SU1323083A1 - Method of diagnosis of hereditary spherocytosis - Google Patents

Abstract

This invention relates to hematology. The purpose of the invention is to improve the accuracy of the method. Samples are examined for the presence of hereditary spherocytosis and then (Na + K) ATP-ase activity and lipid composition of erythrocyte membranes are determined. With an increase in (Na + K) ATP-ase activity of erythrocyte membranes and an increase in the lipid fraction in the free fatty acid zone in combination with other negative tests for hereditary spherocytosis, the latent form of this disease is diagnosed. The method provides medical and genetic consultations.

Description

This invention relates to medicine and relates to a method for diagnosing hereditary spherocytosis.

The aim of the invention is to improve the accuracy of the method by detecting the latent form of the disease.

The method is carried out as follows.

In individuals with suspected hereditary spherocytosis, blood is taken from a finger and checked for microspherocytosis, a reduced diameter and a reduced osmotic resistance of erythrocytes and an increased number of reticulocytes. In parallel, we take blood in a volume of 15 ml, erythrocyte membranes are isolated by the Brinkman method, ((-K) ATP-ase activity, protein by Loury, phosphorus by Fiske, fractional composition of lipids by Stahl is determined.

In the presence of microspherocytosis, reduced diameter and reduced osmotic resistance of erythrocytes and an increased number of reticulocytes with a simultaneous increase () in ATP-ase activity of erythrocyte membranes and a visually detectable increase in the lipid fraction in the free fatty acid zone confirm hereditary spherocytosis. If the decrease in (Na + K) ATP-ase activity of erythrocyte membranes and an increase in the lipid fraction in the free fatty acid zone is combined with the absence of microspherocytosis, a decrease in diameter and reduced osmotic resistance of erythrocytes and an increased number of reticulocytes, a latent form of inheritance is diagnosed spherocytosis.

Example 1. A patient of 30 years old was admitted to the hospital with a diagnosis of hereditary spherocytosis. Complaints of general weakness, feeling of body in the left hypochondrium, subfebrile temperature. Sick since childhood.

Objectively: the skin and the visible mucous jaundices, the spleen protrudes 7 cm from the hypochondrium, dense consistency, the projection of the gallbladder is painful.

Blood test НЬ 94 g / l; er 2.8 "10 / L; color indicator 1; ESR 4 mm; lake 5.0-10 / l; thrombus. 47%; retic. 78%; basophilic reticulum c. a thick drop +++; the average diameter of erythrocytes is 7.0 microns; osmotic pe5

five

50

five

0

five

red blood cell resistance O, 56-0.32% NaCl. A moderate erythrocyte anisocytosis due to saturated micro and not a large number of macrocytes is noted, small polychromasis, ovalocytes 5–8 are found in the visual field, as well as single tears. Leukocyte formula,%: fell. 6.0; segm 50.0; my. 5.0; zoz 1.0; limf 38.0.

Functional liver tests: total bilirubin 46.5 µmol / l, direct 7.2 µm mol / l, free 39.3 µm mol / l, thymol sample 9.0, total protein 7.2 g / l.

A special study of erythrocyte membranes revealed that (Na-t-K) ATP-asta activity was 2.21 mM / mg ih, the lipid fraction in the free fatty acid zone was visually increased.

The patient produced splenectomy and cholecystectomy.

Blood test after surgery: HB 120.0 g / l; er 4.2 10 / l; color index 0.91; erythrocyte morphology did not change, but erythrocytes with the Jolly body appeared.

Based on these studies, erythrocyte membranes confirmed the diagnosis of hereditary spherocytosis.

Example 2: Patient B. spent 6 years at the clinic with a diagnosis of congenital hemolytic anemia (microspherocytosis is suggested). Complaints of general weakness, high temperature, drowsiness and restlessness. The child has been sick since the age of one.

Objectively: pronounced jaundice of skin prokrov and visible mucous membranes, herpes on the lips, spleen protrudes 7 cm from the hypochondrium, 6 cm from the liver.

Blood test: Hb 90.0 g / l; er 2.8-10 / l, color indicator 0.93; ESR 22 mm; lake 6,810 / l; thrombus. 230.4 U / l; retic. 110%; Bacophilus netting in a thick drop ++++; the average diameter of erythrocytes is 7.0 µm, the osmotic resistance is 0.56– 0.32 NaCl. There is marked anisocytosis due to micro- and macrocytes, small poikilocytosis, part of erythrocytes hypochromic, part polychromatophilic, ovalocytes are found (20-25 in the visual field, including narrow ellipsoid, single tear-shaped, Lecoicocyte form, I-form, Iryomas, and narrow ellipsoid, single tear-shaped, Lecoicocyte form, I-form, Iryomas, and narrow ellipsoid, single tear-like, Lecoicocyte form, I-form, and I-form, Leococytoma form, I-20, 25). 11.0;

segm 38.0; my. 3.0; eos. 6.0; limf 42.0.

