Research And Implementation Of POCT Nucleic Acid Diagnostic Integrated Microfluidic Syste

Infectious diseases have always caused serious threats to public health,especially the outbreak of COVID-19 in 2020,due to its strong infectivity and high morbidity,the global pandemic has caused huge losses to the world.Nucleic acid detection methods have been one of the gold standards for infectious disease detection due to their significant advantages such as high detection sensitivity and strong specificity.In particular,POCT(point-of-care test)nucleic acid detection is gaining close attention due to its obvious advantages such as short detection time,high degree of automation and easy operation.Nucleic acid diagnosis mainly includes nucleic acid extraction and nucleic acid amplification detection.In traditional nucleic acid detection,these two parts are often completed independently by two types of instruments,such as nucleic acid extraction equipment and nucleic acid amplification detection equipment.Therefore,the nucleic acid detection steps are cumbersome,the detection efficiency is low,and the dependence on various types of equipment is strong,which is not conducive to the popularization and application of nucleic acid diagnostic methods.Based on the concept of POCT nucleic acid diagnosis and the unique technical advantages of microfluidic chips,research and construct a microfluidic system which can achieve integrated nucleic acid diagnosis and realize the mutual integration of nucleic acid extraction and nucleic acid amplification detection,this is of great significance for improving the overall performance of nucleic acid detection.An integrated POCT nucleic acid diagnostic system that integrates nucleic acid extraction and nucleic acid amplification and detection based on a microfluidic chip has been researched in this paper.First,the solid phase extraction membrane was integrated into the microfluidic chip,and a rapid nucleic acid extraction method based on the solid phase extraction membrane on the chip was studied.Through the matching fluid control,efficient nucleic acid extraction was achieved;Secondly,the nucleic acid amplification microfluidic chip based on RPA isothermal amplification technology was studied and a real-time isothermal amplification microfluidic system was constructed to achieve rapid nucleic acid amplification and detection;Finally,based on the integrated microfluidic chip,the microfluidic chip integrates nucleic acid extraction and RPA isothermal amplification detection.The research has realized a portable POCT nucleic acid diagnostic microfluidic system that integrates nucleic acid extraction,RPA isothermal amplification and real-time detection.The experimental results show that the key performance indicators of the integrated microfluidic system for POCT nucleic acid diagnosis studied in this paper have reached the expected requirements,laying a solid foundation for the rapid diagnosis of POCT nucleic acid diagnosis.