特异性好、简单、快速和廉价而可以避免已报道方法的局限性。这项技术在很多领域都有潜在的应用前景，特别是多基因检测与基因芯片的检测相结合将会在临床上得到应用。此外，首次使用共振瑞利光散射（RLS）光谱来判断和监测基因探针在金纳米颗粒表面的固定情况。
A visual gene-detecting technique using nanoparticle-supported gene probes is described. With the aid of gold nanoparticle-supported T-end-mercapto-derivatized oligonucleotide serving as detection probe, and 5'-end-amino-derivatized oligonucleotide immobilized on glass surface acting as capturing probe, target DNA was detected visually by sandwich hybridization based on highly sensitive "nano-amplification" and silver staining. Different genotypes of Hepatitis B and C viruses in the serum samples from infected patients were detected using homemade HBV, HCV, and HBV/HCV gene chips by the gold/silver nanoparticle staining amplification method. The present visual gene-detecting technique may avoid limitations with the reported methods, for its high sensitivity, good specificity, simplicity, speed, and cheapness. This technique has potential applications in many fields, especially in multi-gene detection gene chips coupled with the detection will find applications in clinic. Additionally, resonance Rayleigh light scattering (RLS) spectroscopy is used, for the first time, to judge and monitor the immobilization of gene probes on gold nanoparticle surfaces.