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Kit for quantitatively detecting GFAP and application of kit

A technology for detecting reagents and kits, applied in the field of kits for quantitative detection of GFAP, which can solve the problems of poor linearity, poor fluidity, poor accuracy and stability of test results, etc., so that it is not easy to miss judgments and reduce misjudgments and missed judgment, the effect of strong objectivity

Active Publication Date: 2021-03-12
SOPHONIX CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The invention improves the signal intensity and sensitivity of the immune response, enables low-content substances to generate strong chemiluminescence signals when immunocombined, and provides a more accurate, Accurate, convenient, fast and simple method, however, the glial fibrillary acidic protein (GFAP) antibody labeled with magnetic particles used in this invention is easy to aggregate and precipitate, has poor fluidity, and is prone to residues on the wall during use, so that it can be used during the detection process. The number of active particles is reduced, resulting in poor accuracy and stability of test results
[0008] CN 105300966A discloses a preservation solution, including 0.1-10g / L buffer salt, 100-300g / L antibody stabilizer, 0.1-1g / L antiseptic and antibacterial agent, 2-12g / L carrageenan and water. The buffer salt, antibody stabilizer and antiseptic and antibacterial agent in the solution work together to ensure the stability of the magnetic microparticles or magnetic nanoparticles and antibodies in the whole system, but this system leads to poor linearity of the kit detection and accurate detection results Poor performance and stability, unable to meet industry standards

Method used

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  • Kit for quantitatively detecting GFAP and application of kit
  • Kit for quantitatively detecting GFAP and application of kit
  • Kit for quantitatively detecting GFAP and application of kit

Examples

Experimental program
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Effect test

preparation example Construction

[0059] The preparation method of the reagent A is as follows: dissolving the enzyme-labeled GFAP antibody conjugate in buffer 8 and mixing uniformly; the buffer 8 consists of the following components: Na 2 HPO 4 12H 2 O 5.6-5.9g / L, NaCl 9.0g / L and bovine serum albumin 1.0-50g / L, pH 7.0-7.6.

[0060] The preparation method of the enzyme-labeled GFAP antibody conjugate is: comprising the following steps:

[0061] B1. Antibody activation:

[0062] B1.1 Activation step: Weigh 4-8mg of 2-iminosulfane hydrochloride (2-IT), dissolve it in buffer 1 to 13.76mg / mL, and set the molar ratio of 2-IT to antibody at 15-30:1 Add the 2IT solution to the antibody solution for activation (i.e. add 1mg antibody to 10-20μL of 2IT solution), shake and mix well, and react at room temperature for 30 minutes to obtain the activated antibody pre-product;

[0063] B1.2 Termination of activation step: add buffer 2 to the activated antibody pre-product solution at the ratio of 1 mg antibody to 5-20 μL...

Embodiment 1

[0093] A kit for quantitative detection of GFAP consists of a detection reagent strip, a calibrator, a quality control product and a two-dimensional code (as shown in Table 1), wherein the detection reagent strip is composed of a series of solutions and accessories as a whole, which can be independently detected One sample; the calibrator is prepared from two concentrations of GFAP antigen and buffer to calibrate the standard curve; the quality control is prepared from GFAP antigen and buffer; the standard curve of the current batch is entered in the QR code .

[0094] Table 1 Component list of the kit

[0095]

[0096]

[0097] The detection reagent strip includes reagent A, reagent B, cleaning solution, luminescent substrate, measuring and reading hole, elution sleeve and suction tip, wherein, reagent A is a GFAP antibody solution containing a certain concentration of alkaline phosphatase labeled; reagent B is a GFAP antibody solution containing a certain concentratio...

Embodiment 2

[0125] A kit for quantitatively detecting GFAP consists of a detection reagent strip, a calibrator, a quality control product and a two-dimensional code (as shown in Table 2), wherein the detection reagent strip is composed of a series of solutions and accessories as a whole, which can be independently detected One sample; the calibrator is prepared from two concentrations of GFAP antigen and buffer to calibrate the standard curve; the quality control is prepared from GFAP antigen and buffer; the standard curve of the current batch is entered in the QR code .

[0126] Table 2 Component list of the kit

[0127] The main components of the kit quantity Detection Reagent Strips 10 pieces Quality Control 200μL×1 Calibrator 1 200μL×1 Calibrator 2 200μL×1 Box label QR code 1

[0128] The detection reagent strip includes reagent A, reagent B, cleaning solution, luminescent substrate, measuring and reading hole, elution sleeve and suc...

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Abstract

The invention discloses a kit for quantitatively detecting GFAP and application of the kit, the kit comprises a magnetic particle labeled GFAP antibody solution, the buffer solution comprises the following components: 5.6-5.9 g / L of Na2HPO4. 12H2O, 0.55-0.60 g / L of NaH2PO4, 9.0 g / L of NaCl, 1.0-50 g / L of bovine serum albumin, 80.0-140 g / L of sucrose, 1.0-5.0 g / L of cellulose salt or cellulose derivative and 5.0-50 g / L of gelatin, wherein the pH value is 6.2-8.0. By improving a buffer system of a magnetic particle carrier in a magnetic particle luminescence method, the fluidity and sedimentation performance of the magnetic particle antibody conjugate are improved, the accuracy and repeatability of the kit are improved, the stability is high, meanwhile, the detection limit is low, the linearrelation is good, and the industrial standard is met.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a kit for quantitatively detecting GFAP and an application thereof. Background technique [0002] Traumatic brain injury (TBI) is an injury to the brain caused by an external force that disrupts normal brain function, resulting in impairment of a person's cognitive abilities or physical functions. Among all types of TBI, the most common sequelae were headache (47.9%) and abnormal memory (42%), and about 30% of patients needed psychological counseling or neurological treatment. TBI is the most common disease in neurosurgery, and it is also one of the main causes of death and disability worldwide, and its sequelae have permanent impact on patients' health. Studies have shown that the incidence of TBI worldwide is 790-979 per 100,000 per year, and an estimated 54-60 million people are newly diagnosed with TBI each year. Based on this estimate, there are 11...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543G01N33/535
CPCG01N33/535G01N33/54326G01N33/6893G01N2333/47G01N2800/28G01N2800/40
Inventor 刘聪郑兴华李博飞冯玉静
Owner SOPHONIX CO LTD
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