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Anti-mullerian hormone chemiluminescence immunoassay kit and preparation method and application thereof

A Mullerian hormone and chemiluminescence immunoassay technology, which is applied in the direction of measuring devices, scientific instruments, instruments, etc., can solve the problems of cumbersome operation process, operation error, inaccurate addition of reagents or samples, etc.

Inactive Publication Date: 2016-10-26
SHENZHEN YHLO BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The traditional AMH detection method mainly uses ELISA, however, limited by the characteristics of ELISA, the traditional AMH detection method is in an open space during the detection process, which is easy to cause cross-contamination between various reagents , and because the detection process is mostly manual, the addition of reagents or samples is not very accurate, the operation process is extremely cumbersome and complicated, prone to operational errors, and the detection accuracy is poor

Method used

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  • Anti-mullerian hormone chemiluminescence immunoassay kit and preparation method and application thereof
  • Anti-mullerian hormone chemiluminescence immunoassay kit and preparation method and application thereof
  • Anti-mullerian hormone chemiluminescence immunoassay kit and preparation method and application thereof

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preparation example Construction

[0063] The preparation process of the anti-Mullerian hormone calibrator is as follows: the calibrator buffer solution is used to prepare anti-Mullerian hormone solutions with concentrations of 0-30 ng / mL respectively.

[0064] Specifically, the anti-Müllerian hormone calibrator is anti-Mullerian hormone with concentrations of 0 ng / mL, 0.7 ng / mL, 2.5 ng / mL, 4.65 ng / mL, 13.58 ng / mL and 28.56 ng / mL Hormone solution.

[0065] The anti-Müllerian hormone chemiluminescence immunoassay kit can use a fully automatic chemiluminescence immunoassay analyzer as a detection tool to complete the detection of the anti-Mullerian hormone. This anti-Müllerian hormone chemiluminescent immunoassay kit, after experiments, has a detection sensitivity of 0.01ng / mL, which is at least 10 times more sensitive than the traditional detection method of anti-Mullerian hormone. The detection accuracy of Mullerian hormone chemiluminescent immunoassay kit is high.

[0066] When this anti-Müllerian hormone ch...

Embodiment 1

[0118] Example 1: Preparation of anti-Müllerian hormone chemiluminescent immunoassay kit

[0119] (1) Preparation of magnetic particles coated with AMH monoclonal antibody:

[0120] Take 50 mg suspension of carboxylated magnetic particles (MagnaBindTM, product number 21353) with a particle size of 0.05 μm to 3 μm, magnetically separate to remove the supernatant, resuspend with 0.02 M MES buffer solution with pH 5.5, add 0.5 mL to 2 mL of new Prepare 10 mg / mL EDC aqueous solution to activate the carboxyl groups on the surface of magnetic beads, add 1 mg ~ 3 mg of AMH monoclonal antibody (Anshlabs, AA011), suspend at room temperature for 2 h ~ 10 h, magnetically separate, remove the supernatant, and wash with 2% BSA 0.1M Tris buffer with a pH of 8.0 was resuspended to 1mg / mL to obtain AMH monoclonal antibody-coated magnetic particles, which were stored in 5mL bottles at 4°C for future use.

[0121] (2) Preparation of acridinium ester-labeled AMH monoclonal antibody:

[0122] T...

Embodiment 2

[0125] Embodiment 2: AMH acridinium ester chemiluminescent immunoassay method

[0126] The present invention uses a fully automatic chemiluminescent immunoassay analyzer and the anti-Müllerian hormone chemiluminescent immunoassay kit prepared in Example 1 as detection tools, and the methodological mode of the present invention is a sandwich method, that is, the instrument adds 50 μL of samples in sequence , 50 μL of AMH monoclonal antibody-coated magnetic particles and 50 μL of acridinium ester-labeled AMH monoclonal antibody. After reacting for 10 minutes, perform magnetic separation. 0.1M HNO 3 and 0.5% H 2 o 2 ) and B solution (containing 0.25M NaOH) for luminescence reaction, and finally record the luminescence intensity, and calculate the AMH content of the tested sample from the standard curve.

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Abstract

The invention discloses an anti-mullerian hormone chemiluminescence immunoassay kit and a preparation method and application thereof. The anti-mullerian hormone chemiluminescence immunoassay kit comprises a solid-phase carrier enveloped by an anti-mullerian hormone monoclonal antibody, and an anti-mullerian hormone monoclonal antibody marked by a chemiluminescence marker. The anti-mullerian hormone chemiluminescence immunoassay kit can complete the assay of anti-mullerian hormone by taking a full-automatic chemiluminescence immunoassay analyzer as an assay tool. Through experiments, the assay sensitivity of the anti-mullerian hormone chemiluminescence immunoassay kit can reach 0.01ng / mL, and is improved by at last ten times compared with the traditional anti-mullerian hormone assay method, and the assay precision of the anti-mullerian hormone chemiluminescence immunoassay kit is higher.

Description

technical field [0001] The invention relates to the field of in vitro detection, in particular to an anti-Müllerian hormone chemiluminescent immunoassay kit and its preparation method and application. Background technique [0002] Müllerian hormone (MIS), also known as Anti-Müllerian hormone (AMH), is a 140kU glycoprotein dimer composed of two 72kDa monomers connected by a disulfide bridge. MIS / AMH belongs to the superfamily of the β-transforming growth factor (TGF-β) family. It was first discovered by Profess Alfred Jost in 1947. AMH is secreted by the immature Sertoli cells of the testis and the granulosa cells of the growing follicles of the postnatal ovary. In males, it is initially expressed in the Sertoli cells of the 8-week-old fetus, beginning at embryogenesis and continuing throughout life. During embryonic development, AMH causes the degeneration of the Mullerian ducts, which form the normal male reproductive tract. For women, AMH is produced by follicular gran...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/577
CPCG01N33/54326G01N33/577
Inventor 夏福臻钱纯亘王刚祝亮刘美婷
Owner SHENZHEN YHLO BIOTECH
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