A New Pressure Detection Method Based On Nanosphere Brush Signal Amplification System

Background and ObjectiveDue to the rapid development of life science,in vitro diagnosis methods keep pace with the times,POCT for field testing came into being because of clinical needs,to achieve small,rapid,economic,real-time,simple testing.Real-time detection of(POCT),simply refers to a rapid detection and analysis technology.POCT,as a direction of clinical laboratory discipline,has been developed rapidly.Instant detection technology is a small,fast,economical,and instant Simple test method.Instant detection(POCT),in a nutshell,refers to a rapid detection and analysis technique that has developed rapidly as a direction in clinical laboratory disciplines in the last decade.Due to the low concentration of the target and the complex composition of the biological sample,when the large-scale instrument cannot be used in the field test,how to create a sensitive,specific,simple,and portable detection method is still the main problem that needs to be actively solved in the field of real-time detection.The pressure-based ELISA(PBE for short)developed by Yang’s research group in recent years is based on the current status and needs of in vitro diagnostic techniques.It focuses on two important directions of in vitro diagnostic development—instant detection and accurate diagnosis.,continuous improvement.The airsensing-based biosensing platform developed by Yang’s research group is based on the principle that platinum nanoparticles(PtNPs)have catalase activity and can catalyze the decomposition of H2O2 to produce oxygen,and introduce biotin-derived PtNPs into the immune response.Then,under the action of streptavidin/biotin,it combines with the "sandwich" structure formed in the immune reaction,and generates a gas pressure signal by catalyzing the hydrogenation of the substrate hydrogen peroxide,which is finally determined by a hand-held barometer.The immune signal is converted to a barometric pressure signal.Since the establishment of the air pressure platform,the targets of CRP,PSA,HER-2 protein have been measured successively.The detection results are in good agreement with the ELISA gold standard,and the detection sensitivity is almost the same as that of the ordinary ELISA.And through experiments,it can be seen that the results of non-professionals detecting protein targets through the air pressure platform are not much different from those of the professionals.It can be seen that the air pressure platform meets the requirements of small,simple,fast,and instant detection of immediate detection requirements.Due to the clinical early promotion of early detection,early diagnosis and early treatment,we hope to improve the detection sensitivity of the air pressure platform and enhance the accuracy of the instant detection equipment and the early screening function of the disease.There are many devices for signal amplification,such as carbon nanocage,mesoporous silica,and screening.In this study,a polyacrylic acid ball brush is used as an enzymatic carrier for signal amplification.In combination with a pneumatic platform,a pressure based on a polyacrylic ball brush amplification system is developed.A new method was tested to quantify the H5 NI protein and compared to traditional ELISA results.Secondly,the platinum particles used in the previous pressure platform will be agglomerated and reduced in activity within a certain period of time after synthesis,so the platinum particles used in each test need to be ready for use.This greatly limits the pace of the pressure platform as a true instant detection device for product conversion.Another problem to be solved in this paper is to synthesize a stable and easy to preserve new platinum particle.This study aims to construct a portable,low-cost,simple-to-use,high-sensitivity method for air pressure detection Materials and MethodsAcrylic acid ball brush is an enzyme carrier,which has the function of amplifying the signal.Platinum nanoparticles(PtNPs)has catalase-like activity and can catalyze the decomposition of H2O2 into oxygen,which is the basis of gas production.In this study,a new type of platinum nanoparticles with stable catalytic activity will be redesigned.The acrylic acid ball brush was encapsulated with PtNPs and catalase respectively,and the enzyme amplification effect was realized.the ball brush was coupled with the detected antibody and then introduced into the immune reaction.Then it binds to the antigen-capture antibody structure formed in the immune reaction,catalyzes the substrate hydrogen peroxide to produce gas,produces the air pressure,and finally determines by the hand-held small barometer.The new detection method was used to detect H5 NI protein and compared with the traditional method.The main research methods were enzyme-linked immunosorbent assay(ELISA)and double antibody sandwich ELISA experiment based on acrylic acid ball brush amplification system combined with air pressure platform.ResultsThe new platinum nanoparticles and Catalase were successfully encapsulated into the acrylic acid ball brush,and a new method of air pressure detection based on the amplification system of the acrylic acid ball brush was constructed.In the course of the experiment,the catalytic activity of the new platinum nanoparticles and catalase was monitored,and the method of encapsulating the new platinum nanoparticles and catalase with acrylic acid ball brush was optimized.The experimental results showed that H5N1 protein was measured by pressure detection method based on polyacrylic acid ball brush coated platinum nanoparticles amplification system under the reaction conditions of 200 uL reaction system,reaction time 1 h and 30% H2O2 substrate concentration.The detection sensitivity is 0.2ng/mL.compared with the traditional ELISA method,the current data has not achieved the goal of amplifying the signal.in the later stage,the experimental conditions will continue to be optimized to improve the sensitivity.The pressure detection method based on acrylic acid ball brush wrapped catalase amplification system was used to measure H5N1 protein under the reaction conditions of 200 uL reaction system,reaction time 1 h and 1% H2O2 substrate concentration.The detection sensitivity is 0.01 ng / mL,and the target of amplifying the signal is successfully realized by the current data compared with the traditional ELISA method.Conclusion 1.A novel platinum nanoparticle(PtNPs)was successfully synthesized and the PtNPs was encapsulated into the acrylic acid ball brush and then introduced into the immune reaction system,and a new pressure detection method based on the amplification system of the acrylic acid ball brush was established.The detection sensitivity was 0.2ng/mL.2.The catalase was successfully wrapped into a polyacrylic acid ball brush and then introduced into the immune reaction system to establish a new method for pressure detection based on polyacrylic acid brush amplification system with a detection sensitivity of 0.01 ng/mL.The sensitivity of the conventional ELISA was compared to the target of signal amplification.