Homogeneous analysis method for visually detecting multiple target objects based on click reaction signal amplification and corollary equipment thereof

Abstract

The invention discloses a homogeneous analysis method for visually detecting multiple targets based on click reaction signal amplification and matched equipment thereof. The method comprises a homogeneous reaction part, a visual imaging part and a machine visual recognition part, the homogeneous reaction part selects different amplification schemes according to different requirements of a to-be-detected object on sensitivity, a click reaction multi-step signal is amplified for immune reaction, and matched instruments and equipment are used for detecting the sensitivity of the to-be-detected object. The micro-fluidic chip and machine vision software are organically combined, the micro-fluidic chip is introduced, the reaction system is reduced, and compared with the previous reaction in a centrifugal tube, the sensitivity and accuracy of the method are improved by the click reaction form and the micro-fluidic chip.

Description

technical field [0001] The invention belongs to the fields of food safety, in vitro diagnosis and environmental monitoring, and in particular relates to a homogeneous analysis method and supporting equipment for multi-target detection based on click reaction signal amplification and visualization. Background technique [0002] With the rapid development of science and technology and the improvement of people's living standards, people have higher and higher requirements for health-related factors such as medical care, food, and the environment. Rapid detection has the advantages of simple operation, low cost, and suitable for on-site analysis. In vitro diagnosis, Food safety, environmental monitoring and other fields have been widely used. Therefore, the development of rapid detection methods and supporting equipment in related fields has important practical significance for protecting people's lives and health. Most of the traditional detection methods rely on large-scale ...

AI Technical Summary

Problems solved by technology

For example, the inventor previously applied for a multi-channel particle detection device and detection method (CN111413264A) for detecting micron particles. Combined to achieve accurate detection of a single target with high sensitivity, but the detection process is cumbersome and requires magnetic separation and repeated washing, which makes it impossible to achieve no-washing and one-step detection

For another example, the inventor previously applied for a biosensing detection method based on the change of the resistance of the microchannel caused by the change of the insulating microsphere concentration (CN112595759A). This invention provides a simple structure and low cost supporting device. A constant voltage / current is applied to both ends of the microchannel, thereby generating electroosmotic flow, changing the state of the microspheres in the channel, resulting in a change in the resistance value, and then indirectly obtaining the content of the target to be measured, but the detection process needs to be washed repeatedly, and it is impossible to achieve multiple targets at the same time detection, which reduces the detection efficiency and increases the detection cost

The applicant also constructed a resistance biosensing detection method (CN 112415058A) based on the state change of insulating microspheres in the early stage. This method does not require washing and is a homogeneous reaction mode, but this method cannot achieve multiple targets. Simultaneous detection of

At the same time, the previous applicant applied for an analysis method based on magnetic separation to detect multiple targets at the same time (CN112964868A). Polymer insulating microspheres are used as signal probes, combined with magnetic separation immunoassay method, to realize simultaneous detection of multiple targets, but this method requires multi-step magnetic separation, which cannot realize one-step detection without washing, and cannot react with clicks Combining to achieve synchronous amplification of multiple detection signals

Moreover, the stability of this method needs to be further improved. The reason is that the biochemical reactions involved in the previous patent application are carried out in centrifuge tubes, the final reaction volume is relatively large, and the volume required for microscopic imaging is small. Taking a small part of the large-volume sample for counting will lose the signal on the one hand, and on the other hand, taking a small amount of sample from the large-volume sample for detection will easily lead to a decrease in the accuracy of microsphere counting, thereby affecting the entire method stability

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