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Chemoluminescent substrate

A technology of chemiluminescence and substrate solution, which is applied in the field of immunoassay, can solve the problems of low sensitivity and limited effect of enhancing luminescence, and achieve the effect of low cost

Active Publication Date: 2009-11-04
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with the formula using only p-iodophenol as the enhancer, the effect of enhancing luminescence is limited, and the sensitivity is still low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Prepare luminescent agent solution and oxidizing agent solution of 1000mL chemiluminescence substrate solution, the specific formula is as follows.

[0033] Chemiluminescent substrate solution Luminescent agent solution:

[0034] Luminol 10mmol / L

[0035] 4-Hydroxybiphenyl 0.3mmol / L

[0036] 4-iodophenylboronic acid 0.05mmol / L

[0037] 0.2mol / L boric acid-borax buffer pH 8.7

[0038] Chemiluminescence substrate solution oxidant solution:

[0039] Carbamide peroxide 3.5mmol / L

[0040] Tween 20 0.1% (volume)

[0041] 0.2mol / L phosphate buffer pH7.2

[0042] The preparation of 1000mL luminescent agent solution of chemiluminescence substrate can adopt the following method: take 1.7716g of luminol, 0.051g of 4-hydroxybiphenyl, 0.012g of 4-iodophenylboronic acid, 11.4g of boric acid, 4.9g of borax, and use bis Dissolve in distilled water to 1000mL, pH 8.7.

[0043] The preparation of 1000mL oxidant solution of chemiluminescent substrate solution can adopt the followin...

Embodiment 2

[0045] Compare the luminescent intensity of the luminescent substrate solution described in the above-mentioned Example 1 with the signal Reagent A and signal Reagent B luminescent substrate solutions imported from the United States monobind company in the assay for the determination of thyroid stimulating hormone (TSH).

[0046] Experimental procedure: add 50 μL of sample, calibrator and enzyme marker to a microwell plate, react at room temperature for 45 minutes, wash 5 times with washing solution, then dry it on absorbent paper, add the mixed luminescent substrate to each well After 100 μL of the substance solution was protected from light at room temperature for 5 minutes, the relative luminescence value (RLU) of each well was detected with a chemiluminescence instrument.

[0047] The commercially available signal Reagent A and signal Reagent B luminescent substrate solutions produced by the monobind company in the United States and the luminescent substrate solution in Exa...

Embodiment 3

[0068] Compare the luminescent substrate solution described in the above-mentioned Example 1 with the commercially available signal Reagent A and signal Reagent B luminescent substrate solutions (imported control luminescent substrate solution) produced by the U.S. monobind company to measure the thyroid-stimulating hormone (TSH) in the experiment. Luminous plateau.

[0069] Add 50 μL of each sample and enzyme marker to a microwell plate, react at room temperature for 45 minutes, wash 5 times with washing solution, then dry it on absorbent paper, and add 100 μL of mixed luminescent substrate solution to each well. After adding the luminescent substrate solution, measure it with a luminometer every 5 minutes for 70 minutes, and count the changes in the luminescence value of the same sample within 70 minutes.

[0070] The imported luminescent substrate solution and the luminescent substrate solution described in Example 1 were used for experiments respectively.

[0071] The res...

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Abstract

The invention discloses a chemoluminescent substrate, which can be applied in the field of immunological detections, and comprises luminescent agent solution and oxidant solution, wherein the luminescence agent solution contains reinforcers, namely 4-hydroxydiphenyl and 4-iodo-borophenylic acid, a luminescent agent, namely luminal or luminal derivatives and buffer solution. The chemoluminescent substrate has high stability, good luminescence effect, long luminescent time, high sensitivity and low cost.

Description

technical field [0001] The invention relates to the field of immunoassay, and more specifically relates to a chemiluminescent substrate liquid applicable to the field of immunoassay. Background technique [0002] Chemiluminescence refers to the emission of light accompanying the chemical reaction process. When certain substances undergo a chemical reaction, they absorb the chemical energy generated during the reaction, and the reaction product molecules are excited to an electronically excited state. When electrons return from the lowest vibrational energy level of the excited state to the various vibrational energy levels of the ground state, radiation is produced, and the excess energy is released in the form of photons. This phenomenon is called chemiluminescence. Luminescence immunoassay (LIA) is a new type of labeled immunoassay technology that combines chemiluminescence or bioluminescence system with immune reaction to detect trace antigen or antibody. LIA has the ad...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/52
Inventor 张黎应希堂宋胜利胡国茂郑金来唐宝军于尚永
Owner CHEMCLIN DIAGNOSTICS CO LTD
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