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Kit for detecting seminal gamma-L-glutamyltranspetidase and detection method

A technology of glutamyl transpeptidase and kit, which is applied in the fields of biochemical equipment and methods, measurement/inspection of microorganisms, measurement of color/spectral characteristics, etc., and can solve problems that are difficult to be widely used, many operation steps, expensive reagents, etc. problems, achieving stable and reliable results, easy operation, and saving reagent consumption

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

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Problems solved by technology

This method has many steps, time-consuming and reagent-consuming, and the seminal plasma needs to be diluted before detection, so it is difficult to be widely used in clinical routine detection

Method used

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  • Kit for detecting seminal gamma-L-glutamyltranspetidase and detection method
  • Kit for detecting seminal gamma-L-glutamyltranspetidase and detection method
  • Kit for detecting seminal gamma-L-glutamyltranspetidase and detection method

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Effect test

Embodiment 1

[0017] Embodiment 1: A kit for detecting seminal plasma γ-GT of the present invention, including α-naphthylamine standard solution, γ-glutamyl-α-naphthylamine matrix solution and diazo reagent. Wherein the α-naphthylamine standard solution has a concentration of 2.86mg / ml; the γ-glutamyl-α-naphthylamine matrix solution has a concentration of 2.71mg / ml; the diazo reagent consists of 1g / L sodium nitrite aqueous solution and 2g / L The L aminobenzenesulfonic acid solution can be mixed at a volume ratio of 1:4, wherein every 100ml aminobenzenesulfonic acid solution contains 20ml of acetic acid. Calculated according to the detection reagents required for loading 100 tests in each kit, the α-naphthylamine standard solution is 0.1ml±4μl, the γ-glutamyl-α-naphthylamine matrix solution is 30ml±1.2ml, and the diazo reagent is 200ml±8ml.

Embodiment 2

[0018] Embodiment 2: A method for detecting seminal plasma γ-GT with a test kit for detecting seminal plasma γ-GT in Example 1, the specific steps of the method are as follows:

[0019] 1) Setting detection instrument parameters: the detection instrument is a semi-automatic biochemical analyzer, and its parameters are: the item type is an endpoint method; the dominant wavelength is 510nm; the delay time is 3s; the measurement time is 3s; the item unit is U / ml ; Standard concentration is 80U / ml; Cuvette temperature is 37°C;

[0020] 2) Add matrix solution to the blank tube, add standard solution and matrix solution to the standard tube, and add seminal plasma and matrix solution to the sample tube;

[0021] 3) Incubate the above-mentioned blank tube, standard tube and sample tube at the same time, the incubation time is 15min, and the incubation temperature is 37°C; the seminal plasma is the supernatant obtained by centrifuging the liquefied semen at 3000g for 15min;

[0022] ...

Embodiment 3

[0023] Embodiment 3: according to the detection method of embodiment 2, the reaction volume suitable for the reaction system is determined

[0024] According to the results of manual detection, one case of seminal plasma sample with γ-GT activity near the upper limit of the normal reference range was selected. Take 100, 200, 300, and 400 μl of matrix solution, make up each tube with normal saline to a total volume of 400 μl, add 1 μl of seminal plasma sample, mix well and incubate at 37°C for 15 minutes, add 5ml of diazonium reagent, and incubate again at 37°C 5min, 510nm to detect the absorbance of each tube. According to the principle that the amount of matrix should be excessive and the reaction can be fully carried out, it is found that 300 μl of matrix solution and 1 μl of seminal plasma are more suitable for the reaction. On this basis, take 4 test tubes, add 300 μl matrix solution and 1 μl seminal plasma sample respectively, mix well and incubate at 37°C for 15 minutes...

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Abstract

The invention discloses a kit for detecting seminal gamma-L-glutamyltranspetidase and a detection method. The kit for detecting the seminal gamma-L-glutamyltranspetidase comprises alpha naphthylamine standard liquid, gamma-glutamoyl-alpha-naphthylamine matrix liquid and diazonium reagent. The detection method comprises the following steps of: setting parameters of a detection instrument; adding the matrix liquid into a blank tube, adding the standard liquid and the matrix liquid into a standard tube, and adding semen and the matrix liquid into a sample tube; simultaneously incubating the blank tube, the standard tube and the sample tube for 15 minutes at the temperature of 37 DEG C; and adding the diazonium reagent, mixing the diazonium reagent and the liquid uniformly, then incubating the tubes again for 5 minutes at 37 DEG C, and comparing colors through the detection instrument to obtain results. The kit and the detection method can simplify the operation, save the time and save the reagent; and the synchronous comparison experiments can be performed so that the results are stable and reliable, and the kit and the detection method are suitable for male laboratories of various hospitals.

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 γ-L-glutamyl transpeptidase. Background technique [0002] Seminal plasma is rich in γ-L-glutamyl transpeptidase (γ-GT), which is a key enzyme in glutathione (GSH) metabolism and plays an important role in protecting sperm from oxygen free radical damage. The γ-GT in semen mainly comes from the prostate, and the detection of its activity can be used to evaluate the function of the prostate. γ-GT in seminal plasma can decompose γ-glutamyl-α-naphthol into free α-naphthol, and α-naphthol reacts with diazo reagent to produce red color, which is proportional to the enzyme activity. [0003] At present, there are very few andrology laboratories that carry out semi-automatic γ-GT activity detection in seminal plasma. On the basis of the manual detection of semi-automatic γ-GT in semi-automatic plasma, we have further e...

Claims

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Application Information

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IPC IPC(8): C12Q1/48G01N21/31
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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