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Chemiluminescent enhancing solution, chemiluminescent substrate and applications of chemiluminescent enhancing solution

A technology of chemiluminescence and enhancement solution, which is applied in the fields of chemiluminescence/bioluminescence, analysis through chemical reaction of materials, and biological testing, etc. It can solve the problem of stable luminol luminescence signal, poor stability of luminescence signal, and complex reaction system issues such as chemicalization, to achieve the effects of good luminescent signal stability, improved detection accuracy, and good storage stability

Active Publication Date: 2018-08-03
LEADWAY HK
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this requires the addition of an additional hydrolase, which complicates the reaction system and increases the detection cost
Furthermore, although the luminescence signal of luminol can be improved by only using o-phenylphenol as an enhancer, it still leads to problems such as high background signal and poor stability of luminescence signal in the HRP-luminol chemiluminescence system.
In addition, due to some unknown reason, o-phenylphenol itself makes it difficult for the enhanced luminol luminescence signal to remain stable for a long period of time, and the luminescence signal decays relatively quickly

Method used

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  • Chemiluminescent enhancing solution, chemiluminescent substrate and applications of chemiluminescent enhancing solution
  • Chemiluminescent enhancing solution, chemiluminescent substrate and applications of chemiluminescent enhancing solution
  • Chemiluminescent enhancing solution, chemiluminescent substrate and applications of chemiluminescent enhancing solution

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Experimental program
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Effect test

Embodiment 1

[0055] Embodiment 1 Chemiluminescence reagent and detection method thereof

[0056] In the present invention, the analyte is an antigen, an antibody or a hapten, and the antigen AFP is used as an example for illustration. The solid phase can be selected from microporous plates, plastic microparticles, magnetic beads, elastic plastic tubes or plastic beads, and a microporous plate is used as an example for illustration.

[0057] 1. HRP marker preparation method

[0058] HRP markers refer to HRR-labeled antigens, antibodies or haptens. Commonly used labeling methods include glutaraldehyde method and sodium periodate oxidation method. Here, the sodium periodate oxidation method is taken as an example for illustration. The detailed steps are:

[0059] (1) Weigh 5 mg of HRP and dissolve it in 1 ml of distilled water to form a HRP solution with a concentration of 5 mg / ml;

[0060] (2) Add 0.2ml of newly prepared 0.1M NaIO to the HRP solution 4 solution, stirred at room temperat...

Embodiment 2

[0091] Embodiment 2: Chemiluminescence substrate experiment

[0092] (1) Chemical enhancer experiment

[0093] The chemiluminescence substrate consists of liquid A and liquid B. In liquid A, buffer: carbonate buffer with pH 9.0±0.1, luminol (LS): 0.6g / L; in liquid B, carbamide peroxide (UP): 0.3g / L, dilute The solution was 0.2M phosphate buffer (pH 7.4±0.1). By changing the o-phenylphenol (D) concentration and the 4-imidazolephenol (E) concentration in A liquid, detect three parameters according to the embodiment 1: reaction system background signal, positive signal value and linear width, wherein reaction The background signal of the system, that is, the blank luminescence value, refers to the luminescence signal generated by detection by adding only the chemiluminescent substrate without adding reagents such as the sample to be tested in the reaction well; the positive signal value, here, the AFP with a concentration of 500ng / mL The calibrator (AFP500) as an example refer...

Embodiment 3

[0119] Example 3: Chemiluminescence substrate storage stability experiment

[0120] In order to test the stability of the chemiluminescent substrate of the present invention, liquid A was prepared: o-phenylphenol (D) 0.04g / L, 4-imidazole phenol (E) 0.125g / L, luminol 0.8g / L, buffer Solution: 0.1M carbonate buffer solution (pH is 9.0±0.1); B solution: carbamide peroxide (UP) 0.3g / L, diluent is 0.2M phosphate buffer solution (pH is 7.4±0.1), according to the implementation For the detection method in Example 1, the luminescent signal intensity (RLU) of the 5 different concentrations of AFP calibrator prepared in Example 1 was detected at a reagent storage condition of 2-8°C at different times, and the detection results are shown in Table 7. Show.

[0121] Table 7. Test results of AFP calibrator

[0122] 2~8℃(month)

[0123] It can be seen from Table 7 that the chemiluminescent substrate of the present invention still maintains a very good correlation coefficient at 2-...

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Abstract

The present invention provides a chemiluminescent enhancing solution, a chemiluminescent substrate, a detection method and a kit for detecting a to-be-detected substance in a sample. According to thepresent invention, the improvement comprises the combined use of two chemiluminescent enhancing agents such as o-phenylphenol and 4-imidazole phenol, wherein the two enhancing agents provide the synergistically-enhanced effect, can significantly reduce the background signal, and can further maintain the stable luminol luminescence signal for a long time; and the detection method and the kit can bematched with the chemiluminescence analyzer, and can be used for detecting tumor markers, infectious diseases, hormones and other enzymatic chemiluminescence products.

Description

technical field [0001] The present invention relates to a chemiluminescence enhancement solution, a chemiluminescence substrate, a detection method and a kit for detecting analytes in samples, all of which involve the combined use of two chemiluminescence enhancers, o-phenylphenol and 4-imidazole phenol, belonging to Immunotechnical field. Background technique [0002] Since the advent of chemiluminescent immunoassay (CLIA) in the 1970s and 1980s, it has excellent specificity and The significantly improved sensitivity has been widely used in the fields of drug analysis, environmental analysis, food analysis and biomedicine, such as the detection of tumor biomarkers in the field of biomedicine, such as alpha-fetoprotein (AFP), cancer Embryonic antigen (CEA), ferritin, prostate specific antigen (PSA), neuron-specific enolase (NSE), CYFRA21-1, CA19-9, CA50, CA125, CA153, CA724, etc.; gonad biomarkers, Such as follicle stimulating hormone (FSH), luteinizing hormone (LH), prola...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76G01N33/53
CPCG01N21/763G01N33/5306
Inventor 毛丽敏吴炜汤海霞
Owner LEADWAY HK
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