Kit for detecting activity of acid phosphatase in seminal plasma and detection method

A technology of acid phosphatase and active reagents, which is applied in biochemical equipment and methods, microbiological determination/inspection, color/spectral characteristic measurement, etc. It can solve the problems of many operation steps, difficult to widely use, large amount of reagents, etc., to achieve Easy to operate, widely applicable, and the effect of saving the amount of reagents

Inactive Publication Date: 2011-02-16
南京欣迪生物药业工程有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has many steps, requires a large amount of reagents, and requires a standard curve, making it difficult to be widely used in clinical routine testing.

Method used

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  • Kit for detecting activity of acid phosphatase in seminal plasma and detection method
  • Kit for detecting activity of acid phosphatase in seminal plasma and detection method
  • Kit for detecting activity of acid phosphatase in seminal plasma and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: A kit for detecting seminal plasma acid phosphatase activity of the present invention includes a matrix fluid, a standard product, a stop fluid and a color developing agent. The matrix solution is a phenyl disodium phosphate matrix solution with a concentration of 0.01 mol / L; the standard is a phenol standard solution with a concentration of 1 mg / ml; the stop solution is an alkaline solution, in 1 L of the alkaline solution Contains 21g sodium bicarbonate, 0.5g 4-aminoantipyrine and 10g sodium hydroxide. The color developing agent is a potassium ferricyanide solution, and 1 L of the potassium ferricyanide solution contains 2.5 g of potassium ferricyanide and 17 g of boric acid. Calculated according to 100 test reagents required for each kit loaded, the matrix solution is 50ml±2ml, the phenol standard solution is 0.4ml±16μl, the stop solution is 20ml±0.8ml, and the color reagent is 30ml±1.2ml.

Embodiment 2

[0021] Example 2: A method for detecting seminal plasma acid phosphatase activity using a seminal plasma acid phosphatase activity detection kit in Example 1. The specific steps of the method are as follows:

[0022] 1) Setting the parameters of the detection instrument: the type of the detection instrument: end point method; dominant wavelength: 510nm; delay time: 3s; measurement time: 3s; project unit: U / ml; standard concentration: 33.3 U / ml; cuvette Temperature: 37℃, the detection instrument is a semi-automatic biochemical analyzer;

[0023] 2) Add phenyl disodium phosphate matrix solution to the blank tube, add the standard product and phenyl disodium phosphate matrix solution to the standard tube, add diluted seminal plasma and phenyl disodium phosphate matrix solution to the sample tube;

[0024] 3) Incubate the above blank tube, standard tube and sample tube at the same time, with an incubation time of 15 minutes and an incubation temperature of 37°C;

[0025] 4) Then add the s...

Embodiment 3

[0026] Example 3: Determine the volume of seminal plasma suitable for the reaction system according to the detection method of Example 2

[0027] The seminal plasma sample is diluted 1:1000 with saline, and then take 1, 2, 3, 4, 5μl of the diluted seminal plasma, add 500μl matrix solution, mix and incubate at 37℃ for 15min, add 200μl alkaline solution and 300μl potassium ferricyanide solution, after mixing, check the optical density value of each tube. The amount of diluted seminal plasma with an optical density value between 0.2 and 0.4 is the best. The results show that a 4μl seminal plasma reaction system is more suitable.

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Abstract

The invention discloses a kit for detecting the activity of acid phosphatase in seminal plasma and a detection method. The kit comprises substrate solution, a standard sample, stop solution and a color development reagent, wherein the substrate solution is disodium phenyl phosphate substrate solution; and the stop solution is alkaline solution. The detection method comprises the following steps of: first setting parameters of a detecting instrument; then adding the disodium phenyl phosphate substrate solution into an empty tube, adding the standard sample and the substrate solution into a standard tube, and adding the dilute seminal plasma and the substrate solution into a sample tube; next simultaneously performing incubation on the empty tube, the standard tube and the sample tube at the temperature of 37 DEG C for 15 minutes; and finally adding the same amount of alkaline solution and color development reagent into the incubated empty tube, the incubated standard tube and the incubated sample tube, performing uniform mixing, and performing color comparison by using the detecting instrument to obtain a result. The kit and the detection method of the invention can make the operation simple, convenient, time-saving and reagent-saving, and can make stable and reliable the result and be applied to laboratories for the department of andrology in various hospitals as synchronous comparison tests can be performed.

Description

Technical field [0001] The invention relates to a biochemical kit and a detection method, in particular to a kit and a detection method for detecting seminal plasma acid phosphatase activity. Background technique [0002] About 30% of human seminal plasma comes from the prostate, and the function of the prostate is of great significance to male fertility. Seminal plasma acid phosphatase (ACP) activity detection is considered to be one of the important indicators for evaluating male prostate function. ACP activity often decreases during prostatitis, and ACP can also be used as an indicator for the diagnosis and treatment of prostate cancer. Seminal plasma ACP has immunosuppressive effects and is an important component of seminal immunosuppressive agents. In addition, ACP can also be used as an experimental indicator of abnormal semen liquefaction, and has a wide range of applications in the judgment of forensic rape cases. [0003] At present, some andrology laboratories in my cou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/42G01N21/31
Inventor 陆金春陈晓慧卢坤刚
Owner 南京欣迪生物药业工程有限责任公司
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