| Back ground: Cardiovascular diseases still occupied the peak of death rate at present.Acute myocardial infarction(AMI)is the main "killer" and shows a younger trend according to "China Cardiovascular Disease Report 2017".Lacking of effective means for early screening of AMI leads to miss diagnosis as the current basic medical conditions in our country are not perfect.This phenomenon is more serious in rural and remote areas.85% cardiocytes is damaged within 2 hours of AMI,meanwhile mortality risk is elevated if postponing the treatment.Thus,it is extremely important for clinic treatment which diagnosed of AMI early and rapidly.Cardiac troponin I(c Tn I)is used for detecting important marker of AMI as it's specificity of up to 94.78%.Simultaneously,it is used as the clinical marker most commonly.Heart fatty acid-binding protein(H-FABP)is focused widely in recent years as one of the early indicators of AMI with its small molecular weight and can be detected early(within 0-3 h)of acute coronary syndrome.H-FABP is reported to be a marker for ultra-early diagnosis and excluding AMI according to previous studies.Consequently it can make up for the shortcomings of c Tn I which appears later(3-6 h).To detect AMI more accurately and earlier,the c Tn I and H-FABP were tested simultaneously.In addition,inspection methods for AMI should meet real-time detection and the instrument is smart,portable,easy to operate and interpret due to the irregular onset of AMI,.Immunochromatographic assay,named as lateral flow immunoassay(LFIA),is a powerful tool for fast,easy-to-read,low-cost on-site detection of biomolecules.Therefore,the simple,economical and accurate POCT has drawn much attention in recent years.Currently,many labeled probes are used for lateral chromatography,among which colloidal gold is the most commonly used.Colloidal selenium is more stable and nano-enzyme owns higher sensitivity which are different from colloidal gold.However,whether colloidal selenium and nano-enzymes can be used for the detection of AMI,and research about the differences between the prepared kits and colloidal gold are still less.This project aims at developing a kit for detecting AMI rapidly in order to reduce the admission rate of AMI and improve the life quality of patients.Purpose: 1.Preparation and application of lateral flow immunochromatographic strip for H-FABP detection based on colloidal selenium.2.Fabrication and appliance of rapid test kit for c Tn I detection based on nano-enzyme.Methods: 1.Colloidal selenium was prepared by selenite reducing ascorbate with SDS and PEG-20000 as modifiers,its quality control was carried out.2.Preparation lateral flow immunochromatographic strip with colloidal selenium mentioned above.Test the sensitivity,specificity and stability in plasma.Sensitivity of the stips in blood was detected simultaneous ly.And finally,the difference of sensitivity,accuracy and other characteristics between colloidal selenium and colloidal gold test strips under the same conditions was compared.At the same time,its value in clinical was evaluated.3.Magnetic nanoparticle was synthesized by hydrothermal method.It was coupled with c Tn I antibody to obtain antibody-nanozyme complex.Fabrication immunoassay strips for c Tn I detection based on the probe.Afterwards,the sensitivity,specificity and stability in plasma were detected.Ultimately,difference of sensitivity between commercial colloidal gold and nano-enzyme test strips was compared.Results: 1.Colloidal selenium which was clear,spherical and homogeneous was successfully prepared using 0.04 M Vc to reduce 0.32 M H2 Se O3 with 0.25 m M PEG-20000 and 100 m M SDS as the stabilizer.Nanozyme which is fit for the kit was prepared triumphantly with 30 g/L Fe Cl3·6H2O and 75 g/L Na Ac as the substrates.2.Colloidal selenium strip can be used for detect H-FABP.The device exhibited a high sensitivity of 5 ng/m L in plasma.Furthermore,the equipment owned a good specificity and stability.In the meantime,the system showed a high sensitivity of 5 ng/m L in blood.3.Nano-test strips of c Tn I were developed successfully with a sensitivity of 1.5 ng/m L in plasma which meet the clinical need of diagnosis of AMI.The device displied an excellent stability.Moreover,there is no cross reaction with other AMI markers such as H-FABP,CK-MB,and Myo.That is the kit owns outstanding specificity.When compared with commercial strip of colloidal gold,it was found that the sensitivity of nano-test strips possessed about 3.3 times higher than commercial colloidal gold strips.Conclusion: We developed the rapid diagnosis kit of acute myocardial infarction successfully which could be used to detect H-FABP and c Tn I,respectively.The kit owns a high sensitivity,specificity and stability. |
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