| In the struggle between humans and diseases,diagnosis,treatment and prevention are three key links.Among them,the diagnosis technology based on in vitro diagnosis is the most critical,which is the basis of treatment and the basis for formulating prevention strategies.At present,there are many in vitro diagnostic techniques,and conventional techniques include immunodiagnosis,molecular diagnosis,and blood cell diagnostic techniques.Among them,molecμLar diagnostic technology is the technical basis of precision medical treatment,and it is also the fastest sub-industry for in vitro diagnostic growth.In recent years,the gene editing in vitro diagnostic technology based on the development of molecular diagnosis has the characteristics of high speed,good specificity,high sensitivity and easy automation.At present,the technology is still in a rapid development stage,and the micro liquid treatment technology is the core technology in in vitro diagnosis,especially in molecular diagnosis.The research of micro-liquid treatment technology in this subject mainly includes:1.Analyze the existing technology of micro-liquid pipettingBy analyzing the current development status of micro-liquid pipetting technology,the current micro-liquid pipetting technology is researched,including the single-module pipetting pump technology,the application of multi-module integrated pipetting workstation technology and the development of microfluidic technology.Analyze the processing accuracy and key technologies of the micro-liquid pipetting equipment,and obtain the micro-liquid pipetting technology suitable for molecular diagnosis-based on the air displacement liquid pipetting technology.2.Research on air displacement liquid pipetting technology and test platform constructionAccording to the basic principle of air displacement liquid pipetting technology,aiming at the various influencing factors in the micro-liquid pipetting process,the use of liquid level sensing and pipetting real-time monitoring technology,calibration compensation,pipetting speed control and other methods to eliminate or reduce the system Sources of error,etc.Through research and analysis,the mechanical structure of the micropipette pump and the precision control detection technology of the stepping motor are optimized and developed,and a micropipette test platform is constructed.The test platform adopts a three-axis rectangular coordinate structure,which automatically completes the loading of the tips,and compares the liquid level sensing,capacitance sensing,and compensation.Within the pipetting range of 30μL～100μL,the pipetting error is not more than ±3%,and the volume variation The coefficient cv value is not more than 3%;within the pipetting range of 100μL～250μL,the pipetting error is not more than ±2%,and the volume variation coefficient cv value is not more than 2%.3.Development of gene editing detectorComplete the design and construction of the gene editing detector based on the micro-liquid pipetting technology.According to the requirements of the gene editing application,the system is divided into a nucleic acid extraction module,an isothermal amplification module,a fluorescence detection module,and a micro-liquid pipetting module.The quadruple tube is used for extraction.Tube,disposable micro-liquid pipetting cartridge as the detection carrier.The extraction module adopts the uninterrupted pipeline mode,the sample extraction concentration cv is not more than 20%;the temperature control accuracy of the isothermal amplification module is ±1℃;the fluorescence detection module achieves the detection precision between samples and the cv value is not more than 20%,the same sample The detection precision cv value is not more than 5%.4.Performance test and clinical application of gene editing instrument.Through the construction test of gene editing equipment,the biological performance of the equipment was tested through clinical samples.Including nucleic acid extraction and verification testing of sample addition modules to obtain nucleic acid samples with a concentration that meets the requirements.The fluorescence detection module and related sample addition modules were verified by fluorescence detection and passed the control group test.Through the above biological performance test verification,it is preliminarily confirmed that the gene editing instrument is based on the micro-liquid pipetting technology to meet the needs of molecular biology nucleic acid detection,and it can be further tested for new coronavirus,A and B,syncytial virus,etc. |
