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Homogeneous analysis method based on micro-channel resistance change caused by state change of insulating microspheres

A microchannel and phase analysis technology, applied in the detection field, can solve the problems of complex principle, high cost and poor stability of the homogeneous determination method, and achieve the effect of stable and reliable method, good accuracy and strong anti-interference ability

Active Publication Date: 2021-04-02
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to solve the problems of the existing homogeneous measurement method, such as complex principle, high cost, and poor stability, and provide a low-cost, good stability, and easy-to-operate method that changes the resistance of the microchannel based on the state change of insulating microspheres. Homogeneous analysis method to realize rapid on-site detection of a series of targets

Method used

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  • Homogeneous analysis method based on micro-channel resistance change caused by state change of insulating microspheres
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  • Homogeneous analysis method based on micro-channel resistance change caused by state change of insulating microspheres

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Embodiment 1

[0050] Example 1 Construction of a homogeneous analysis device and a microfluidic chip

[0051] Such as figure 1 As shown, a homogeneous analysis device based on the change of the dispersed state of the insulating microspheres causes the change of the resistance of the microchannel, the device includes a first conductive cell and a second conductive cell, and the first conductive cell and the second conductive cell are arranged between There are microchannels, the inner diameter of the microchannel is 10-500 μm, and the length is 0.5-10 mm. The inside of the first conductive cell and the second conductive cell are respectively provided with a positive electrode and a negative electrode, and the positive electrode and the negative electrode are Connect the positive and negative poles of the DC power supply through wires respectively to form a closed-loop circuit. The closed-loop circuit is equipped with a detection instrument and a power output controller. The detection instrum...

Embodiment 2

[0053] Example 2 Correlation between microchannel resistance and dispersion state of insulating microspheres

[0054] To explore the correlation between the resistance of the microchannel and the dispersion state of the insulating particles, the biotinylated PS microspheres were mixed with different concentrations of streptavidin, and added to the conductive cell 1, PBS was added to the conductive cell 2, and the voltage was applied. , while recording the current in the loop, as follows:

[0055] Test 1) The inner diameter of the microchannel is 25 μm, and the length is 1 mm; the diameter of biotinylated PS microspheres is 2 μm, and the concentration is 100 μg / mL; the concentrations of streptavidin are 0, 10, 20, 30, 40, 50, 60 μg / mL; mix equal volumes respectively, incubate for 15 minutes, take 300 μL of the reaction solution and add it to the conductive cell 1, and add 300 μL PBS solution to the conductive cell 2; apply a 60v DC voltage to measure the current in the circuit....

Embodiment 3

[0067] Embodiment 3 adopts homogeneous analysis method to detect chloramphenicol

[0068] The detection principle of chloramphenicol is as follows: in the conductive cell 1, PS microspheres modified with chloramphenicol antibody, PS microspheres modified with chloramphenicol-BSA conjugate, and chloramphenicol in the sample undergo a competitive immune reaction. When there is no chloramphenicol in the sample, the PS microspheres modified by the chloramphenicol antibody and the PS microspheres modified by the chloramphenicol-BSA conjugate combine to form a complex, thereby increasing the proportion of particles in the aggregated state in the solution; when the sample When there is chloramphenicol in the medium, the PS microspheres modified by chloramphenicol and chloramphenicol-BSA conjugates will compete to bind the PS microspheres modified by chloramphenicol antibody, resulting in a decrease in the proportion of particles in the aggregated state. The higher the content, the le...

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Abstract

The invention discloses a homogeneous analysis method based on micro-channel resistance change caused by state change of insulating microspheres, which comprises the steps of respectively modifying biological recognition molecules on the surfaces of the insulating microspheres, adding a target object to be detected for biological reaction, and applying constant voltage or current to the two ends of the micro-channel to form a closed-loop circuit, wherein the mixed solution flows through the micro-channel under the driving of electroosmotic flow, the resistance of the closed-loop circuit is increased when the insulating microspheres pass through the micro-channel, the rising amplitude is related to the aggregation state of the insulating microspheres, and the content of the target object tobe detected can be indirectly obtained by detecting the change value of the current or voltage of the closed-loop circuit. The method does not need any washing and separation steps, only relates to one-step biochemical reaction, greatly shortens the analysis time on the premise of ensuring the accuracy, has the advantages of accuracy, rapidness, low cost and the like, and has a good application prospect in the fields of food safety, in-vitro diagnosis, environmental monitoring and the like needing on-site rapid detection.

Description

technical field [0001] The invention belongs to the detection field, and relates to a homogeneous analysis method, in particular to a homogeneous analysis method based on the change of microchannel resistance caused by the dispersion state change of insulating microspheres. Background technique [0002] The development of accurate, easy-to-operate, fast and low-cost detection methods and supporting equipment is of great significance in the fields of food safety, in vitro diagnostics, and environmental monitoring. An efficient strategy utilizes a format that does not require separation of the specific recognition component and its complex with the target, ie a homogeneous assay format. Such methods rely on the principle that a detection signal is turned on or off depending on a specific binding reaction. In contrast, heterogeneous assay formats rely on physical separation of bound and free specific recognition components, involving extensive washing steps that are time-consu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/12G01N27/416G01N33/53G01N33/543G01N33/553B01L3/00
CPCG01N27/128G01N27/416G01N33/5304G01N33/54313G01N33/54326G01N33/553B01L3/502707B01L2200/10
Inventor 陈翊平何慧禹聂荣彬王知龙
Owner HUAZHONG AGRI UNIV
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