SU1640653A1 - Method for diagnosis of disseminated sclerosis - Google Patents

Abstract

The invention relates to medicine, in particular to methods for diagnosing multiple sclerosis. The aim of the invention is to improve accuracy. A method for diagnosing multiple sclerosis is performed by taking blood from a patient before and 30 minutes after administering a solution of reserpine and determining the number of immune lymphocyte sockets of blood with sheep erythrocytes loaded with serotonin in both samples. At a constant value of the indicator compared with the initial one, multiple sclerosis is diagnosed. The accuracy of diagnosis increases by 17% to 22%.

Description

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The invention relates to the field of medicine, namely to the laboratory diagnosis of diseases, which can be used to recognize multiple sclerosis (PC).

The purpose of the invention is to improve the accuracy of diagnosis of multiple sclerosis.

The method is carried out as follows.

0.05-0.1 ml of blood is taken from the finger of a patient with PC in 0.05-0.1 ml of 5% sodium citrate solution to prevent blood from clotting. 0.5 ml of a 2.5% reserpine solution is injected subcutaneously into the patient. After 30 minutes, blood is re-taken as previously described. After thoroughly shaking the tube, 0.8 to 0.9 ml of distilled water is added to the erythrocyte dosage for 30 seconds, and then it is filled up to 9-10 ml

medium 199 for recovery of pH 7.0. The tubes are centrifuged for 5-10 minutes at 1500 rpm, then the supernatant is drained and the leukocyte pellet is resuspended in 0.1-0.2 ml of isotonic sodium chloride solution or medium 199. To the resulting suspension add 0.1-0. 2 ml of a 0.5% suspension of glutaurized erythrocytes of a ram loaded with serotonin, the mixture is incubated for 10-15 minutes at + 37.0-37.5 ° C, then centrifuged at 1500 rpm for 5-10 minutes. The supernatant is removed with a pipette, the precipitate is resuspended in a drop of any intact serum (bovine, horse, human). A smear is made from the obtained cell suspension, fixed in methyl or ethyl alcohol, dried, stained according to Romanovsky-Giemsa. Lod microscope examines iOO lymphocytes, noting among them

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the number (%) of cells fixed on themselves three or more red blood cells (ROCK). If the content of lymphocytes in the patient's blood (%) and the total number of leukocytes are known, the absolute content of ROCK in 1 ml of blood is calculated.

The treatment of sheep erythrocytes in glutaraldehyde is carried out as follows: 1 ml of 5% suspension of carefully washed native sheep erythrocytes is combined with a 0.25% solution of glutaraldehyde in a 1: 1 ratio. The mixture was kept at 37.0 ° C for 15 minutes, after which the erythrocytes were washed twice from the aldehyde with a buffered isotonic solution of sodium chloride pH 7.2 and adjusted to the initial volume with the same solution (5% suspension). To sensitize red blood cells, a serotonin solution is prepared with a concentration of 0.1 mg / ml in a buffered saline solution pH 6.4. 1 ml of 5% suspension of glutarized erythrocytes is centrifuged, the supernatant is removed, and 1 ml of serotonin solution is added to the erythrocyte sediment. The mixture was shaken thoroughly and kept at room temperature for 25-30 minutes, after which the erythrocytes were washed twice by centrifugation at 1500 rpm for 10 minutes with isotonic sodium chloride solution and brought to 0.5% suspension.

The method is illustrated by the following examples.

Example 1. Patient GO., 33 years old, re-entered the clinic of nervous diseases with complaints of weakness in the legs, staggering when walking, dizziness, dysfunction of the pelvic organs. From the anamnesis it is known that he has been sick since the age of 18, the course of the disease is remitting, exacerbations were noted in 1976, 1979, 1982, 1986.

On examination, it was noted that there was no pathology on the part of the internal organs, a pulse of 68 beats. in 1 min, blood pressure 120/80 mm Hg

Neurologically: horizontal nystagmus in both directions, speech is somewhat dizarthric. Spastic tetra-paresis with pathological reflexes. Abdominal reflexes are not caused. Pronounced static and dynamic ataxi. There is no pathology on the survey craniogram. The fundus of the eye: decoloraci temporal halves of the optic nerve disks. Diagnosis: multiple sclerosis, cerebrospinal form.

The clinical diagnosis of multiple sclerosis is confirmed immunologically. The reserpine test positive number of serotonin rosettes (9%) practically did not increase (2%) after administration of reserpine. This suggests that the immune system did not give the usual, characteristic for healthy individuals, response to endogenous serotonin in the form of an increase in the number of immune outlets.

PRI mme R 2. Patient V., 27 years old, was admitted to the clinic for nervous diseases for the first time 7.V.37 complaining of reduced vision, weakness in the right leg, numbness of the fingers, a shaky gait. In Nv 1987, he was treated in the clinic for eye diseases with a diagnosis of retinal neuritis. After three weeks, my eyesight was partially restored, but weakness in the legs joined, more pronounced in the right, staggering when walking. On examination: pathology on the part of the internal organs was not revealed; pulse 72 beats. in 1 min, blood pressure 20/75 mm Hg Neurologically: the patient is euphoric, criticism is reduced. Gait atactic, strength in the legs is reduced - 3.5 points. Tendon jerks are quickened to the right higher than left. Pathological reflexes from two sides. Abdominal reflexes are not caused. Static and dynamic ataxi. There is no pathology on craniogram. The fundus of the eye: blanching of the temporal halves of the optic nerve discs The diagnosis was made: multiple sclerosis, cerebrospinal form, debut of the first attack. The clinical diagnosis is confirmed immunologically. The reserpine test is positive: the number of serotonin sockets (7%) did not increase after administration of reserpine (2%). Immunological diagnosis: multiple sclerosis.

Conducting a reserpine test in 1 patients with multiple sclerosis (cerebrospinal and stem-cerebellar forms) showed that, in all, administration of reserpine did not increase the number of immune to serotonin ROCK: before the administration is more harmful, 135 ± 13, after administration, 39.7 ± 13.7, 01

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The formulation of the reserpine test in patients with other types of lesions of the nervous system confirmed the specificity of the method.

ROCK before the introduction of reserpine 9.1 ± 0.9, after the introduction of 44.3 ± 2.7,, 01.

Healthy donors (20 people) responded to the administration of reserpine by a sharp increase in the number of serotonin ROCKs immune to blood,%: before the administration of 6.7 ± 0.3, after the administration of reserpine, 37.3 ± 2.1.

Thus, the diagnostic accuracy of the proposed method reached 100%, while the diagnostic accuracy of autoantibodies to brain tissue was 77-82%.

Claims (1)

Formula of the invention A method for diagnosing multiple sclerosis by taking blood and then immunoassaying it, characterized in that, in order to improve the diagnostic accuracy, reserpine is administered to the patient after the blood is taken, and after 30 minutes a second blood sample is taken, 0.1 are added to both samples. -0.2 ml of a 0.5% suspension of erythrocytes of a ram loaded with serotonin are incubated for 10-15 minutes at 37, C, the number of rose-forming lymphocytes is counted in a smear and in the absence of a significant increase in the index between both samples ruyut multiple sclerosis.