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
View PDF7 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This technique has low sensitivity, narrow linearity, poor specif

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • 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

Example Embodiment

[0053] Example 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 1L 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 it, and process it with 0.22μm filter membrane.

[0059] 3. Anti-reagent buffer

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

[0061] 4. Magnetic particle buffer

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

[0063] 5. Luminescent substrate buffer

[0064] Tris 12.12g~121.14g, sodium chloride 5.82g, lucigenin 0.03g, add 1L purified water, stir we...

Example Embodiment

[0067] Example 2: Preparation of kit for quantitative determination of luteinizing hormone LH

[0068] 1. Preparation of calibrators and quality control products

[0069] First: Dissolve the luteinizing hormone LH with a standard buffer solution, and configure it as a calibrator and quality control product of the target concentration shown in Table 1. The luteinizing hormone LH used in this example is purchased from a well-known domestic and foreign manufacturer Fitzgerald .

[0070] Table 1: Preparation of calibrators and quality control products

[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 luteinizing hormone LH coated antibody labeled with fluorescein isothiocyanate:

[0074] First, use anti-reagent buffer to prepare fluorescein isothiocyanate with a concentration of 2.5 mg / mL fluorescein isothiocyanate solution, according to the mass ratio of luteinizin...

Example Embodiment

[0086] Example 3: Steps for detecting luteinizing hormone LH with a kit for quantitative determination of luteinizing hormone LH

[0087] The method for detecting luteinizing hormone LH by using this luteinizing hormone LH quantitative determination kit includes the steps:

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

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

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

[0091] (4) Settling 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 inve...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G01N21/76
Inventor 汪丹杨旻夏振伟
Owner JIANGSU ZECEN BIOTECH CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap