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Luteinizing hormone (LH) quantitative determination kit, and making method and detection method thereof

A technology for the quantitative determination of luteinizing hormone, which is applied in the direction of chemiluminescence/bioluminescence, and analysis by making materials undergo chemical reactions, can solve the problems of unfavorable high-throughput automatic detection, poor specificity, and low sensitivity. Achieve the effect of optimizing the chemiluminescence enhancement system, small variation and high sensitivity

Inactive Publication Date: 2016-03-30
JIANGSU ZECEN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This technique has low sensitivity, narrow linearity, poor specificity, and is not conducive to high-throughput automatic detection

Method used

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  • Luteinizing hormone (LH) quantitative determination kit, and making method and detection method thereof
  • Luteinizing hormone (LH) quantitative determination kit, and making method and detection method thereof
  • Luteinizing hormone (LH) quantitative determination kit, and making method and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] The configuration of various buffers is as follows:

[0055] 1. Tris salt pH8.0 buffer

[0056] Tris: 12.12g, sodium chloride 5.82g, add to 1L of purified water, stir well until completely dissolved, adjust the final pH to 7.5 with hydrochloric acid.

[0057] 2. Preparation of calibrator buffer

[0058] Add 0.01g of tetracycline and 0.1g of neomycin sulfate to 1L of healthy male serum, fully dissolve and process through 0.22μm filter membrane to obtain.

[0059] 3. Anti-reagent buffer

[0060] Tris: 12.12mg~60.57mg, tetracycline: 0.01g~0.05g, sheep serum: 1g~5g, newborn bovine serum 3g~10g, horse serum 1g~5g, add 1L purified water, stir well until completely dissolved;

[0061] 4. Magnetic particle buffer

[0062] Tris: 12.12mg, sodium chloride 5.82mg, methyl cellulose ether 50g, add to 1L of purified water, stir well until completely dissolved.

[0063] 5. Luminescence substrate buffer

[0064] Tris12.12g~121.14g, sodium chloride 5.82g, lucigenin 0.03g, add to 1L...

Embodiment 2

[0067] Example 2: Preparation of the Quantitative Assay Kit for Luteinizing Hormone LH

[0068] 1. Preparation of calibrators and quality controls

[0069] First: Dissolve luteinizing hormone LH with standard buffer solution, and prepare calibration products and quality control products with target concentrations as shown in Table 1; the luteinizing hormone LH used in this example is purchased from Fitzgerald, a well-known manufacturer at home and abroad .

[0070] Table 1: Preparation of calibrators and controls

[0071]

[0072] 2. The preparation method of the anti-reagent is as follows:

[0073] (1) Fluorescein isothiocyanate is coupled with luteinizing hormone LH antibody to obtain fluorescein isothiocyanate-labeled luteinizing hormone LH-coated antibody:

[0074] First, use the anti-reagent buffer to prepare the fluorescein isothiocyanate solution with a concentration of 2.5 mg / mL, and the mass ratio of luteinizing hormone LH to fluorescein isothiocyanate is 1:1.3 ...

Embodiment 3

[0086] Embodiment 3: Steps of detecting luteinizing hormone LH with luteinizing hormone LH quantitative assay kit

[0087] The method for detecting luteinizing hormone LH by using this luteinizing hormone LH quantitative assay kit, the method comprises steps:

[0088] (1) Take three test tubes and add 30 μL luteinizing hormone LH calibrator, 30 μL luteinizing hormone LH quality control, and 30 μL sample to be tested;

[0089] (2) Add 60 μL of anti-reagent to each test tube, cover the test tube with a plastic film, shake the test tube gently for 30 s, and place it in a water bath at 37°C for 15 minutes;

[0090] (3) Add 30 μL of magnetic particle reagent to each test tube, cover the test tube with plastic film, shake the test tube gently for 30 seconds, and place it in a water bath at 37°C for 5 minutes;

[0091] (4) Precipitate the test tube on the magnetic separator for 2 minutes, slowly invert the test tube and the magnetic separator, pour out the supernatant, put the inver...

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Abstract

The invention discloses an LH quantitative determination kit. The kit comprises a calibration substance, a quality control substance, a resisting reagent, a magnetic particle reagent and a luminescence substrate. The invention also discloses a making method of the kit, and a method for detecting the LH by using the kit. The resisting reagent is prepared by adopting fluorescein isothiocyanate labeled LH coated antibody and an alkaline phosphatase labeled LH labeled antibody, and the magnetic particle reagent is prepared through coupling an anti-fluorescein isothiocyanate antibody with a carboxyl magnetic sphere, so uniform mixing and separation are easy in an immunization reaction, and the reaction speed is greatly improved; and a novel chemiluminescence substrate APLS is adopted as the substrate, so the sensitivity and the specificity of the kit are improved. The detection kit has the advantages of reliable performances, high sensitivity and wide linear range, and can be used together with a semi-automatic and fully-automatic apparatus.

Description

technical field [0001] The invention belongs to the field of biological immune in vitro diagnosis of medical devices, and mainly relates to a kit for quantitatively detecting luteinizing hormone LH in blood based on magnetic particle separation chemiluminescence method and a preparation method thereof, and the quantitative detection of luteinizing hormone LH in blood by using the kit. Luteinizing hormone LH method. Background technique [0002] Luteinizing Hormone (LH) is a glycoprotein hormone secreted by basophils in the anterior pituitary gland, controlled by hypothalamic gonadotropin-releasing hormone, and is one of the gonadotropins secreted by the pituitary gland. Its main function is to promote Ovulation and corpus luteum production to stimulate the corpus luteum to secrete estrogen (E2) and progesterone (P). The molecular weight of LH is about 28.5kDa. Like other glycoproteins such as FSH, TSH and HCG, it is also composed of α and β subunits. The structures of the ...

Claims

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Application Information

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IPC IPC(8): G01N21/76
Inventor 汪丹杨旻夏振伟
Owner JIANGSU ZECEN BIOTECH CO LTD
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