In cytological examination of a peripheral blood smear, red blood cells containing Hb 10%. When blood was incubated with cresyl brilliant blue dye, intraerythrocyte inclusions of unstable hemoglobin were not detected.

Functional treatment of cure: total bilirubin due to an indirect reaction of 25.2 µmol / L; timolovy test 6; mercury sublimate reaction 92%.

Urine and feces without change.

In terms of microcytosis, small diameter and low resistance of erythrocytes, hereditary spherocytosis is suspected. As a result of special studies of erythrocyte membranes, it turned out that () ATP-asta activity was reduced (0.39 mM / mg / hr), the lipid fraction in the free fatty acid zone was not visually increased, and therefore the diagnosis of inherited spherocytosis was not increased. excluded.

Treatment with blood transfusions, vitamins B ,, Bg, folic acid.

The patient was transferred to the surgical department, where splenectomy was performed. The patient was discharged with a diagnosis of nonsferocytic hemolytic anemia.

PRI me R 3. Patient V. 60 years.

Subjectively: no complaints.

Objectively: the patient is of a proper physique, the subcutaneous fat is poorly developed, the liver and spleen are not enlarged.

Blood test: HB 155 g / l; er

ten

4,910 / l, color indicator 0,95 ESR 5 mm; lake 5.2-10 / l; thrombus. 54%; retic. 5%, basophilic reticulation in a thick drop +. Small anisocytosis due to single micro and macrocytes is noted. Leukocyte formula,%: fell. 2.5; segm 59.5; my. 3.5; eos. 2.0; bases. 1.0; limf 31.5.

By all measures, the patient was found to be practically healthy. However, due to the fact that his children have a well-established hereditary Spherocytosis, a special study of the erythrocyte membranes has been carried out. It turned out that () ATP-asta membrane activity was increased and was 2.2 mM / mgh. Lipid fractions in the free fatty acid zone are visually increased.

As a result of research, the patient was diagnosed with a latent form of hereditary spherocytosis.

Example 4. Patient G., aged 8, entered the children's ward during a crisis with complaints of general weakness and yellowness.

Objectively: the yellowness of the skin and visible mucous membranes is noteworthy.

Blood test: HB 93.0 g / l; er 2.7 1 O / l; color indicator 1, 0, retic. 270%. The erythrocyte tendency to marocytosis is noted, die} P1a polychromasy.

Presumptive diagnosis: hereditary spherocytosis. The shape, diameter, and resistance of erythrocytes were determined. It turned out that microspherocytosis of erythrocytes is absent, diameter 7.5 μm, resistance 0.48 0.28. When conducting an additional special study of erythrocyte membranes, it turned out that (Na + K) ATP-asla activity is within the normal range, the lipid fractions in the free fatty acid zone are not visually increased. The diagnosis of hereditary spherocytosis is not confirmed by the result-1i and studies.

The patient studied the activity of the enzyme glucose-6-phosphate dehydrogenase (G-6-PD), it turned out that the activity of the enzyme G-6-PD was O me / g Pv.

On the basis of these data, a diagnosis was made: hereditary hemolytic anemia caused by a deficiency of the enzyme G-6-PD.

Blood test: Hb 103 g / l; er 4.1 U / l.

The patient was discharged in a satisfactory condition.

The method has been tested in laboratories of biochemistry, as well as hereditary and acquired anemias in clinical and outpatient patients of the Scientific and Research Institute of Hematology and Blood Transfusion.

The method provides 100% accuracy of diagnosis of the disease, including latent forms, which allows timely targeted treatment, and also makes it possible to correctly predict the disease.

513230836

in the progeny and to conduct medical genes of the disease, in addition olredetic counseling. lll (Na. + K) apt-az activity

and lipid composition of membranes eritrotsiFormula invention comrade and in the absence of reducing dia5metra and reduce osmotic rezisSposob diagnostic nasledstvennogotentnosti erythrocytes amid uvelisferotsitoza by determining the shape, Cheney reticulocyte count in sodiametra, osmotic resistance, coupled with an increase () AFTti erythrocytes and the number of the reticulo-ase activity and increase lipid cells, characterized by the Ono fraction in the free fatty zone

that, in order to improve the accuracy of the acidity relative to the diagnostic norm by detecting a latent form, the latent form of the disease is formed.

Claims (1)

Claim A method for the diagnosis of hereditary spherocytosis by determining the shape, diameter, osmotic resistance of red blood cells and the number of reticulocytes, characterized in that, in order to improve the accuracy of the method by detecting latent form of the disease, they additionally determine (Na + K) ATPase activity and lipid composition erythrocyte membranes and in the absence of a decrease in diameter and a decrease in the osmotic resistance of red blood cells against the background of an increase in the number of reticulocytes in combination with an increase in (Na ⁺ + K *) Α Τ phase activity and An increase in the lipid fraction in the free fatty acid zone relative to the norm diagnoses the latent form of the disease.