Time resolution fluorescence detection kit based on phosphorescence luminous technology, and preparation method and application of detection kit

A time-resolved fluorescence and detection kit technology, applied in the field of immunoassay, can solve the problems of affecting sensitivity, increasing operation steps and reaction time, delaying the reporting time, etc., to achieve the effect of sensitive qualitative and quantitative detection and analysis

Inactive Publication Date: 2013-09-11
SHENZHEN AIRUI BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the enhancement solution is easily contaminated during the preparation and use, which will increase the detection background signal and affect the sensitivity of its determination.
In addition, the use of enhanced liquid also increases the operation steps and reaction time during use, which is very inconvenient for the detection of large batches of samples, greatly delaying the report time, and not using clinicians to diagnose and treat the disease in time

Method used

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  • Time resolution fluorescence detection kit based on phosphorescence luminous technology, and preparation method and application of detection kit
  • Time resolution fluorescence detection kit based on phosphorescence luminous technology, and preparation method and application of detection kit
  • Time resolution fluorescence detection kit based on phosphorescence luminous technology, and preparation method and application of detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1: Detection of hepatitis B virus surface antigen HBsAg by double-antibody sandwich method

[0051] The preparation method of the time-resolved fluorescence detection kit for quantitative detection of hepatitis B virus surface antigen comprises the following steps:

[0052] 1. Preparation of anti-HBs labeled with phosphorescent luminescent material:

[0053] Dilute the anti-HBs monoclonal antibody B with 0.1M pH9.6 sodium bicarbonate-sodium carbonate solution to 1mg / ml, take 5ml of antibody solution, add 30mg of phosphorescent material metalloporphyrin solution, stir well, and incubate at room temperature for 1 hour , and mix every 15 minutes. Finally, use a G25 gel column for separation and purification, collect the marked metalloporphyrin-labeled anti-HBs antibody B, and use 1.5% bovine serum albumin, 10% fetal bovine serum, 0.1% preservative Proclin300, 0.2 % protectant diluted with 0.05mol / L pH7.2 Tris-HCl buffer solution, packed in sealed reagent bottles, a...

Embodiment 2

[0075] Example 2: Detection of HIV Antibody by Double Antigen Sandwich Method

[0076] The preparation method of qualitative detection HIV antibody time-resolved fluorescence detection kit comprises the following steps:

[0077] 1. Preparation of HIV antigen labeled with phosphorescent material:

[0078] Dilute HIV antigen B to 1mg / ml with 0.1M pH9.6 sodium bicarbonate-sodium carbonate solution, take 5ml antibody solution, add 30mg phosphorescent material metalloporphyrin solution, stir well, incubate at room temperature for 1 hour, every Mix once every 15 minutes. Finally, use a G25 gel column for separation and purification, collect the marked metalloporphyrin-labeled anti-HIV antigen B, and use 1.5% bovine serum albumin, 10% fetal bovine serum, 0.1% preservative Proclin300, 0.2 % protectant diluted with 0.05mol / L pH7.2 Tris-HCl buffer solution, packed in sealed reagent bottles, and stored at 2-8°C.

[0079] 2. Preparation of magnetic particles coupled with HIV antigen A:...

Embodiment 3

[0095] Embodiment 3: competition method mode detects morphine

[0096] The preparation method of quantitative detection morphine immunochromatography kit comprises the following steps:

[0097] 1. Preparation of anti-morphine antibody labeled with phosphorescent material:

[0098] Dilute rabbit anti-morphine monoclonal antibody B to 1mg / ml with 0.1M pH9.6 sodium bicarbonate-sodium carbonate solution, take 5ml antibody solution, add 30mg phosphorescent material metalloporphyrin solution, stir well, and incubate at room temperature for 1 Hours, mix every 15 minutes. Finally, use a G25 gel column for separation and purification, collect the labeled metalloporphyrin-labeled morphine antibody B, and use 1.5% bovine serum albumin, 10% fetal bovine serum, 0.1% preservative Proclin300, 0.2% The protective agent is diluted with 0.05mol / L pH7.2 Tris-HCl buffer solution, sealed and packaged in a reagent bottle, and stored at 2-8°C.

[0099] 2. Preparation of BSA-morphine coupling comp...

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Abstract

The invention provides a time resolution fluorescence detection kit based on a phosphorescence luminous technology, and a preparation method and application of the detection kit. The kit comprises magnetic particles covered by an antigen or an antibody, an antigen or an antibody marked with a phosphorescence luminous material, a standard product of an object to be detected, a washing solution and a micro-pore plate. The kit adopts the phosphorescence luminous material, namely platinum / palladium porphyrin, as a biological marker. The preparation method of the kit comprises the following steps of: preparing the magnetic particles covered by the antigen or the antibody, marking the antigen or the antibody by the phosphorescence luminous material, preparing the standard product of the object to be detected, and preparing the washing solution. The invention further discloses the application of the kit to quantitative determination of a biological sample. The kit is divided into a sandwich method mode, a competition method mode, an indirection method mode and a capturing method mode according to different immune reaction manners of objects to be detected; rapid and sensitive qualitative and quantitative detection analysis on the different objects to be detected in samples can be carried out by different detection modes according to different properties of the objects to be detected.

Description

technical field [0001] The invention relates to a time-resolved fluorescence detection kit based on phosphorescence technology and a preparation method and application thereof, belonging to the technical field of immune detection. Background technique [0002] Immunoassay is based on the immune binding reaction between antigen and corresponding antibody, that is, using antibody (or antigen) as a selective reagent to determine antigen, hapten (or antibody). The reaction between antibody and antigen has extremely high specificity, so no or only simple pretreatment is required for immunoassay samples. Antibody-antigen complexes have high stability, and their stability constants are generally 10 9 , even up to 10 15 . Unfortunately, although antibody-antigen binding can display certain catalytic properties, from the perspective of analytical testing, the immune response lacks the analytical signal contrast of using highly sensitive reagents, making it difficult to detect dire...

Claims

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Application Information

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IPC IPC(8): G01N33/543
Inventor 谢爱武
Owner SHENZHEN AIRUI BIO TECH
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