Detection Of Lester’s Bacteria Based On IsothermaL Hybridization And Chain Reaction And Enzyme-Linked Immunosorbent Assay

Lester’s bacteria are ubiquitous in our living environment,and Lester’s bacteria exist in most food and food products.Many countries have adopted corresponding control measures and formulated corresponding guidelines to avoid diseases caused by Lester’s infection.Traditional methods of isolation and culture,immunology detection methods are long time,need specific enzyme catalysis,low sensitivity and can only be applied to the laboratory,so we try to establish a new detection method.The detection method established in this experiment does not require temperature change,enzyme catalysis,short time consumption,and does not need professional test personnel operation,so it is suitable for mass detection.Magnetic beads combined with isothermal chain reaction:using the magnetic beads as the detection platform,using the sandwich structure to connect the grasping probe,the target target and the detection probe,the effect of the signal amplification through the HCR reaction,and the visualization of the naked eye detection by the HRP enzyme catalyzed TMB color reaction.The experiment is highly sensitive.The detection of the concentration of desert bacteria is 3 x 10~3CFU/mL,and the concentration of Lester bacteria is 3 x 10~4 CFU/mL.Magnetic beads combined with nanoscale:under the condition of PBS buffer,two valence copper sulfate solution and ConA protein are prepared for nanoscale flowers.Through the binding properties of ConA protein and pathogenic bacteria,the nanoscale flowers are connected with the bacteria.Finally,visual detection is realized by clicking on the test paper strip of chemical reaction point.The established detection method is highly sensitive,and the minimum detection line is 4 CFU/mL.The two detection methods established in this experiment can achieve specific detection of mono Lester bacteria by different detection platforms and different signal output methods,which not only improves the detection limit,simplifies the operation process,low cost,strong anti-interference ability,the detection line can reach pM,and realizes the immobilization of protein.It can be applied to the analysis and detection of Lester bacteria in a variety of samples,which is beneficial to the establishment of a low cost and high sensitive sample detection method,to lay a better foundation for the establishment of new detection methods in the future,and to have a wider foreground in some fields,such as food hygiene inspection,clinical diagnosis and so on.