Visual gene diagnosis of HBV and HCV based on nanoparticle probe amplification and silver staining enhancement.
A visual gene-detecting technique using nanoparticle-supported gene probes is described. With the aid of gold nanoparticle-supported 3'-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 home-made 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.
Wang
Ye-Fu
YF
College of Life Sciences, Wuhan University, Wuhan, Peoples Republic of China.
Pang
Dai-Wen
DW
Zhang
Zhi-Ling
ZL
Zheng
Hu-Zhi
HZ
Cao
Jun-Ping
JP
Shen
Jun-Tao
JT
D015342:DNA Probes; D005784:Gene Amplification; D005814:Genes, Viral; D006046:Gold / Q000737:chemistry; D016174:Hepacivirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D006509:Hepatitis B / Q000175:diagnosis*; D006515:Hepatitis B virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D006526:Hepatitis C / Q000175:diagnosis*; D006801:Humans; D036103:Nanotechnology / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques; D010316:Particle Size; D016622:Silver Staining / Q000379:methods*; D014764:Viral Proteins / Q000235:genetics
D015342:DNA Probes; D014764:Viral Proteins; D006046:Gold