Integrated Nucleic Acid Analysis Microfluidic System

Nucleic acid detection technology is increasingly used in life sciences and in vitro diagnostics because of its high sensitivity,specificity,and windowless time.In recent years,the Point-of-Care Testing(POCT)technology platform has been significantly promoted and applied.Due to its obvious advantages such as simple operation,convenience and low cost,POCT nucleic acid analysis is getting more and more attention and attention.The microfluidic chip can continuously and automatically complete the steps of sample preparation,reaction,separation,detection and the like related to the biochemical analysis process by means of a microfluidic network composed of microvalve,micropump,microchannel,and microreactor.Compared with the traditional methods,the microfluidic chip can not only realize the automation and rapid detection process,improve the detection efficiency,but also help to achieve the miniaturization and portability of the device,and is more suitable for POCT on-site rapid detection.The POCT nucleic acid analysis based on the microfluidic chip can significantly improve the comprehensive performance of nucleic acid diagnosis,and has become one of the research focuses in the field of microfluidic chips.In order to achieve efficient nucleic acid analysis,this article studied the rapid extraction of nucleic acids,nucleic acid amplification and detection methods on microfluidic chips.We studied the rapid extraction of nucleic acid microfluidic chip.An integrated nucleic acid extraction chip with reagent storage module was designed.Through the cooperation of the chip and the matching device,the nucleic acid extraction-related sample lysis,nucleic acid binding,washing and washing can be performed sequentially.After several steps,the purified nucleic acid template is finally obtained.We studied the rapid amplification and detection of nucleic acids based on real-time convective PCR(CPCR).In the constant-temperature heating mode,a highly efficient PCR nucleic acid amplification is realized with a self-heating circulation inside the capillary reaction chamber.At the same time,the fluorescence signal in the capillary reaction chamber is tested by a smart phone.The real-time nucleic acid amplification and testing are achieved in 30 minutes.Different types of samples were tested to verify the comprehensive performance of the nucleic acid rapid extraction chip and convection PCR amplification detection system.For blood screening samples,the rapid nucleic acid extraction microfluidic system can simultaneously achieve efficient extraction of HBV,HCV,and HIV nucleic acid templates,and the extraction efficiency is basically equivalent to a commercial semi-automatic nucleic acid extractor.Combined with rapid extraction of microfluidic chips and convection PCR amplification and detection system,a rapid extraction,amplification and detection of multiple gradient concentrations were successfully achieved for H1N1 influenza A virus samples.The detection limit reached the expected requirements.Compared with the traditional nucleic acid analysis system,the combination of the microfluidic nucleic acid extraction chip and the real-time convective PCR nucleic acid amplification detection system can realize efficient and convenient automatic nucleic acid analysis within 50 minutes,and lays a good foundation for POCT nucleic acid diagnosis.