Assay Of DNA And Lysozyme Based On Rolling Circle Amplification And Biological Bar Code Technology

By using of DNA polymerase and nicking endonuclease(NEase),a series of flow injection chemiluminescence(FI-CL)and surface enhanced Raman spectroscopy(SERS)methods were developed including NEase assisted target strand circular process,cascade signal amplification based on rolling circle amplification.These methods were used to detect DNA and lysozyme.Because of the sensitivity,simplicity,rapidity and low-cost,these methods could be easily used for biological sample analysis.The contents of this paper include the following three parts:1.Achieving the detection of target DNA amplification by chemiluminescence strength of the signal,based on the rolling circle amplification(RCA)and flow injection chemiluminescence technology(FI-CL).We determine the concentration of target DNA by means of DNA enzyme catalyzed produced by the luminol-H2O2 system luminous intensity quantitative analysis,by RCA amplification and MBs surface combined with a large number of amplification of DNA enzymes,so as to achieve the purpose of detection of low concentrations of target DNA.This method has a high sensitivity and good selectivity,and it is easy to detect and fast.2.The detection of target DNA is achieved by surface enhanced Raman spectroscopy(SERS),based on nicking endonuclease signal amplification,rolling circle amplification(RCA)and bio bar code signal amplification technology.At the presence of rolling circle amplification,the long-chain DNA can hybridize with fluorescent probes modified bio bar code.The last detection is completed by surface enhanced Raman spectroscopy(SERS).This method has a higher sensitivity selectivity and stability.3.A highly sensitive detection of lysozyme is designed based on circular amplification and bio bar code signal amplification technology.Lysozyme was added to the assembled MB-DNA-bio bar code probe complexes,lysozyme will be got after the cyclic amplification.So the solution of the bio bar code probe is remained,the final surface enhanced Raman spectroscopy(SERS)can achieve a high sensitivity detection of lower concentrations of lysozyme.