Luminescent substrate, use of luminescent substrate and detection kit containing luminescent substrate

A technology of luminescent substrates and kits, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of unsatisfactory sensitivity and specificity, and achieve improved sensitivity and specificity, strong optical signals, and stability Good results

Inactive Publication Date: 2014-02-26
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, AFP is widely used clinically as a diagnostic marker for liver cancer, but its sensitivity and specificity are not ideal for the diagnosis of early liver cancer

Method used

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  • Luminescent substrate, use of luminescent substrate and detection kit containing luminescent substrate
  • Luminescent substrate, use of luminescent substrate and detection kit containing luminescent substrate
  • Luminescent substrate, use of luminescent substrate and detection kit containing luminescent substrate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The preparation of embodiment 1 chemiluminescence substrate liquid

[0023] The substrate formulation is as follows:

[0024] Recipe 1:

[0025] Luminescent agent solution A liquid: 2.5mmol / L luminol, 3.0mmol / L p-iodophenol, Tris-HCl buffer solution with pH 8.5;

[0026] Oxidant solution B solution: carbamide peroxide 1.3mmol / L, Tris-HCl buffer solution with pH 8.5;

[0027] Recipe 2:

[0028] Luminescent agent solution A: luminol 0.5mmol / L, p-iodophenol 5.0mmol / L, Tris-HCl buffer solution with pH 8.5;

[0029] Oxidant solution B liquid: carbamide peroxide 1mmol / L, Tris-HCl buffer solution with pH 8.5;

[0030] Recipe 3:

[0031] Luminescent agent solution A liquid: luminol 8mmol / L, p-iodophenol 10mmol / L, Tris-HCl buffer solution with pH 8.5;

[0032] Oxidant solution B liquid: carbamide peroxide 10mmol / L, Tris-HCl buffer solution with pH 8.5;

[0033] Recipe 4:

[0034] Luminescent agent solution A liquid: luminol 5mmol / L, p-iodophenol 0.3mmol / L, Tris-HCl buffe...

Embodiment 2

[0042]The kit provided by the present invention includes a microwell plate coated with monoclonal antibody, HRP-labeled monoclonal antibody, freeze-dried standard substance, sample diluent, concentrated washing solution and the chemiluminescence base of any one of the formulations in Example 1. thing.

[0043] The GP73 chemiluminescence quantitative detection kit is used as an example below to illustrate the preparation method of the detection kit, which can also be applied to the detection of other proteins.

[0044] 1. Preparation of solid-phase coated plate

[0045] a) Coating: GP73 monoclonal antibody 5F10 was coated with a concentration of 2ug / ml on a microwell plate, 100ul / well, and coated at 4°C for 18h;

[0046] b) Blocking: wash the plate twice, pat dry, add blocking solution 200ul / well, 37°C for 1.5h.

[0047] c) Seal the bag: Discard the blocking solution, pat dry, dry in a 37°C incubator for 1 hour, and immediately vacuum seal the bag.

[0048] 2. Preparation of...

Embodiment 3

[0053] Embodiment 3 The using method of kit of the present invention

[0054] The specific operations of the GP73 chemiluminescent quantitative detection kit prepared in Example 2 are as follows:

[0055] 1. Reagent and sample preparation

[0056] a) Take out the kit from 4°C, and place it at room temperature for 20-30 minutes;

[0057] b) Add 500ul of pure water to each tube of freeze-dried standard substance to fully dissolve;

[0058] c) Dilute the concentrated washing solution with pure water at 1 / 20 to make 1× washing solution.

[0059] d) Before use, put the sample to be tested at room temperature to balance for 20-30 minutes, shake gently to mix well, if there is sediment in the sample, it should be centrifuged to remove the precipitate before use, the sample used for detection should meet the detection range of the kit, if the concentration is too high , can be diluted with sample diluent before detection.

[0060] 2. Operation steps

[0061] a) Take out the coate...

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Abstract

The invention discloses a luminescent substrate, a use of the luminescent substrate and a detection kit containing the luminescent substrate. The luminescent substrate has high sensitivity and high stability. The luminescent substrate comprises a solution A and a solution B. The solution A comprises 0.5-8mmol / L of luminal, 0.5-10mmol / L of p-lodophenol, and 0.1mol / L of a Tris-HCl buffer solution. The solution B comprises 1-10mmol / L of urea peroxide and 0.1mol / L of the Tris-HCl buffer solution. The kit comprises a monoclonal antibody-coated microporous plate, HRP-labeled monoclonal antibodies, a freeze-dried standard, a sample diluent, a concentrated washing liquid and the chemiluminescent substrate.

Description

technical field [0001] The invention discloses a luminescent substrate, and also discloses the application of the luminescent substrate and a detection kit containing the luminescent substrate. Background technique [0002] In the past ten years, the research and application of marker immunoassay technology has developed rapidly, and has been widely used in various fields of biomedical basic theory research and clinical disease diagnosis. The methods used to detect serological indicators mainly include radioisotope immunoassay, enzyme-linked immunosorbent assay and chemiluminescence immunoassay. [0003] At present, the most widely used quantitative immunoassay method in the clinical field is mainly chemiluminescence immunoassay (CLIA), which is a sensitive micro-immunoassay method, which combines high sensitivity of radioimmunoassay (RIA) and easy operation of enzyme-linked immunoassay (ELISA), etc. Advantages, while overcoming the respective shortcomings of radioimmunoass...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/68
CPCG01N33/543G01N21/763G01N33/57423G01N33/6803
Inventor 彭涛龙飞苏文瀚肖静殷仙丽卢然如
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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