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Chemiluminescence immunoassay kit for detecting neuronspecific enolase

A technology of chemiluminescence immunity and enolase, which is applied in chemiluminescence/bioluminescence, biological testing, and analysis through chemical reactions of materials, etc., can solve the problems of narrow linear range of kits and differences in measured values, and achieve sensitivity High, wide linear range, good specificity

Active Publication Date: 2019-01-11
DIRUI MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to solve the problem that the existing test kits for testing NSE have a narrow linear range, and there are significant differences in the measured values ​​of serum and plasma samples, and provide a chemiluminescence method for detecting neuron-specific enolase Immunization Kit

Method used

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  • Chemiluminescence immunoassay kit for detecting neuronspecific enolase
  • Chemiluminescence immunoassay kit for detecting neuronspecific enolase
  • Chemiluminescence immunoassay kit for detecting neuronspecific enolase

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preparation example Construction

[0047] According to the present invention, the preparation method of the kit A includes:

[0048] Step 1: R a 1 Preparation of reaction solution

[0049] Mix buffers and preservatives to make R a 1 base buffer;

[0050] Take commercially available streptavidin magnetic beads (selected from JSR Company, model MS300 / Streptavidin), the magnetic beads have a particle size of 3 μm, wash with PBS (20mmol / L PB and 150mmol / L NaCl) buffer solution with pH 7.2 three times, then resuspend the beads to R a 1 In the basic buffer solution, make R with a mass concentration of 0.03%-0.1% a 1 reaction solution;

[0051] Step 2: R a 2 Preparation of reaction solution

[0052] Mix NaCl, magnesium chloride, buffer, preservative, blocking agent, protein stabilizer and surfactant to make R a 2 base buffer;

[0053] Take the NSE antibody and label it with the acridinium ester solution dissolved in the solvent according to the ratio of molar concentration 1:(1-20). The labeling buffer is pH7...

Embodiment 1

[0115] Embodiment 1 Kit A

[0116] 1.1 Kit composition

[0117]

[0118] 1.2 Preparation method of the kit

[0119] (1)R a 1 Preparation of reaction solution

[0120] Take streptavidin magnetic beads with a particle size of 3 μm, wash them three times with PBS buffer at pH 7.2, and then resuspend the magnetic beads to an appropriate amount of R a 1 In the basic buffer solution (other components except magnetic beads), make R with a mass concentration of 0.1% a 1 reaction solution.

[0121] (2) R a 2 Preparation of reaction solution

[0122] a. Preparation of acridinium ester-labeled NSE antibody:

[0123] Take an appropriate amount of NSE antibody, and label it with acridinium ester (3mg / mL) dissolved in DMF according to the molar concentration ratio of 1:7.5. The labeling buffer is pH7.4PB (20mmol / L), and the labeling time is 4h. The reaction process requires Avoid light. After the labeling reaction, use the AKTA purification instrument (G25 gel column) to purify,...

Embodiment 2

[0133] Embodiment 2 Kit A

[0134] 1.1 Kit composition

[0135]

[0136] 1.2 Preparation method of the kit

[0137] (1)R a 1 Preparation of reaction solution

[0138] Take streptavidin magnetic beads with a particle size of 3 μm, wash them three times with PBS buffer at pH 7.2, and then resuspend the magnetic beads to an appropriate amount of R a 1 In the basic buffer solution (other components except magnetic beads), make R with a mass concentration of 0.1% a 1 reaction solution.

[0139] (2) R a 2 Preparation of reaction solution

[0140] a. Preparation of acridinium ester-labeled NSE antibody:

[0141] Take an appropriate amount of NSE antibody, and label it with acridinium ester (3mg / mL) dissolved in DMF according to the molar concentration ratio of 1:3. The labeling buffer is pH7.4PB (20mmol / L), and the labeling time is 12h. The reaction process requires Avoid light. After the labeling reaction, use the AKTA purification instrument (G25 gel column) to purify, ...

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Abstract

The invention provides a chemiluminescence immunoassay kit for detecting neuronspecific enolase, and belongs to the technical field of in vitro diagnosis. The kit A comprises a calibrator, a magneticparticle reaction liquid Ra1, a trace binder reaction liquid Ra2 and a capture antibody reaction liquid Ra3. The kit B comprises a calibrator, a magnetic particle reaction liquid Rb1, a trace binder reaction liquid Rb2 and a project dilution reaction liquid Rb3. The kit 3 comprises a calibrator, a magnetic particle reaction liquid Rc1 and a trace binder reaction liquid Rc2. The kit has wide detection linear range, high sensitivity and good specificity, is an automatic chemiluminescence instrument mated reagent, and is applicable to quantitative detection of NSE in serum and plasma. According to the kit, consistent detection on serum and EDTA plasma samples can be realized by adding a proper amount of Mg<2+> ions, and the serum and plasma measured value related coefficient r is more than 0.975.

Description

technical field [0001] The invention belongs to the technical field of in vitro diagnosis and relates to a chemiluminescent immunoassay kit for detecting neuron-specific enolase. Background technique [0002] Neuron-specific enolase (NSE) is one of the enolases involved in catalyzing the conversion of 2-phosphoglycerate into phosphoenolpyruvate during glycolysis, with a molecular weight of 78KD and an isoelectric point of pH4 .7, is an acid protease. Enolase isoenzyme is composed of three subunits of α, β, and γ in the form of dimers, and there are five forms of αα, αβ, ββ, αγ, and γγ, of which the α subunit mainly exists in the liver, Kidney and other tissues, dimer αα is called non-neuron-specific enolase, β subunit is unique to skeletal muscle and cardiac muscle, γ subunit is unique to nervous tissue and neuroendocrine tissue, and dimer γγ is in nerve tissue Dimer αγ is expressed in microglia, oligodendrocytes and astrocytes, so dimer αγ and γγ are called NSE. NSE simu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/573G01N21/76G01N33/532G01N33/543G01N33/577G01N33/58
CPCG01N21/76G01N33/532G01N33/54326G01N33/573G01N33/577G01N33/58G01N2800/28G01N2800/52
Inventor 孟令敏李磊王凯孙成艳高威何浩会
Owner DIRUI MEDICAL TECH CO LTD
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