Nanogold-based Gene Chip for Rapid Pathogen Detection / 中华医院感染学杂志

ABSTRACT

OBJECTIVE A practical gene chip which aimed to detect and identify pathogens rapidly and exactly is developed on the basis of patent technology of nano-enlargement-detection. METHODS Oligonucleotide probes for the specific gene fragments of target pathogens were designed and immobilized on gene chip.Target sequences were labeled by nanogold as reporter materials.After hybridization,its results were recorded by the interaction between nanogold and silver which amplified the hybridization signal to form brown particles,which could be detected by naked eyes. RESULTS The probes designed were all of strong specificity and great reliability possessing identity of hybridization conditions.The reaction time for marking could be decreased by properly raising the ratio of nanogold and nucleic acid and the speed of labeling reaction could be fastened significantly by gentle agitation.A better hybridization results could be obtained when the samples were hybridized for 8 hours at 45℃ with 0.8 mol/L ionic strength,and then strictly rinsed.Furthermore,the hybridization efficiency could be increased remarkably by slight circumgyratation.A better chromatic effect resulted from the reaction way in 3min?3 at 37℃.The sensitivity of gene chip assays in this test could reach to 100 fmol/L.Compared with traditional detection approach,detection by the chip displayed such advantages as speediness and simplicity and the detection results could be easily recognized by naked eyes. CONCLUSIONS The chip detection technology has met the demand of design exhibiting high sensitivity,strong specificity,and easy operation without special device and showing a promising prospect.