Construction And Application Research Of A Microfluidic Chip-based Automated Multiplex Assay Platform

In the routine clinical diagnosis of chronic and common diseases,multiple biomarkers associated with the disease are usually tested.At present,the traditional multiplex assays mainly include lateral flow immunochromatographic test strip technology,liquid phase microarray technology,and real-time fluorescence quantitative PCR technology.However,the above methods usually suffer from low detection efficiency,expensive equipment cost,large size,difficulty in coding,and high reagent consumption,which to a certain extent limit the application of multiplex assays.Therefore,the development of portable and rapid multiplex assays with high sensitivity and specificity is an inevitable trend for the future development of in vitro diagnostics.This thesis addresses the problems of traditional multiplexed assays and builds an automated multiplexed assay platform based on microfluidic chip technology combined with embedded technology,molecular hybridization technology and microelectromechanical processing(MEMS)technology to achieve simple,portable,sensitive and specific multiplexed assays.The details are as follows:Firstly,the research background and significance of multiplexed detection are systematically introduced,the current status and problems of traditional multiplexed detection research are analyzed and explored,and the problems of multiplexed detection are identified through its development status,which leads to the main research content of this thesis.Secondly,the development background and applications of microfluidic chips,as well as common preparation methods are comprehensively introduced.The microfluidic chip for multiplexed assays was designed and fabricated,and through its functional validation and optimization,high accuracy capture of targeting microspheres of various sizes was achieved,which provided a good experimental basis for subsequent multiplexed assays.Finally,a multiplex assay protocol was designed on the basis of the multiplex chip,and an automated multiplex assay platform was built for the qualitative and semi-quantitative detection of human immunodeficiency virus(HIV),hepatitis B virus(HBV)and hepatitis C virus(HCV).In this thesis,a multiplex assay chip was designed to realize the precise separation of different size targeting microspheres by using the principle of size separation combined with an automated experimental platform,thus realizing the simultaneous detection of multiple indicators,which has certain clinical application value.