SU1149940A1 - Method of determining the disturbance of ejaculate fertilizing capability - Google Patents

Abstract

METHOD FOR DEFINITION 0 and the simultaneous increase in the activity of creatine phosphokinase relative to the norm determine the impairment of fertilizing ability Sti e e kulta.

Description

The invention relates to medicine and can be used in the laboratory diagnosis of male infertility and subsequent pathogenetic therapy.

There is a known method for determining the impairment of fertilizing capacity of an eculate by determining the enzymatic activity of eculate ij.

The disadvantage of this method is its low accuracy.

The purpose of the invention is to understand the accuracy of the method.

The goal is to have a kind less than 1.0 and one ™ of a temporary increase in the activity of croquatinophoofokinase relative to the norm # determined k5T impaired fertility -e ejaculate,

The method is carried out as follows.

In the sample of ekulta determine the activity of enzymes: creatine phosphokinase

(CPK), hydroxybutyrate dehydrogenase (HBDG) and lactate dehydrogenase (LDH)

The determination of kreaginphosphokinase gcdroxybutyrate dehydrogenase, n & ktadehydrogenase carried out spectrophotometrically by changing the optical density of the substrate at 340 them for 1 min at 37 ° C for GDDG LDH and at AZS for CPK,

To determine creatine phosphokinase, the reaction mixture contains 20 µl of the test sample of the antiquarian strain prediluted with a physiological solution of 1 g 20 (working solution) and 525 µl of a substrate containing 100 mmol 3 ethanol of a new buffer, pH 7.0, 35 mmol / l creatine phosphate , 20 mmol / l glucose 9 mmol / g glutathione, 10 mmol / l acetic acid magnet 1 mH / l ADP5,. mmol / l NADP, 10 mmol / l AMP, 1.2 kE / l, min, glucose-6-phosphate dehydration, 1.2 kE / l min hexokinase The mixture was stirred and incubated for 5 min.

Then, at 340, they measured the change in optical density in a 1.0 cm cuvette after 1.0 minute, for 3 minutes. The enzyme activity is calculated by the formula; DA / min x 3524 u / l and expressed in international units of e / l.

For the determination of hydroxybutyrate dehydrogenase, the reaction mixture contains 20 µl of a solution of euclet, diluted with 1:20 saline solution, and 630 µl of substrate containing 3sO mmol / l sodium salt (ketobutyrate, 0.18 mmol / l NAD-N and 50 mmol per liter of phosphate buffer, pH 7.5, The resulting mixture is stirred, incubated at 37 ° C for 1.0 mi a {flowed at 340; they measure the change in optical density after 1.0 min in a 1.0 cm cuvette for 3 NIN, the activity of the enzyme is calculated by the formula of 5 dA / min to 5000 U / l and expressed in international units of E / l.

For the determination of lactate dehydrogenase, the reaction mixture contains 20 µl of diluent, diluted with Is 20 with physiological saline, and 630 µl of substrates containing mmol of pyruvate, i-mol / l NADH 8.50 mmol / l of phosphate buffer pH 7 (5) The resulting mixture is incubated at 37 ° C for 1.0 min and the change in optical density in IjO cm is measured for 3 minutes. The activity is calculated according to the formula: Aa / min-x 492 U / l and given in international units E / P,

Calculate the ratio of the magnitude of the activity of HBDG to LDH and, when the activity of CPK is higher relative to the norm (more than 959 U / l) and the ratio of HBDG / LDH is less diagnosed.

Example 1 Freestyle Bee, surveyed for infertile marriage, 25 KST ,, healthful secondary health & YoloBYe signs of vfazheny, Research Edkul ta showed normal sperm content, them. vitality, mobility of 89%, leukocytes isolated, mucus no. The Kurprok-Mshshera test is positive, the Sgogat-Gouner test is positive, and the Basking microagglutination is negative. The data of the proposed method; creatine phosphokinase 1230 U / l HBDG / LDH 0.89 Conclusion: the edema of eculoidum is reduced. The treatment allowed to normalize CPK, HBDG, EDC. Two months after the treatment, the spouse became pregnant.

Example 2. Patient K, examined for infertile marriage, 27 years old, practically healthy. Primary and secondary sexual signs of vfazheny. The study of the prototype eculate revealed normal fertilizing ability. The definition of CK, HBDG, FGD showed that the activity of CK was 949 U / L, and the ratio of HBG to LDH was 1.1, indicating that there was no change in the fertilizing ability of ekult. Recommended comprehensive examination and treatment of the spouse. Later she became pregnant.

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Example 3, Patient P, examined for a barren marriage, 25 years. Practically healthy, Primary and secondary sexual characteristics are high. The study of the prototype eculate revealed a normal fertility ability. The determination of CK, HBDG, LDH showed that the activity of CK was 951 U / L, and the ratio of GDB to DCH was 0.99, which indicated a decrease in the fertilizing ability of ekul. Comprehensive treatment was recommended, after which the spouse became pregnant.

The use of the proposed method in medical practice improves the diagnostic accuracy up to 77.7%. In addition, the method is simple;

performance, can be carried out in laboratory conditions.

Claims (1)

METHOD FOR DETERMINATION OF DISORDERS fertilizing capacity of ejaculate by determining the enzymatic activity of the ejaculate, characterized in that, in order to increase the accuracy of the method, the sample determine the activity of creatine phosphokinase, gydrooksibutiratdegidrogenazy, lactate, calculated ratio gidrooksibutiratdegidrogenazy to lactate dehydrogenase, etc. decrease this ratio below 1 ₃ and simultaneously increasing activity creatine phosphokinases relative to the norm determine a violation of the fertilizing ability of ejacules ta. "1149940 *