The Study Of The Double Antibody Sandwich Immuno-PCR Detection Technology Based On Nucleic Acid Beacon Aptamer And Magnetic Agar Microsphere

The detection and quantification of molecules play an essential role in basic discovery research as well as in clinical practice. Technologies that allow specific detection and precise quantification of molecules are evolving to cater to new analyses as well as to improve existing techniques. As a result, novel approaches that challenge traditional methods are being discovered.In this paper, the magnetic microspheres will combine with nucleic acid beacon aptamer as vehicle to bulid a new protein detection technology. For the success of this technology establishment, the experiment was carried out continuously grope and finally established experimental basic flow. This technique is to use magnetic agar microsphere to fix target protein, then combined with nucleic acid beacon aptamers, and complete signal conversion from the target protein to the nucleic acid beacon by elution separation; Using real-time quantitative-PCR technology to take a rapid and quantitative detection of beacon aptamers, also for indirect detection of target protein. In this process, the best dilution multiples and combination time of antigens, antibodies and nucleic acid beacon aptamers, elution separation reagents, methods and sealing effect are optimized, and we carried on the experimental verification of the specificity, repeatability and accuracy of this method. The results showed that all the antigens and antibodies had good positive values. In addition, Their best dilute concentrations are 100. we also measured that the nucleic acid beacon aptamer had good specificity and the best dilution of it was 50, the optimum reaction duration of it was 30 min. Moreover we measured the most appropriate elution reagent of this method was SSC and the best way to elute was alternating elution, et al. Finally it is concluded that this method can be used to recognize the specific target protein, and can reflect the minute amount of target protein, so it is proved to be feasible.This technology has the signal amplification characteristics of nucleic acid beacon aptamer, vector characteristics of magnetic microspheres, and few artificial operation, elution separation as a means of separation, then complete the indirect quantitative detection of target molecules by using real-time quantitative-PCR technology. This technique provides a rapid, sensitive and practical technical method for protein quantitative detection.