Application Of Nucleic Acid Molecular-coded Probe In Pathogen Detection

Pathogenic bacteria is a kind of pathogenic microorganism,which exists in the environment,food and human body and can cause the outbreak of a variety of diseases.It can cause bacterial infection and lead to many serious complications.Traditional pathogen detection methods have some limitations in sensitivity,specificity,detection speed,cost,and high-throughput detection and analysis of bacteria.Therefore,it is urgent to find new pathogen detection methods with good sensitivity,high specificity,simple operation and high detection flux.For this reason,we successfully developed three detection methods of pathogenic bacteria by using nucleic acid molecular machine-coded nano-probe,and these methods are superior to the traditional detection methods of pathogenic bacteria in terms of detection efficiency and flux.Firstly,we designed a fluorescent probe based on a nucleic acid molecular machine.When the bacteria to be tested are added,They will trigger the movement of the nucleic acid molecule.Under the power of the exonuclease,the fluorescein on the surface of the probe is sheared off.The overall fluorescence intensity of the solution to be tested is enhanced.By measuring the change of the fluorescence intensity of the solution before and after the reaction,single bacteria detection can be achieved ultra-sensitively,and the overall detection time is less than six hours.Second,in order to further improve the efficiency of the nucleic acid molecule machine and reduce the detection cost,we further designed a dual nucleic acid molecular machine detection system.When the pathogen is added,it will trigger the movement of the double nucleic acid molecule machine,and.It can sensitively detect the pathogens in the solution by comparing the output signal changes based on colorimetry.Since probes have no fluorescence modification and double nucleic acid molecular machine are used,the whole detection cost is greatly reduced and the detection time is further shortened,and the whole detection process can be completed within four hours.Finally,we constructed a droplet coding technology based on nucleic acid molecular machine to achieve ultra-high-throughput detection of pathogens.The fluorescent signal is encoded by a nucleic acid molecule machine that responds to bacteria specifically.Combining the fluorescently encoded probe with the droplet based microfluidic system and flow cytometry greatly increases the detection flux of pathogens.