A Visual DNA Microarray for High Sensitivity Detection of Hepatitis B Virus DNA and Single Nucleotide Polymorphisms
Hepatitis B is a major disease which causes serious public health problems worldwide.Herein,a visual DNA microarray was developed for sensitive and specific detection of hepatitis B virus DNA(HBV-DNA) and single nucleotide polymorphisms.5′ end amino-modified oligonucleotide,which was immobilized on microarray surface acted as capture probe,and another part of gold nanoparticle-labeled oligonucleotide were designed to bind specifically to complementary targets HBV-DNAt,and then linked as a single-strand DNA via Taq DNA ligase.The black image of microarray spots,resulting from silver stain on gold nanoparticles,was used to detect visually target HBV-DNA and single nucleotide polymorphisms.Capture probe with single base mismatch was prepared to detect specificity and sensitivity of hybridization on microarray.Target HBV-DNA with different concentration had been tested,the result indicated that this method had good specificity in the discrimination of single nucleotide polymorphisms and showed high sensitivity(about 10 pmol/L).There is also a linear relationship between the signal intensity of microarray spots and the concentration of target DNA at the concentration range of 10-100 pmol/L.For its high sensitivity and good specificity,this visual detecting technique has potential applications in clinical fields.