Development And Research Of Integrated Fully Enclosed Microfluidic Biological Reagent Reaction Device

Nucleic acid amplification detection is a major detection method for infectious disease pathogens at this stage,but at present,nucleic acid testing needs to be carried out in a professional laboratory,time-consuming and laborious,so in order to improve the ability to deal with sudden epidemics and make up for the shortcomings of traditional testing,this project has developed a timely testing equipment that integrates biological sample pretreatment,nucleic acid amplification and testing,which is of great significance to alleviate the pressure of epidemic prevention.The main work of this article is as follows:(1)Different sample pretreatment and nucleic acid amplification methods were compared and analyzed,and a biological reaction chip was designed,which used chemical cleavage method and magnetic bead method as the nucleic acid sample pretreatment scheme,and fluorescence quantitative PCR method as the pathogen amplification detection scheme.The chip can achieve the function of integrating sample pretreatment,nucleic acid amplification and detection.(2)Analyze the temperature transfer process of the amplification reaction.The thermal resistance of the reactor is calculated,the contact thermal resistance heat transfer model is established,and the finite element analysis software is used for simulation analysis.From the three aspects of contact area gap ratio,contact surface roughness,and contact surface filler,the influencing factors of contact thermal resistance are explored,and a method to reduce contact thermal resistance is proposed,and the finite element simulation software is used for simulation analysis,and finally the temperature difference between the two sides of the contact interface is controlled within 0.1°C.(3)For the polymerase chain reaction nucleic acid amplification temperature requirements,the PID control scheme,fuzzy PID control scheme,radial basis(RBF)neural network control scheme and fuzzy RBF neural network PID control scheme are designed,and the simulation comparison analysis is carried out by simulink,the fuzzy RBF neural network PID control scheme has the corresponding speed,and the set temperature value can be reached within 14 seconds,and there is no overshoot,so the bioreaction amplification system temperature cycle control scheme is used.(4)Build an integrated biological reaction equipment,and use the influenza B commercial nucleic acid detection kit for the whole machine test,the test shows that the integrated biological reaction equipment can reproduce the nucleic acid amplification S-type curve,which shows that the temperature control module and fluorescence detection module of the equipment meet the design requirements.The samples were diluted with different concentration gradients and repeated tests were repeated to obtain equipment to detect sample concentrations at a minimum of 10~2copies/μl.In summary,this paper has developed a nucleic acid detection equipment that integrates sample pretreatment,amplification and detection in response to the current needs of pathogen detection.The device can detect a variety of pathogens at the same time,simplifying the detection steps,shortening the detection time,and providing a new analytical method for inhibiting the spread of infectious diseases and diagnosing infectious diseases.