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Electrochemiluminescence multicomponent immunodetection method based on spectral resolution principle

An immunoassay method and electrochemistry technology, applied in the field of electrochemiluminescence multi-component immunoassay, can solve the problems that ECL analysis technology is still under development, consume a lot of reagents and samples, and take a long time to achieve monochromaticity Good results with less sample consumption and less work

Active Publication Date: 2016-11-16
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This analysis mode takes a long time, consumes a lot of reagents and samples, and is not an ideal analysis mode
[0004] The ECL multi-component immunoassay research based on the principle of spectral resolution is still blank, and the related ECL analysis technology is still under development

Method used

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  • Electrochemiluminescence multicomponent immunodetection method based on spectral resolution principle
  • Electrochemiluminescence multicomponent immunodetection method based on spectral resolution principle
  • Electrochemiluminescence multicomponent immunodetection method based on spectral resolution principle

Examples

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Embodiment 1

[0075] An electrochemiluminescent multi-component immunoassay method based on the principle of spectral resolution, comprising the following steps:

[0076] (1) Preparation of corresponding secondary antibodies labeled with three kinds of quantum dots

[0077] Add 10 μL phosphate buffer solution containing 100 mg / mL 1-ethyl-3,3-dimethylaminopropylcarbodiimide and 10 μL containing 100mg / mL N-hydroxysuccinimide phosphate buffer solution, react at room temperature for 30min; after centrifugation and purification at 12000rpm, take the precipitate and add 100μL phosphate buffer solution for ultrasonic dispersion, take 15μL of the above-mentioned activated CdSe quantum dot solution and add 150 μL of 100 μg / mL carcinoembryonic antigen secondary antibody, shake at room temperature for 3 hours; then add 20 μL of 1% volume concentration of bovine serum albumin, incubate at room temperature for 1 hour, centrifuge and purify at 12000 rpm, take the precipitate and add 100 μL of phosphate b...

Embodiment 2

[0088] A multi-component immunoassay method based on electrochemiluminescence, the steps are the same as in Example 1, except that:

[0089] Carcinoembryonic antigen, prostate specific antigen, and alpha-fetoprotein antigen were replaced by carbohydrate antigen 125, carbohydrate antigen 15-3, and carbohydrate antigen 72-4, respectively.

Embodiment 3

[0091] A multi-component immunoassay method based on electrochemiluminescence, the steps are the same as in Example 1, except that:

[0092] Carcinoembryonic antigen, prostate specific antigen, and alpha-fetoprotein antigen were replaced by carbohydrate antigen 19-9, free prostate specific antigen, and AIDS antigen, respectively.

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Abstract

The present invention relates to an electrochemiluminescence multicomponent immunodetection method based on a spectral resolution principle, wherein CdSe quantum dots and CdTe quantum dots are adopted as markers, and the ECL signals of different markers are identified based on a spectral resolution principle so as to provide the ECL multicomponent detection method. The method mainly comprises: (1) preparing three quantum dot labeled corresponding secondary antibody (QDs-Ab2); (2) preparing a multicomponent ECL immunosensor using the three quantum dots as markers; and (3) carrying out multicomponent ECL immunodetection. According to the present invention, the detection sensitivity of the method is high, the detection limit of the carcinoembryonic antigen achieves 1.0 pg / mL, the detection limit of the prostate specific antigen achieves 10.0 pg / mL, the detection limit of the alpha fetoprotein antigen achieves 10.0 fg / mL, the concurrent detection of a variety of antigens can be achieved, and the selectivity is high.

Description

technical field [0001] The present invention relates to an electrogenerated chemiluminescence (ECL) multi-component immunoassay method, in particular to a multi-component immunoassay using mercaptopropionic acid and sodium hexametaphosphate-coated CdSe quantum dots and CdTe quantum dots as markers. Component immunoassay method. Background technique [0002] Electrogenerated chemiluminescence (ECL) is an analytical technique that has been widely used in the fields of biochemical analysis and clinical diagnosis. For example, the kit based on the ruthenium bipyridine / n-tripropylamine ECL system developed by Roche has been used in the clinical diagnosis of more than 80 items such as tumor markers and hormones. Studies have shown that quantum dots have excellent photoelectric properties, and it is possible to develop new ECL biochemical analysis and clinical diagnostic technologies (see: Analyst, 2013, 138, 43-61). For example, Chinese patent document CN102749452A (ZL 201210262...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/58G01N33/543
CPCG01N21/76G01N33/54366G01N33/588
Inventor 邹桂征周洁
Owner SHANDONG UNIV
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