Luteinizing hormone nano-magnetic particle chemiluminescence quantitative immunoassay kit and preparation method thereof

A luteinizing hormone and chemiluminescence immunological technology, applied in the field of immunoassay medicine, can solve the problems of short shelf life, narrow detection range and high cost, and achieve the effects of good stability, good specificity and good performance.

Active Publication Date: 2013-09-11
THE SECOND AFFILIATED HOSPITAL OF NANJING MEDICAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The problem to be solved by the present invention is to provide a chemiluminescent immunoassay quantitative detection kit for luteinizing hormone and its preparation method, which avoids the shortcomings of radioimmunoassay reagents su

Method used

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  • Luteinizing hormone nano-magnetic particle chemiluminescence quantitative immunoassay kit and preparation method thereof
  • Luteinizing hormone nano-magnetic particle chemiluminescence quantitative immunoassay kit and preparation method thereof

Examples

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Effect test

Embodiment 1

[0054] Example 1: Preparation of Luteinizing Hormone Nano-Magnetic Particle Chemiluminescent Immunoquantitative Assay Kit

[0055] (1) Preparation of luteinizing hormone calibrator:

[0056] The luteinizing hormone antigen (manufactured by Fitzgerald Company) was prepared with goat serum (purchased from Zhengzhou Yikang Bioengineering Co., Ltd.) to make a calibrator concentrated stock solution, and the national calibrator (batch number: 150531-0211, specification: 530mIU / bottle) For calibration, dilute the concentrated stock solution with goat serum to the working concentration, respectively 0, 2, 10, 25, 100, 250mIU / mL, which is the concentration of the calibrator;

[0057] (2) Preparation of luteinizing hormone quality control products:

[0058] Dilute the above concentrated stock solution to 5mIU / mL and 120mIU / mL with goat serum respectively, and calibrate with the national calibrator; 5mIU / mL is used as a low-value quality control product, and 120mIU / mL is used as a high-...

Embodiment 2

[0076] Embodiment 2: the inspection of kit of the present invention

[0077] (1) Physical inspection: liquid components should be clear without sediment or floc; other components should have no package damage.

[0078] (2) Accuracy: The kit calibration product and the national standard product series are analyzed and determined at the same time, and the double-logarithmic mathematical model is used for fitting, requiring that the two dose-response curves do not deviate significantly from parallel (t test, |t|<2.447) ; With the national standard substance of luteinizing hormone as the reference substance, the average value of the ratio between the measured value and the labeled value of the kit calibrator should be within the range of 0.90 to 1.10 by using a double-logarithmic mathematical model.

[0079] (3) Linearity of the dose-response curve: Fitting with a double-reading mathematical model, the absolute value of the correlation coefficient r of the dose-response curve with...

Embodiment 3

[0087] Embodiment 3: the using method of kit of the present invention

[0088] (1) Equilibrate the test kit at room temperature (18-25°C) for 30 minutes.

[0089] (2) Preparation of lotion: dilute the concentrated lotion with distilled water at 1:20 (1mL of lotion plus 19mL of distilled water). If the concentrated lotion has crystals, the concentrated lotion can be placed at room temperature or 37°C, and then diluted after the crystals dissolve.

[0090] (3) Preparation of luminous liquid: Take an appropriate amount of luminous liquid A and luminous liquid B and mix them in equal volumes 5 minutes before use.

[0091] (4) Number the reaction tubes, add 25-50uL calibrator or serum samples, 50uL magnetic particle-streptavidin suspension, 50uL biotin-luteinizing hormone antibody conjugate, 50-100uL Luteinizing hormone antibody enzyme conjugate, shake reaction at 37°C for 30 minutes, place the test tube rack on a magnetic separator for separation for 5 minutes, then pour out the...

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Abstract

The invention discloses a luteinizing hormone nano-magnetic particle chemiluminescence quantitative immunoassay kit. The kit comprises a luteinizing hormone calibration product, a nano-magnetic particle suspension coupled with streptavidin, a biotin-marked luteinizing hormone antibody, a luteinizing hormone antibody enzyme binding substance, a luteinizing hormone quality control product, a chemiluminescence solution A and a chemiluminescence solution B, a 20-times concentrated washing solution and a reaction tube, wherein the used enzyme is horseradish peroxidase, the purity RZ of the horseradish peroxidase is not less than 3.0, and the activity is not less than 250U/mL. In addition, the invention further discloses a preparation method of the kit. Compared with the existing kit, the kit disclosed by the invention has the advantages of high sensitivity, a wide range of measurable concentrations, long effective period of a reagent, high degree of automation in detection and the like, and is simple to operate.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine. Specifically, the invention provides a luteinizing hormone (LH) nano magnetic particle chemiluminescent immunoquantitative detection kit and a preparation method thereof. Background technique [0002] Luteinizing hormone (Luteotropic hormone, LH) is secreted by adenohypophysis basophils. In men, it can stimulate the Leydig cells of the testis to secrete male hormones, and in women, it can stimulate the ovaries to secrete female estrogen. Luteinizing hormone (LH) cooperates with follicle stimulating hormone (FSH) in women to maintain the menstrual cycle of the ovaries, leading to ovulation and corpus luteum formation. The production of LH is controlled by the hypothalamic gonadotropin-releasing hormone, and is also regulated by positive and negative feedback from the ovary. The combined detection of LH and FSH is mainly used to identify primary (ovarian) or secondary (pituitary) amenorrhea...

Claims

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Application Information

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IPC IPC(8): G01N33/74G01N33/531G01N21/76
Inventor 苏东明刘萍梁秀彬栾大伟郭万华刘云
Owner THE SECOND AFFILIATED HOSPITAL OF NANJING MEDICAL UNIV
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