A rapid and specific bacterial detection method based on cell-imprinted microplates

Abstract

Bacterial detection has attracted substantial interest in recent years owing to its importance in biology, medical care, drug discovery, and public health. For such applications, bacterial cell-imprinting technologies are regarded as potential methods, as they can fabricate artificial tailor-made receptors for cellular recognition. In comparison to conventional methods, which generally require a few days for bacterial determination, cell-imprinted polymers can save a substantial amount of time. Here, we report a high-throughput bacterial detection method based on a cell-imprinted 96-well microplate. The fabrication of the bacterial cell-imprinted polypyrrole and nafion complex was accomplished on a gold nanoparticle-coated microplate. The cell-imprinted polymer complex on the microplate can spontaneously rebind and specifically detect target cells with high selectivity in a short time frame (within 30 min). Furthermore, the microplates could discriminate particular target Escherichia coli O157:H7 cells from bacterial mixtures. This simple method may be used for a variety of applications such as clinical testing, food safety, and continuous environmental monitoring.

Evaluation of Surface Structure of Escherichia coli Using Polypyrrole Matrix

“…Cell-imprinted microspheres were obtained through lysis and overoxidation reaction of PPy by immersing in 0.1 M NaOH aqueous solution. 16,17 The overoxidation of PPy led to the removal of anionic species due to the loss of electrostatic interactions and curing the film. 2 As a result, the complementary cavities of E. coli cells' surface were successfully formed on the overoxidized PPy film by the cell imprinting technique.…”

“…Gold-coated microbeads, the surface of which was modified with a self-assembled aminothiophenol monolayer, were dispersed in 20 mL 25 vol% aqueous ethanol containing 2.0 μL nafion. 16,17 After ultrasonication for 20 min, the dispersion was immediately mixed with 30 μL pyrrole and 5.0 mL of E. coli O157:H7 suspension (3.8 × 10 9 cells mL -1 ), and stirred for 40 min. Subsequently, polymerization was induced by oxidation with (NH4)2S2O8 for 12 h at room temperature.…”

Imprinted Polymers as Synthetic Receptors in Sensors for Food Safety

“…Imprinted polymers have been extensively researched for the extraction of toxins, which has also led to the application of these synthetic receptors in sensors [174][175][176][177]. Furthermore, bacteria detection using IPs in sensing devices has also been exploited [28,30,[178][179][180]. The use of synthetic receptors as recognition element in food safety sensing exhibits the potential of becoming a fast, sensitive and cost-effective technology in contrast to the traditional analytical methods [181][182][183].…”

Conjugated Oligo- and Polymers for Bacterial Sensing

“…The specificity of the imprinted material likely arises from multiple interactions with chemical groups in the cavity as well as from shape and size complementarity to the imprinted bacterial cell. The same basic technique was later used to fabricate cell-imprinted microplates used as biorecognition elements in an ELISA detection assay (Shan et al, 2018). The assay showed high selectivity and sensitivity for the imprinted bacterial cells, demonstrating that CPs as cell-imprinted matrices can complement or even replace biological receptors as superior recognition elements in biosensor applications.…”