Chemiluminescence Immunoassay On MAGICL6000 For Rapid Detection Of Cardiac Markkers

Chemiluminescence immunoassay(CLIA)is an in vitro diagnostic(IVD)method that combines immune reaction with chemiluminescence analysis technology.Its combination with automatic chemiluminescence analyzer has significant advantages such as high specificity,sensitive detection,wide application range,high precision and large throughput.In recent years,the rapid development of domestic brands,constantly seize the market to shake the market monopoly of foreign products in this field.MAGICL6000 is a domestic self-developed automatic chemiluminescence analyzer,which designed for emergency department and clinical departments.Due to its automatic capping blending and hematocrit detection module,it can accurately detect analytes in whole blood samples.This gives it a great advantage in the application of acute myocardial infarction(AMI),the most urgent clinical condition in the emergency department.As one of the most important cardiac markers in the diagnosis of myocardial infarction,troponin I(c Tn I)has the characteristics of high specificity and high predictive value,which can help doctors reduce the blind spot in diagnosis,make diagnosis earlier,and optimize clinical decision making and prognosis assessment.Therefore,based on MAGICL6000 chemiluminescence immunoassay platform,the development of domestic high-quality cardiac marker detection kit represented by c Tn I is not only of vital significance for the accurate diagnosis of clinical myocardial infarction,but also can significantly reduce the detection cost and produce significant social and economic effects.The main work of this paper is as follows:1.Preparation of main raw materials: a.Screening strategy of antibodies.By querying c Tn I fragment information,different protein fragments were prepared and all protein fragment antibodies were screened out,with complete site coverage and without missed detection.b.Preparation and screening of magnetic beads,selecting stationary phase carriers suitable for rapid detection by preparing magnetic particles with different particle sizes and load.c.Studies on the structure of acridine ester mainly focus on: The improvement of acridine ester luminescence efficiency can improve the sensitivity;the improvement of solubility of acridine ester could improve labeling efficiency and reduce non-characteristic adsorption;the improvement of hydrolysis stability can enhance the stability of the reagent.2.Research on coating and signal amplification: a.The magnetic beads with different particle sizes were prepared,and the coating parameters and feeding amount were optimized.b.Study on the labeling amplification strategy of acridine ester,and investigate the signal amplification effect of different nanoparticles,including polystyrene microspheres,metal nanoparticles and gel microspheres.3.Establishment of protocol for whole blood samples analysis: a.Non-hemolysis protocol: Red blood cell antibody is used to separate red blood cells and eliminate the interference of red blood cells.b.Hemolysis protocol: Different hemolysis agents and whole blood ratio were selected to optimize the rapid lysis of red blood cells,and the interference of hemoglobin was eliminated through time-resolved luminescence curve.c.Optimization of calibration parameters of whole red blood cell hematocrit to achieve accurate detection of chemiluminescence method.4.Precision research: a.Modular research for repeatability improvement,including single factor analysis of sample adding module,blending module,incubating module,cleaning module and measuring module.Locating to the cleaning station module was the key factor of repeatability.The random error of the module was controlled and improved by establishing internal standard method.b.Study on system precision,the operation of the system at different temperatures was investigated,and the system precision was improved by improving the temperature control system and heat dissipation system.5.Performance evaluation of the established method: a.Sensitivity study;b.Study of linear range and hook effect;c.Study of interference substances;d.Study on accuracy methodology comparison;e.Studies of consistency for different sample types.In summary,we have developed a matching myocardial marker reagent based on the Getein(?) automatic chemiluminescence instrument MAGICL6000.Through the preparation of main raw materials and optimization and innovation of technological reaction parameters,the reagent has the advantages of short detection time,good repeatability and high accuracy.In addition,the in-depth study of the automatic chemiluminescence system showed that the instrument and reagent have good coordination and system precision,which performance can meet the requirements of clinical application.