The Establishment Of Direct Direct UDG-LAMP-GoldMag Lateral Flow Immunoassay For A Nucleic Acid Detection

With the improvement of people’s awareness of disease prevention,the in vitro diagnostic barrier has become a trend,that is,early diagnosis of disease tends to be more and more community-based management and even home self-examination is promoted.Therefore,there is an urgent need for a fast and convenient disease detection suitable technology,which has a certain significance for the development of a prevention-oriented "healthy China".In this study,a direct amplification rapid nucleic acid detection method,namely "Direct UDG-LAMP-GoldMag Lateral Flow Immunoassay " was established based on the gold magnetic particle chromatography system combined with loop-mediated isothermal amplification(LAMP).This technology can simultaneously realize the detection of genetic polymorphism SNP and pathogenic microorganism nucleic acid,and an anti-pollution system is introduced into this technology.The detection technology first takes the SNP site of the MTHFR C677 T gene polymorphism as the detection target,establishes a multi-sample type direct expansion detection technology for whole blood and buccal swabs,and then through system design,it overcomes the loop-mediated isothermal and high-efficiency amplification but is easy to contaminate The UDG anti-pollution system is introduced,and finally the stability and effectiveness of the detection technology are analyzed through the performance evaluation of the system.The detection can be completed in about 25 minutes,and the sensitivity can reach 0.16ng/μL.Based on this detection technology,this thesis establishes a rapid detection method for pathogenic microorganism nucleic acid based on the conservative region ORF locus and N protein gene locus of the new coronavirus.This method can complete virus detection in about 25 minutes,and the system is optimized.The sensitivity is verified by using fake viruses and national standard products,and the detection sensitivity can reach 100 copies?m L.At the same time,different types of coronaviruses(MERS,SARS)are used as templates for specific performance evaluation,and the results show that the detection method has good specificity.Finally,the paper analyzes the feasibility of "Direct UDG-LAMP-GoldMag Lateral Flow Immunoassay " nucleic acid detection technology through the detection of clinical samples of MTHFR C677 T gene.In the paper,300 clinical samples were collected,including 200 whole blood and 100 oral swabs,and sequencing was used as the gold standard for comparative analysis.The consistency rate of the test results was 100%,indicating that the test technology has good applicability and reliability.This detection technology can realize nucleic acid extraction-free,high sensitivity,short time-consuming,simple and convenient operation,and provides a new choice for medical care point detection and even future home self-inspection.