Preparation Of Chemiluminescent Hydrogel Bead And Its Application For Acute Myocardial Infarction Biomarkers Detection

Acute myocardial infarction（AMI）,as a common clinical cardiovascular disease,is the main cause of death in the world.Therefore,early accurate diagnosis and timely treatment of AMI is the key to improving the survival rate.Cardiac troponin（c Tn I）is an important component in the diagnosis of myocardial injury Clinical studies have shown that combined detection of c Tn I and cardiac fatty acid binding protein（h-FABP）can improve the accuracy of early diagnosis of AMI.Chemiluminescence immunoassay（CLIA）is a combination of highly sensitive chemiluminescence determination technology and highly specificity immunoreaction,which has been widely applied to the detection of a variety of disease biomarkers,and having the advantages of simple,fast,high sensitive,wide detection range,low cost.Therefore,two kinds of chemiluminescent hydrogel beads were successfully prepared based on chemiluminescence and chemiluminescence energy transfer theory and the prepared hydrogel beads were further used as signal probes to construct chemiluminescence immunoassay to detect AMI biomarkers including c Tn I and h-FABP.（1）Two kinds of chemiluminescence hydrogel beads were prepaed based on chemiluminescence and chemiluminescence energy transfer theory.Firstly,chitosan（CS）hydrogel was prepared,and then gold nanoparticles（Au NPs）,chemiluminescence catalyst and chemiluminescence substrate,were further embedded into the CS hydrogel to form hydrogel complex.Finally,hydrogel complex was treated by strong alkali to form hydrogel beads.The chemiluminescence hydrogel beads can release visible chemiluminescence signals initiated by hydrogen peroxide.CS-Co²⁺-Lu-Au hydrogel bead prepared by using luminol as chemiluminescence substrate can release blue light emmision.CS-Co²⁺-Lu-FL-Au hydrogel bead prepared by using luminol and fluorescein as chemiluminescence substrates can release green light emmision based on chemiluminescence resonance energy transfer theory.The prepared two kinds of hydrogel beads exhibited obviously improved chemiluminescence performance and high stability.（2）Based on the prepared two kinds of chemiluminescence hydrogel beads as signal probes,chemiluminescence immunoassays were constructed for the detection of two important acute myocardial infarction AMI biomarkers,including c Tn I and h-FABP,by using mobile phone as a portable detector.Specific antibodies（Ab）anti-c Tn I and h-FABP can be directly assembled onto the surface of CS-Co²⁺-Lu-Au and CS-Co²⁺-Lu-FL-Au hydrogel beads by using Au NPs as binding sites to obtain CL hydrogel beads(CS-Co²⁺-Lu-Au-Ab and CS-Co²⁺-Lu-FL-Au-Ab)with both good CL activity and immune activity could be obtained.The chemiluminescent immunoassay based on hydrogel beads does not require complex and time-consuming centrifugation or magnetic separation steps,which greatly simplifies the detection procedure.The results show that c Tn I and h-FABP have a good linear relationship at concentration range from 1.0×10^-11to 1.0×10^-5g/m L.Finally,the method was successfully applied to the detection of c Tn I and h-FABP in serum of healthy and AMI patients,which proved its practical application in the diagnosis of AMI.