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Human anti-mullerian hormone determination kit and preparation method and application thereof

A detection kit and kit technology, applied in the field of homogeneous luminescent immunoassay kits for anti-human Mullerian hormone, can solve the problems of poor repeatability of enzyme immunoassays, prone to "hook effect, environmental pollution, etc., and achieve Good amplification effect, high accuracy and good sensitivity

Active Publication Date: 2018-08-24
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The problem to be solved by the present invention is to provide a serum anti-Mullerian hormone homogeneous luminescent immunoassay kit and its preparation and use method, so as to solve the cumbersome washing steps of the existing heterogeneous immunoassay kits. At the same time, it overcomes the defects of environmental pollution and short shelf life of radioimmunoassay, and overcomes the defect of poor repeatability and prone to "hook effect" of enzyme immunoassay

Method used

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  • Human anti-mullerian hormone determination kit and preparation method and application thereof
  • Human anti-mullerian hormone determination kit and preparation method and application thereof
  • Human anti-mullerian hormone determination kit and preparation method and application thereof

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preparation example Construction

[0089] The first aspect of the present invention provides a method for preparing a human anti-Müllerian stimulation homogeneous detection kit, the method comprising:

[0090] preparing a first component, the first component comprising a receptor and a first binding unit bound thereto, the receptor capable of reacting with singlet oxygen to generate a detectable signal, the first binding unit capable of binding a human antibody The first epitope of Müllerian activation;

[0091] preparing a second component, said second component comprising a first label and a second binding unit bound thereto, said second binding unit being capable of binding a second epitope of human anti-Müllerian hormone, said The second epitope and the first epitope are human anti-Müllerian epitopes with different binding properties or epitopes with the same binding properties at different positions;

[0092] A third component is prepared comprising a donor capable of generating singlet oxygen in an excit...

Embodiment 1

[0169] Embodiment 1-basic kit

[0170] The core components of the kit are:

[0171] The AMH quantitative assay detection kit (light-induced chemiluminescence method) of the present embodiment consists of a composition (reagent 1, R1) containing luminescent microspheres (FG-Ab1) coated with the first anti-AMH monoclonal antibody, biotin The composition (reagent 2, R2) of the labeled second anti-AMH monoclonal antibody (Bio-Ab2); also includes the sample to be tested. In addition, it includes a composition (reagent 3, R3) comprising photosensitive microspheres (SA-GG) coated with streptavidin, which is prepared according to the method of the aforementioned materials and equipment, including light-induced chemiluminescence analysis instrument and so on.

[0172] The using method of the test kit of the present embodiment comprises:

[0173] The detection process is fully automatically completed by the automatic light-induced chemiluminescence analysis system and the detection r...

Embodiment 2

[0182] Embodiment 2-core kit

[0183] Experiment of the present invention:

[0184] Preparation of kit materials:

[0185] The preparation methods of specific materials and corresponding reagents are prepared according to the materials and equipment section.

[0186] The using method of the test kit of the present embodiment comprises:

[0187] Reagents are added in parallel in different reaction wells, parallel detection is performed, the reaction wells are irradiated with excitation light with a wavelength of 680nm, and emission light with a wavelength of 612nm is detected.

[0188] The specific detection steps are:

[0189] a. Add 25 μl samples to different reaction wells; the concentrations of AMH in the calibrator samples are 0, 0.16, 0.6, 4, 10, and 24 ng / mL respectively. For the concentrations of AMH in the samples to be tested, refer to the lower half of Table 1.

[0190] b. Add 25 μl R1 and 25 μl R2 in sequence to each reaction well;

[0191] c. Incubate at 37°C ...

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Abstract

The invention discloses a preparation method of a human anti-mullerian hormone AMH homogeneous detection kit, and the method is as follows: a component comprising a receptor and a first binding unit bound to the receptor is prepared, the receptor can be reacted with singlet oxygen to generate a detectable signal, the first binding unit is capable of binding to the first epitope of anti-mullerian hormone AMH; a second component comprising a first label and a second binding unit bound to the first label is prepared, the second binding unit is capable of binding to the second epitope of the AMH,the first epitope and the second epitope of the AMH are epitopes with different binding properties or epitopes with a same binding property at different positions; and a third component comprising a donor and a first label specific binder bound to the donor is prepared, and the donor is capable of generating the singlet oxygen in an excited state. The kit prepared by the method can efficiently detect the AMH.

Description

technical field [0001] The invention belongs to the technical field of biomolecular detection, and relates to a homogeneous immunoassay kit without matrix effect, in particular to an anti-human Mullerian hormone homogeneous luminescent immunoassay kit and its preparation and use methods. Background technique [0002] Anti-Mullerian hormone (AMH) is a dimer glycoprotein belonging to the transforming growth factor-β family, as an inhibitor of Mullerian tubules. During the development of gonad organs, AMH plays an important role and is one of the important substances to ensure the development of male and female organs and the function of gonads. In males, AMH is mainly produced by Leydig cells of the testis, starting from embryogenesis and throughout life; during male fetal development, AMH leads to the degeneration of Mullerian ducts and the formation of male reproductive tracts. The AMH content of males is higher in the neonatal period, and it still rises a few months after ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/543G01N33/533
CPCG01N33/533G01N33/54313G01N33/577
Inventor 徐静心史舟芳强中华李会强刘宇卉李临
Owner CHEMCLIN DIAGNOSTICS CO LTD
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