Immune chromatography test paper detection method based on catalyzing and amplifying of detection signal

An immunochromatographic test paper and signal detection technology, which is applied in biological testing, material inspection products, etc., can solve the problems of detection sensitivity, difficulty in distinguishing detection signals, and fuzzy detection signals, so as to achieve easy judgment and preservation, and broad application prospects , fast effect

Inactive Publication Date: 2013-04-24
SOUTHEAST UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, because the silver staining process of gold standard silver staining technology usually needs to be carried out in a dark room, and the speed of automatic deposition of silver is also very fast, the detection signal is easily blurred by the continued deposition of silver in a short period of time, resulting in serious non-specificity. It is difficult to distinguish signals; directly using magnetic nanomaterials instead of colloidal gold as markers to detect magnetic signals, although the purpose of quantitative detection is achieved, it still cannot solve the problem of low detection sensitivity; fluorescence is easily quenched and unstable

Method used

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  • Immune chromatography test paper detection method based on catalyzing and amplifying of detection signal
  • Immune chromatography test paper detection method based on catalyzing and amplifying of detection signal
  • Immune chromatography test paper detection method based on catalyzing and amplifying of detection signal

Examples

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Embodiment 1

[0039] Example 1: Determination of human chorionic gonadotropin (HCG) by double-antibody sandwich method

[0040] (1) Preparation of nanomaterials: c(trisodium citrate) / c(H 2 PdCl 4 )=20, c(H 2 PdCl 4 )=0.5mM as an example, add 15ml H to a 100ml round bottom flask 2 PdCl 4 solution (2mM), 0.1784g trisodium citrate, 2ml ethanol and 43mlddH 2 O, with mechanical stirring, heated to reflux at 115°C for 3h. The solution turns from yellow to brown yellow and then turns brown, and the Pd sol stabilized by trisodium citrate is obtained.

[0041] (2) Nanomaterial-labeled antibody: The prepared Pd sol was centrifuged at high speed (13000rpm, 10min), the supernatant was removed, concentrated 5 times and then redissolved in 1ml ddH 2 O, adjust to the optimum pH 9.0, add β-HCG monoclonal antibody, incubate at room temperature for one hour, add 100 μL 10% BSA, react for 30 minutes, centrifuge at 4°C×12000rpm×20min to retain the precipitate, and remove the supernatant. Wash the marke...

Embodiment 2

[0052] Embodiment 2: Determination of benzopyrene by competitive method

[0053] (1) Preparation of nanomaterials: first, 0.0355g PdCl 2 Dissolve 47μL of concentrated hydrochloric acid solution (12M) in 100mL of water, heat and ultrasonically disperse to form 2mM H 2 PdCl 4 solution. Next, weigh 0.045g MUA and 0.1g hydrazine hydrate respectively, place them in a three-necked bottle, and add 15mL of water. After the MUA was completely dissolved, its pH was adjusted to 12 with 0.1M NaOH solution, while the temperature of the solution was controlled at 35°C. Then, 5 mL of the 2 mM H 2 PdCl 4 The solution was slowly dripped into the three-necked bottle (speed 1ml / 5min). As the reaction progressed, the color of the solution gradually changed from colorless to black. After the reaction, the above solution was centrifuged at 13000 rpm for 20 minutes, and after removing the supernatant, it was dispersed in water again. Repeat the above process 5 times to remove excess stabiliz...

Embodiment 3

[0059] Embodiment 3: the preparation of nanometer material

[0060] In the above embodiments, we have only selected two kinds of nanomaterials for specific description. In fact, there are many kinds of nanomaterials that can be applied in the present invention, such as:

[0061] a. Preparation of palladium particles or platinum particles:

[0062] (1) Preparation of palladium particles: c (trisodium citrate) / c (H 2 PdCl 4 )=20, c(H 2 PdCl 4 )=0.5mM as an example, add 15ml H to a 100ml round bottom flask 2 PdCl 4 Solution (2mM), 0.1784g trisodium citrate, 2ml ethanol and 43mlddH2O, mechanically stirred, heated to reflux at 115°C for 3h. The solution turns from yellow to brown yellow and then turns brown, and the Pd sol stabilized by trisodium citrate is obtained.

[0063] (2) Preparation of platinum particles: c (trisodium citrate) / c (H 2 PtCl 4 )=20, c(H 2 PtCl 4 )=0.5mM as an example, add 15ml H to a 100ml round bottom flask 2 PtCl 4 Solution (2mM), 0.1784g triso...

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Abstract

The invention discloses an immune chromatography test paper detection method based on catalyzing and amplifying of a detection signal. The immune chromatography test paper detection method comprises the following steps: capturing a molecule to be detected to a detecting site through the chromatographic technique; and catalyzing and amplifying the detection signal under the catalyzing effect. According to the immune chromatography test paper detection method based on catalyzing and amplifying of the detection signal, the biomolecule such as immunoglobulin is marked by a catalytic nanometer material; the detection signal is further amplified through an immune chromatography and a chemical plating technology, so that the sensitivity is improved; a large number of catalytic systems of nano-catalysts are provided, the nano-catalysts can catalyze silver, and also can catalyze the settlement of magnetic materials such as cobalt nickel, so that a magnetic signal can be read while the signal is detected, and as a result, the purposes of quantitative detection and semi-quantitative analysis can be realized.

Description

technical field [0001] The invention belongs to the field of nanometer catalysis and relates to a biological detection method based on immunochromatographic test paper. Background technique [0002] The immunochromatography method appeared in the early 1980s. It is a unique immunoassay method. The core technology of this method is to use the fiber chromatography material as the solid phase, and the sample solution swims on the chromatography material through capillary action. , and at the same time, the analyte labeled with the marker in the sample and the receptor coated on the chromatographic material for the analyte undergo a highly specific and high-affinity immune reaction enriched on the detection band, through the enzymatic reaction Or directly use visual markers to obtain intuitive experimental phenomena. [0003] Immunochromatographic methods are widely used, and can be further divided into classic detection techniques such as enzyme immunolabeling, fluorescent imm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
Inventor 李红英王志飞郑爽何农跃
Owner SOUTHEAST UNIV
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