A Study On SPR-Based Gene Chip For Rapid Pathogen Detection

Objective: we constructs a new gene chip detection system with SPR(Surface Plasmon Resonance ) technology. SPR biosensor is fast, sensitive, effective, and without being labeled or purified .We wish to take the advantage of these characteristics to make our gene chip detection system be simple in equipment, easy in technique, high in sensitivity and wide in application and so on . With these advantages, the new detection system of SPR-based gene chip could be easily used in clinical practice, common laboratories or even complicated situation .Method: 1. In accordance with the literature report and the clinical investigation, characteristic nucleic acid marker of some bacteria, virus gene were determined, and the amplification primers were designed . The amplification methods of these pathogens were established ; 2. The specific oligonucleotide probes of target pathogens were designed and screened by bioinformatical technology using computer. Specificity and reliability of the probes were verified by ECL method ; 3. 50nm two-dimensional nanogold film was plated on general glass slide. We have found two methods for plating nanogold film , and compared their results ; 4. Oligonucleotide probe was fixed by SAM(Self-assembled Monolayer) technology , and the fixing time , probe concentration was optimized ; 5. The nanogold film chip fixed the detection probe, positive control and negative control was used to identify clinical samples; 6. The detection performance of this chip was tested by SPR system; 7. Clinical samples were processed by Chelex-100 , and detected by SPR system . The results...