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Fluorescent solid-phase biochip detection method based on LED light source

A technology of LED light source and detection method, which is applied in the direction of fluorescence/phosphorescence, measuring device, and material analysis through optical means, which can solve the problems of low intensity, high requirement for fluorescence intensity, and failure to meet the requirements of detection sensitivity. The effect of testing costs

Inactive Publication Date: 2019-04-09
NANJING XIANGZHONG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For solid-phase biochips, if LEDs are used as the light source for detection, the requirements for the fluorescence intensity of fluorescent materials are high. The intensity of commonly used fluorescent materials is too low to meet the detection sensitivity requirements.

Method used

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  • Fluorescent solid-phase biochip detection method based on LED light source

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The preparation method of the nano-silver probe modified by the oligonucleotide chain labeled with the aptamer and the fluorescent material:

[0026] (1) Add 10mmol / L silver nitrate solution dropwise to 30mmol / L sodium borohydride solution under ice bath conditions, the reaction molar ratio of silver nitrate and sodium borohydride is 1:3, keep stirring until the reaction is complete, Then add sodium borohydride solution, the molar weight of the added sodium borohydride is 7% of the molar weight of the above-mentioned sodium borohydride participating in the reaction, continue to stir to room temperature, obtain nano-silver solution;

[0027] (2) Add different aptamer chains and fluorescent material-labeled oligonucleotide chains in a molar ratio of 1:1 to the nano-silver solution obtained in step (1), and let stand for 24 hours, the aptamer chain and fluorescent material The ratio of the total molar weight of the labeled oligonucleotide chain to the nano silver molar wei...

Embodiment 2

[0033] A fluorescent solid-phase biochip detection method based on an LED light source, comprising the following steps:

[0034] (1) Make a biochip: mix the antigen of the test object in PBS buffer solution, apply the sample in the reaction zone of the carrier sheet, and then seal the carrier substrate with calf serum that does not have active groups that are bound to the sample;

[0035] (2) Add mixed standard solutions containing detection targets with different concentration gradients to some of the reaction wells of the above-mentioned biochip, respectively add the sample solution to be tested in the remaining reaction wells, and continue to add detection targets to all reaction wells in sequence The corresponding aptamer prepared in Example 1 and the nano-silver probe modified by the oligonucleotide chain labeled with fluorescent material were reacted at 37° C. for 20 min, washed with washing solution; Mix with 2mmol / L hydroquinone in an equal volume for color development...

Embodiment 3

[0038] Compared with Example 2, in this example, the chromogenic agent in step (2) is replaced by: 10mmol / L chloroauric acid and 10mmol / L tannic acid are mixed in equal volumes for color development, so that the nano-silver probes form nanoparticles polymer. All the other operations are the same as in Example 2.

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Abstract

The invention discloses a fluorescent solid-phase biochip detection method based on an LED light source. The method comprises the following steps of mixing antigens, antibodies, receptors or aptamersof to-be-detected objects in a PBS buffer solution, dispensing the objects to a reaction zone of a carrier substrate, and then sealing active groups not combined with the dispensed objects on the carrier substrate by using calf serum; adding a mixed standard solution containing detected targets with different concentration gradients into part of reaction holes of a biochip, adding a to-be-detectedobject solution into the remaining reaction holes, continuously adding fluorescent material marked monoclonal antibodies or aptamers corresponding to the detected objects into all the reaction holesin sequence for reaction, and carrying out washing with a washing solution; and finally scanning an image by using a fluorescence analyzer of the LED light source and quantitatively calculating the content of the detected objects in the to-be-detected object solution. The detection method can greatly reduce the detection cost, is suitable for popularization and application in the field of base layer detection, and can meet the requirements for simultaneous rapid quantitative detection of multiple indexes in clinical samples, foods and environment samples.

Description

technical field [0001] The invention relates to a biochip detection method, in particular to a low-cost solid-phase biochip detection method for detecting residues of multiple indicators in a sample to be tested by a fluorescence method based on an LED light source. Background technique [0002] Existing biochips refer to microarrays of high-density DNA, antigens, antibodies, cells or tissues coated on immobilized carriers, using the ability of biomacromolecules to specifically recognize each other to arrange them in an orderly manner on a solid carrier On substrates (orifice plates, membranes, glass slides, polymer carriers, etc.), react or hybridize with detection samples and labeled biomolecules at the same time, and a large amount of useful biological information can be obtained through automatic reading equipment. At present, biochip detection methods mainly include fluorescence method, chemiluminescence method, catalytic colorimetric method and so on. Since both chemi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 李周敏李钟卉
Owner NANJING XIANGZHONG BIOTECH