RU2101703C1 - Method for detecting the cases of inflammatory process activity in rheumatoid arthritis patients - Google Patents

Abstract

FIELD: medicine. SUBSTANCE: method involves carrying out blood serum deproteinization, obtaining proteinless sample and determining its optical density by using spectrophotometer. The sample is diluted with distilled water in 1: 9 proportion, solution optical density is determined at 254 nm wavelength. The value is used for making conclusions concerning medium mass molecules contents. The value being greater than 0.260 optical units, inflammatory process activity is considered to be the case. EFFECT: simplified method.

Description

 The invention relates to medicine and is intended to assess the activity of the inflammatory process in rheumatoid arthritis by means of a biochemical study of blood serum.

 Known methods for diagnosing the activity of the inflammatory process in rheumatoid arthritis by determining serumcoid [1] sialic acids [2] proteinograms [3] in the blood serum and others. These methods are laborious and difficult to implement.

 There is also a method for determining the presence of inflammatory-destructive process activity in rheumatoid arthritis by determining the level of connective tissue metabolites - glycosaminoglycans in blood serum [1]. The method includes serum proteolysis by papain, subsequent incubation, deproteinization, obtaining a protein-free sample by centrifugation, stepwise sedimentation, sample cleaning and determination of its optical density on a spectrophotometer at a wavelength of 523 nm. The optical density is used to calculate the quantitative content of glycosaminoglycans, and if it is greater than 0.78 g • 10 / l, the presence of an inflammatory-destructive process is diagnosed.

 The disadvantage of this method is its duration, multi-stage, the need for imported expensive reagents. The study takes 3 days.

 The objective of the invention is the acceleration and simplification of the method.

 The problem is solved in that in a method for determining the activity of the inflammatory process in rheumatoid arthritis, including deproteinization of blood serum, obtaining a protein-free sample and determining its optical density on a spectrophotometer, the sample is diluted with distilled water in a ratio of 1: 9, the optical density of the solution is determined at a wavelength of 254 nm, according to its value, the content of molecules of medium weight is judged and in the presence of the latter more than 0.260 optical units, the presence of inflammatory percent activity is diagnosed CCA.

 Determining the content of medium-weight molecules by diluting a protein-free sample with distilled water in a ratio of 1: 9 and determining the optical density of the resulting solution on a spectrophotometer at a wavelength of 254 nm allows you to speed up and simplify the method, as it eliminates such lengthy steps as proteolysis, incubation, and sample purification. The method for determining the presence of inflammatory activity by the content of medium-weight molecules takes 30 minutes.

 The method is as follows: produce a blood sampling, centrifuged it to obtain serum. To precipitate coarse proteins, deproteinization is carried out by adding 0.5 ml of a 10% trichloroacetic acid solution to 1 ml of blood serum and mix. To obtain a protein-free sample, centrifugation is carried out at 3000 rpm for 20 minutes. The obtained protein-free sample is diluted with distilled water in a ratio of 1: 9 and the optical density of the resulting solution is determined on a spectrophotometer at a wavelength of 254 nm. The optical density is used to judge the level of medium-weight molecules. The content of medium-mass molecules is expressed in units quantitatively equal to the optical density. In practically healthy people, it is 0.240 + 0.02 optical units. The content of molecules of average weight greater than 0.260 units of optical density suggests the presence of inflammatory activity in rheumatoid arthritis.

 Example 1. Patient F., 54 years old, was admitted with a diagnosis of rheumatoid arthritis, polyarthritis. The patient was taken blood from a vein in an amount of 4 ml, on an empty stomach. Blood was centrifuged, serum was obtained. To precipitate coarse proteins, 0.5 ml of trichloroacetic acid was added to 1 ml of blood serum, shaken, and centrifuged at 3000 rpm for 20 min in a clinical laboratory centrifuge (OPn-3). We obtained a protein-free sample and diluted it in a ratio of 1: 9, for which 4.5 ml of distilled water was added to 0.5 ml of the supernatant. The optical density of the samples was determined on a spectrophotometer (SF-46) at a wavelength of 254 nm. Its value and, accordingly, the level of molecules of average mass amounted to 0.375 optical units, which exceeded 0.260 optical units and indicated the presence of inflammatory activity. Clinical and laboratory examination using laborious generally accepted biochemical research methods also revealed the presence of inflammatory activity. Final diagnosis: rheumatoid arthritis, polyarthritis, stage III, functional insufficiency I-II, degree of activity III.

 The patient was shown surgical treatment for synovcapsulectomy. Such operations are preferably done during the period of remission or with a minimal degree of activity of the inflammatory process in order to avoid postoperative complications. Therefore, the patient underwent conservative treatment with non-steroidal anti-inflammatory drugs, intra-articular introduction of the Kenalog, and physiofunctional treatment aimed at reducing the activity of the inflammatory process. After the course of therapy, the presence of activity of the inflammatory process by the proposed method was re-conducted. The level of medium-weight molecules decreased to 0.284 optical units, i.e. the activity of the inflammatory process was reduced to a minimum. Surgery was performed. The postoperative period was uneventful, the patient was discharged home in satisfactory condition for outpatient treatment.

Example 2. Patient T., 14 years old, was admitted with a diagnosis of rheumatoid arthritis. The patient had a determination of the level of medium-weight molecules in blood serum by the proposed method. It amounted to 0.242 optical units, which indicated the absence of inflammatory activity. A generally accepted clinical, laboratory, including biochemical, examination (determination of sialic acids, seromucoid, proteinograms, etc.) was also carried out, in which the presence of inflammatory activity was also not revealed. The final diagnosis was made: rheumatoid arthritis, oligoarthritis, seronegative, stage I, functional failure I, degree of activity 0-1
The patient was shown synovcapsulectomy of the left knee joint, which was performed without a preliminary course of conservative drug and physiological treatment. The postoperative period was uneventful, the patient was discharged for outpatient treatment at the place of residence.

 Using the proposed method, 68 patients with reliable rheumatoid arthritis were examined at the Republican Arthrological Center at SarNIIITO. The method allows to determine the presence of activity of the inflammatory process in both the initial and far advanced stages of the disease.

 The proposed method can significantly simplify the assessment of the activity of the inflammatory process in rheumatoid arthritis, due to a significant reduction in study time, and dispense with expensive imported reagents.

Sources of information
1. Lempert M.D. Biochemical research methods. Kishenev, 1968

 Laboratory research methods in the clinic: Handbook / Ed. Menshikova V.V. M. Medicine, 1987, 368 pp.

 Sigidin I.A. Guseva N.G. Ivanova M.M. Diffuse connective tissue diseases. A guide for doctors. M. Medicine, 1994.554 s.

 Karyakina E.V. Zhadenov I.I. Glycosaminoglycans of blood serum and synovial fluid as an indicator of the inflammatory and destructive process in rheumatoid arthritis. Saratov, 1986 p. 4-6 (prototype).

Claims (1)

 A method for determining the presence of inflammatory activity in rheumatoid arthritis, including deproteinization of blood serum, obtaining a protein-free sample and determining its optical density on a spectrophotometer, characterized in that the protein-free sample is diluted with distilled water in a ratio of 1 to 9, the optical density of the solution is determined at a wavelength of 254 nm, by its value, the content of medium-weight molecules is judged and, with a value of the latter greater than 0.260 optical units, the presence of inflammatory activity is diagnosed processes.