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Instrument-free enzyme-linked immunosorbent assay method

A technology of enzyme-linked immunosorbent assay and determination method, which is applied in the field of instrument-free enzyme-linked immunosorbent assay, can solve the problems of inability to use on-site analysis and instant detection, high analysis cost, and high price, so as to reduce analysis cost and broad application prospects , The effect of improving the detection sensitivity

Inactive Publication Date: 2019-07-23
GUILIN UNIVERSITY OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these optical instruments are expensive and bulky, resulting in high analysis costs of traditional ELISA methods, and cannot be used for on-site analysis and instant detection (Point-of-Care Testing, POCT)

Method used

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  • Instrument-free enzyme-linked immunosorbent assay method
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  • Instrument-free enzyme-linked immunosorbent assay method

Examples

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Effect test

Embodiment 1

[0022] Use the instrument-free enzyme-linked immunosorbent assay method of the present invention to detect 1 pg / mL HOP antigen and blank samples (blank, buffer solution without analyte). Such as figure 1 Shown, the specific steps of this embodiment are:

[0023] Step 1: Add 100 μL of 0.2 mg / mL HOP antibody (prepared in 12 mM sodium bicarbonate-sodium carbonate buffer solution, pH 9.5) to one well on a 96-well plate, and let stand overnight at 4°C in the refrigerator. Discard the liquid and shake dry. Fill each well with washing solution (10 mM sodium dihydrogen phosphate-dipotassium hydrogen phosphate buffer solution, pH 7.2), let it stand for 40 s, and then discard it. After repeating this 5 times, put the well upside down on the flattened roll paper Pat dry.

[0024] Step 2: Add 100 μL of 15 mg / mL bovine serum albumin (BSA) (prepared in 12 mM sodium bicarbonate-sodium carbonate buffer solution, pH 9.5) to a single well and let stand at room temperature for 5 h. Discard t...

Embodiment 2

[0031] The HOP antigen with a concentration range of 0.1 fg / mL to 100 pg / mL is detected by using the instrument-free ELISA method of the present invention. Such as figure 1 Shown, the specific steps of this embodiment are:

[0032]Step 1: Add 100 μL of 0.2 mg / mL HOP antibody (prepared in 12 mM sodium bicarbonate-sodium carbonate buffer solution, pH 9.5) to one well on a 96-well plate, and let stand overnight at 4°C in the refrigerator. Discard the liquid and shake dry. Fill each well with washing solution (10 mM sodium dihydrogen phosphate-dipotassium hydrogen phosphate buffer solution, pH 7.2), let it stand for 40 s, and then discard it. After repeating this 5 times, put the well upside down on the flattened roll paper Pat dry.

[0033] Step 2: Add 100 μL of 15 mg / mL bovine serum albumin (BSA) (prepared in 12 mM sodium bicarbonate-sodium carbonate buffer solution, pH 9.5) to a single well and let stand at room temperature for 5 h. Discard the liquid and shake dry. Then f...

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Abstract

The invention discloses an instrument-free enzyme-linked immunosorbent assay method. According to the method, an antigen-antibody specific immune reaction is carried out in a 96-pore plate so as to capture an analyte antigen or antibody in a sample; nanoparticles are introduced to catalyze a reaction for generating nanogold between chloroauric acid and a reducing agent; a watch or a mobile phone is used for recording the time required for mixed solution to become red, so as to complete instrument-free enzyme-linked immunosorbent assay, wherein the time is negatively correlated to the concentration of a target antigen or antibody in the sample. According to the method, widespread watches or mobile phones are used for carrying out signal reading, so that the cost of quantitative analysis isreduced. Through collaborative use of lipidosome for wrapping plenty of nanoparticles and efficient catalytic activity of the nanoparticles for the reaction of reducing the chloroauric acid by the reducing agent to generate the nanogold, the response signals for the specific binding reaction of single antigen-antibody are amplified, so that the detection sensitivity is improved.

Description

technical field [0001] The invention belongs to the technical field of enzyme-linked immunosorbent assay (Enzyme-Linked Immune-Sorbent Assay, abbreviated as ELISA), and specifically relates to an instrument-free enzyme-linked immunosorbent assay method. Background technique [0002] Enzyme-linked immunosorbent assay (ELISA) is an experimental technique with high specificity and high sensitivity, which is based on immunological reactions and combines the specific reactions of antibodies and antigens with the high-efficiency catalysis of enzymes on substrates. According to the different types and properties of the target to be tested in the sample, various types of ELISA methods can be designed, including antibody sandwich method, antigen sandwich method, competition method, indirect method and capture coating method, etc. Since Engvall and Perlmann of Stockholm University in Sweden first established the enzyme-linked immunosorbent assay method in 1971, after more than 40 year...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53
CPCG01N33/53G01N33/5306
Inventor 张云刘召应聂瑾芳黄锦坤
Owner GUILIN UNIVERSITY OF TECHNOLOGY
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