Method for fast and sensitive gene detection of pathogenic bacteria by paper-based ambipolar electrode electrochemiluminescence molecular switch system

Abstract

The invention discloses a method for fast and sensitive gene detection of pathogenic bacteria by a paper-based ambipolar electrode electrochemiluminescence molecular switch system. The method comprises the following steps that DNA (deoxyribonucleic acid) molecular fragments of a target virulence gene of the pathogenic bacteria and a PBS solution containing photoswitch molecules and TPrA to obtain a mixed solution; then, the mixed solution is dripped onto pBPE; the front side of the pBPE faces PMT; then, the pBPE is put in a dark box; finally, a pair of driving electrodes of the pBPE are connected to a direct current power supply; driving voltage is exerted; an ECL signal is obtained at an anode of the pBPE; the maximum reproducible luminescence signal observed in 10s is used as an effective signal value; the goal of fast and sensitive gene detection of pathogenic bacteria can be achieved. The method has the advantages that the properties of molecular switches are used; the goal of non-marking detection can be achieved; the sensitivity is high; the cost is low; the building of the detection device is simpler and more convenient.

Description

technical field [0001] The invention belongs to the technical field of biological detection, in particular to a method for rapid and sensitive gene detection of pathogenic bacteria by a paper-based bipolar electrode electrochemical luminescence molecular switch system. Background technique [0002] Genetic testing is a very important method for detecting pathogenic bacteria. Various strategies and techniques, such as real-time quantitative polymerase chain reaction (RT-PCR), nucleic acid sequence-based amplification (NASBA), and loop-mediated isothermal amplification (LAMP), have been developed to detect target pathogens characterization genes and has been used as a standard method in central research laboratories. However, these traditional methods are usually laborious and time-consuming. With the development of biosensing technology, the development of certain optical analysis methods, including colorimetry, fluorescence spectroscopy, chemiluminescence, and electrochemi...

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Problems solved by technology

[0006] However, labeling probes on paper faces several challenges, such as the need for expensive probe labels and the difficulty of removing nonspecifically bound probes, resulting in relatively high background signals

In addition, most paper-based assays are usually based on colorimetric methods, and when the analyte concentration is low, the quantitative or semi-quantitative assays used cannot meet the requirements of high-sensitivity detection

For these reasons, paper-based ECL genetic testing methods are used less

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