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Reagent for detecting sperm membrane surface antigen by immuno-magnetic sand method, its preparation and detection method

A technology for detecting reagents and immunobeads, which is applied in the field of sperm membrane surface antibody immunobead method detection reagents and its preparation and detection. It can solve the problems of different sizes of immune beads, difficult to obtain, and small males, and overcome the inability to directly detect , The method is simple and easy to implement, and the effect is easy to interpret

Inactive Publication Date: 2006-04-05
深圳华康生物医学工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, due to the difficulty in obtaining the detection reagents of the immunobead method, enzyme-linked immunoassay (ELISA) is mainly used in China to detect anti-sperm antibodies in serum or seminal plasma. This test is of little significance to men, and the false positive rate is high.
In the actual operation of the immune bead test, there are also differences in the size of the immune beads, and the test results are difficult to interpret, such as figure 1 shown
At the same time, there are shortcomings such as the inability to directly detect samples of asthenospermia and oligospermia

Method used

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  • Reagent for detecting sperm membrane surface antigen by immuno-magnetic sand method, its preparation and detection method
  • Reagent for detecting sperm membrane surface antigen by immuno-magnetic sand method, its preparation and detection method

Examples

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Embodiment 1

[0034] Embodiment 1, a kind of sperm membrane surface antibody immunobead method detection reagent, comprises

[0035] 1) The immune beads stored in the diluent: the main components of the diluent are pH 7.4, containing Nacl125mmol / L, bovine serum albumin 1.5g / L, NaN 3 0.15%, 0.02mol / L phosphate buffer solution of phosphate solute, said immune beads are carboxylated polystyrene latex particles with a diameter of about 2.0 μm cross-linked with anti-human IgG or IgA or IgM antibodies;

[0036] 2) Washing solution: Tyrode's solution containing 0.3% bovine serum albumin;

[0037] 3) Suspension: Tyrode's solution containing 5% bovine serum albumin;

[0038] 4) High motility sperm extract: containing NaCl 0.7%, KCl 0.045%, CaCl 2 2H 2 O 0.025%, MgSO 4 ·7H 2 O 0.015%, NaHCO 3 0.25%, NaH 2 PO 4 2H 2 O 0.015%, sodium pyruvate 0.0025%, 60% sodium lactate as syrup 0.35%, NaN 3 0.015%, glucose 0.13%, citric acid monohydrate 0.035%, calf serum 0.09% aqueous solution;

[0039] Am...

Embodiment 2

[0047] Embodiment two, a method for preparing the sperm membrane surface antibody immunobead method detection reagent in embodiment one, comprising the following steps,

[0048] 1) Preparation and dilution of immune beads:

[0049] a) Preparation of immunobead dilution: in the phosphate buffer solution with a concentration of 0.02mol / L, add 125mmol / L of Nacl, 1.5g / L of bovine serum albumin and 0.15% NaN 3 , adjusted the pH to 7.4, filtered through a 0.22 μm filter and stored.

[0050] b) Preparation of immune beads: In 2.5ml of carbonate buffer solution with a concentration of 0.5mol / L, add 0.3% acrylic resin S-100, adjust the pH to 9.5, and carboxylated polyphenylene with a particle diameter of about 2.0 μm Ethylene latex was mixed and placed at 5° C. for 1 hour to fully react.

[0051] The carboxylated polystyrene latex with a particle diameter of about 2.0 μm is obtained from carboxylated polystyrene latex with a concentration of 2.5% produced by Sigma Company of the Unit...

Embodiment 3

[0063] Embodiment three, a method for detecting sperm membrane surface antibody using the sperm membrane surface antibody immunobead method detection reagent prepared in embodiment two, comprising the following steps,

[0064] 1) Reagent preparation

[0065] Washing of the immune beads: the immune beads can be washed before use to avoid non-specific adhesion of the immune beads to the surface of the sperm. The specific method is as follows:

[0066] a) Add 50 μl of immune beads diluted to the optimum concentration and 1.5 ml of washing solution to the Eppendorf tube, and centrifuge at 2000 g for 10 min;

[0067] b) Discard the supernatant and add the resuspension solution to 50 μl.

[0068] Store the washed immunobeads at 2-8°C for 1 month.

[0069] 2) Sample preparation

[0070] a) Ordinary semen sample preparation

[0071] ① Transfer a certain volume of fresh semen to a conical bottom centrifuge tube, and add washing solution to 4ml. The amount of semen sample can refer...

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Abstract

The invention discloses a semen film surface antibody immunity method testing agent and preparing and testing method. The semen film surface antibody immunity method testing agent comprises: 1.8um-2.2um immunity bead which can possesses human antibody IgG or IgA or IgM micromole rubber latex particle, the washing liquid which contents 0.1%-0.5% bonnie serum albumin and the scale dissolver which contents 4%-6% bonnie serum albumin. The agent can also comprise semen extracting liquid with high activity. The invention provides the method for preparing the semen film surface antibody immunity method testing agent.

Description

【Technical field】 [0001] The invention relates to a reagent for in vitro diagnosis and its preparation and detection method, in particular to a sperm membrane surface antibody immune bead method detection reagent and its preparation and detection method. 【Background technique】 [0002] Immune factors are one of the causes of male infertility, so its clinical detection is a very important topic. In 2000, the World Health Organization (WHO) stipulated that mixed agglutination reaction (MAR) and immunobead test (IBT) were standard tests for diagnosing immune infertility. [0003] At present, due to the difficulty in obtaining the detection reagents of the immunobead method, enzyme-linked immunoassay (ELISA) is mainly used in China to detect anti-sperm antibodies in serum or seminal plasma. This test is of little significance to men, and the false positive rate is high. . In the actual operation of the immune bead test, there are also differences in the size of the immune bead...

Claims

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Application Information

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IPC IPC(8): G01N33/546G01N33/531
Inventor 傅剑华刘瑜胡家纯何林何小红
Owner 深圳华康生物医学工程有限公司
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