Bacterial Imprinting Facilitated By Bacterial Chemotaxis

The development of precise quantitative detection and analysis technology of bacteria has important practical value in food hygiene,environmental health,experimental diagnosis and other fields.But the highly selective separation of target bacteria is the prerequisite for the success of quantitative detection technology of bacteria.As is known to all,the coexistence of advantages and disadvantages of traditional methods cannot meet the increasingly fine requirements of molecular biology for bacterial separation.Therefore,it is of great significance to develop new separation technologies for highly selective separation of target bacteria for quantitative detection of bacteria.As a new technology for isolate bacteria,bacterial imprinting technology is a new method that takes target bacteria as template to synthesize pores with complementary structure with target bacteria through functional monomers and crosslinking agents.The microchannel structure can be specifically bound to target bacteria by matching shape,size and chemical bonding.However,conventional Bacterial imprinting techniques for bacterial separation only treat bacteria as template of Bacterial imprinted polymers(BIPs),while ignoring the biological properties of bacteria as microorganisms,which leads to the undesirable separation effects.The research strategy of this paper is to develop a new method to isolate bacteria by combining bacterial chemotaxis with selectivity of bacterial imprinting polymers.In order to achieve bacterial chemotaxis,the microchannel structures will be introduced into BIPs materials.The microchannel structure will be used for the release of chemotaxis and repellants.In the process of bacterial adsorption,the release of target bacteria chemoattractant and non-target bacteria repellent can promote the selective movement and recognition of target bacteria by avoiding the interference of non-target bacteria and to achieve the selective separation of target bacteria.Objective: Combining the passive recognition of traditional bacterial imprinting technology with the active recognition induced by bacterial chemotaxis,a new method with higher selectivity than traditional bacterial imprinting materials was developed,which can be used for the separation of target bacteria in mixed solution.Method: Bacterial PDMS imprinted membrane was prepared by surface imprinting technology,modified with methyltrichlorosilane,and finally the microchannel PDMS imprinted membrane was obtained by pore-forming on clean PDMS imprinted membrane with microneedle(36 holes).The imprinting side of the microchannel PDMS imprinted membrane was oriented towards the bacterial solution,while the non-imprinted side towards the chemotaxis solution to prepare a device that could release chemotaxis slowly to research the adsorption effect and selectivity of PDMS imprinted membrane in the single bacterial solution and mixed bacterial solution.Result: In this paper,PDMS imprinted membrane which can selectively identify bacteria was prepared by surface imprinting technology.The microchannel PDMS imprinted membrane with chemoattractant can be obtained by microneedle pore-forming method.In a single bacteria solution,target bacteria chemotactic agents can significantly increase the adsorption capacity of target bacteria,while interfering bacteria chemotactic agents can also significantly reduce the adsorption capacity of interfering bacteria.In the fluid of the mixed bacteria,chemotaxis or repellent simultaneously increasing or decreasing interference and target bacteria adsorption cant not improve the selectivity of BIP;Chemotaxis or repellent that can only produce chemotactic or repellent effects on target bacteria or interfering bacteria can increase the selectivity of BIP to target bacteria,thus achieving the selective separation of target bacteria.Conclusion: The method based on bacterial chemotactic agents and microchannel imprinted membrane can realize both passive and active recognition of bacteria,thus improving the selectivity of PDMS imprinted membrane to achieve highly selective separation of bacteria.In this study,a new method was proposed to solve the low selectivity of blotting holes in bacterial imprinting technology,which promoted the development of bacterial imprinting technology in clinical bacterial detection.