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Seminal fluid leukocyte group flow type quantitative detection reagent, kit and detection method

A detection reagent and quantitative detection technology, which is applied in the field of seminal leukocyte flow cytometry quantitative detection reagents, can solve the problems of non-specific operation, strong subjective factors, cumbersome and other problems, and achieve the effect of specificity, convenience, easy access, and easy operation

Pending Publication Date: 2021-10-19
THE THIRD PEOPLES HOSPITAL OF SHENZHEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In view of the above-mentioned shortcomings of the prior art, the object of the present invention is to provide a flow cytometry quantitative detection reagent, kit and detection method for semen leukocyte populations, which are used to solve the subjective factors existing in the quantitative detection methods of semen leukocyte populations in the prior art Strong, non-specific and cumbersome to operate

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  • Seminal fluid leukocyte group flow type quantitative detection reagent, kit and detection method

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Experimental program
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Embodiment 1

[0132] A flow cytometric quantitative detection reagent for semen leukocyte populations, comprising:

[0133] 1) Sample treatment solution: containing Na 2 HPO 4 、KH 2 PO 4 , NaCl solution, Na 2 HPO 4 、KH 2 PO 4 , NaCl concentrations are 1.42g / L, 0.27g / L, 8g / L respectively;

[0134] 2) Sample diluent: containing Na 2 HPO 4 、KH 2 PO 4 , NaCl and KCl solutions, Na 2 HPO 4 、KH 2 PO 4 , NaCl and KCl concentrations are 1.42g / L, 0.27g / L, 8g / L, 0.2g / L respectively;

[0135] 3) Sample separation liquid: a solution containing polysucrose and diatrizoate, by mass percentage, including: 5% diatrizoate, 4.3% polysucrose, 1.7% polyethylene glycol, and the balance is pyrogen-free Ionized water;

[0136] 4) Antibody solution: select the IgG-type monoclonal antibody with fluorescent labeling anti-human CD45 derived from rabbits as the antibody, and dissolve fetal bovine serum FBS and the antibody in deionized water to prepare the antibody solution. The FBS concentration is 0....

Embodiment 2

[0140] A flow cytometric quantitative detection reagent for semen leukocyte populations, comprising:

[0141] 1) Sample treatment solution: containing Na 2 HPO 4 、KH 2 PO 4 , NaCl solution, Na 2 HPO 4 、KH 2 PO 4 , NaCl concentrations are 1.30g / L, 0.19g / L, 7g / L respectively;

[0142] 2) Sample diluent: containing Na 2 HPO 4 、KH 2 PO 4 , NaCl and KCl solutions, Na 2 HPO 4 、KH 2 PO 4 , NaCl and KCl concentrations are 1.30g / L, 0.19g / L, 7g / L, 0.1g / L, respectively;

[0143] 3) Sample separation liquid: a solution containing polysucrose and diatrizoate, by mass percentage, including: 4% diatrizoate, 5.0% polysucrose, 1.0% polyvinylpyrrolidone, and the balance is deionized without pyrogen water;

[0144] 4) Antibody: select the IgG type monoclonal antibody with fluorescence labeling anti-human CD45 derived from rabbit as antibody, dissolve fetal bovine serum FBS and antibody with deionized water to prepare antibody solution, FBS concentration is 0.01%, antibody concen...

Embodiment 3

[0148] A flow cytometric quantitative detection reagent for semen leukocyte populations, comprising:

[0149] 1) Sample treatment solution: containing Na 2 HPO 4 、KH 2 PO 4 , NaCl solution, Na 2 HPO4 、KH 2 PO 4 , NaCl concentrations are 1.50g / L, 0.39g / L, 9g / L respectively;

[0150] 2) Sample diluent: containing Na 2 HPO 4 、KH 2 PO 4 , NaCl and KCl solutions, Na 2 HPO 4 、KH 2 PO 4 , NaCl and KCl concentrations are 1.50g / L, 0.39g / L, 9g / L, 0.3g / L, respectively;

[0151] 3) Sample separation liquid: a solution containing polysucrose and diatrizoate, by mass percentage, including: 4% diatrizoate, 4.0% polysucrose, 2.0% polystyrenesulfonic acid, and the balance is pyrogen-free Deionized water;

[0152] 4) Antibody: select the IgG type monoclonal antibody with fluorescence labeling anti-human CD45 derived from rabbit as antibody, dissolve fetal bovine serum FBS and antibody with deionized water to prepare antibody solution, the concentration of FBS is 0.01%, and the c...

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Abstract

The invention discloses a seminal fluid leukocyte group flow type quantitative detection reagent, a kit and a detection method. The detection reagent and the kit comprise: (1) a sample treating agent containing Na2HPO4, KH2PO4 and NaCl; (2) a sample diluent, wherein the sample diluent contains Na2HPO4, KH2PO4, NaCl and KCl; (3) a sample separating agent containing polysucrose and meglumine diatrizoate; (4) an antibody, namely an IgG type monoclonal antibody of anti-human CD45 with a fluorescence label; and (5) a sample fixing agent containing paraformaldehyde. The detection method is based on a flow cytometry principle and comprises the following steps: removing impurities and non-target cells in the seminal fluid by using a sample treating agent, specifically binding a leukocyte CD45 antibody to the surfaces of leukocytes in the seminal fluid, and finally detecting specific marker fluorescence by using a flow cytometry, thereby obtaining the number of leukocytes. The method is detected and recorded by a machine, is more objective and stable than a peroxidase method, and is easy to standardize and high in specificity; and an antibody for dyeing is stable in property, long in storage time and convenient to take and use.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnostic reagents, kits and detection methods, in particular to a quantitative detection reagent, kit and detection method for semen white blood cell population flow cytometry. Background technique [0002] In recent years, due to the improvement of productivity and the use of a large number of mechanized and automated equipment, men have gradually become the main force of sedentary, and reproductive tract infection caused by sedentary has become an important cause of male infertility. The male infertility rate is increasing year by year, and the specificity and accuracy of the diagnosis of male reproductive tract infection are getting higher and higher, and the voice is getting stronger and stronger. Leukocytospermia is an important cause of male infertility, which means that the concentration of white blood cells in semen exceeds 1×10 6 / ml, suggesting that there may be infection in the repr...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N15/14
CPCG01N33/56972G01N15/1434G01N2015/1006G01N2015/016
Inventor 曲久鑫
Owner THE THIRD PEOPLES HOSPITAL OF SHENZHEN
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