RU2706733C1 - Diagnostic method of molecular phenotypes of osteoarthritis - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to medicine, particularly to laboratory diagnostics, rheumatology, orthopedics and traumatology. A diagnostic technique for molecular phenotypes of osteoarthritis involves plasma analysis of venous blood by enzyme immunoassay to concentration of a factor induced by oxidative stress 1, and interleukin 1, wherein if the concentration of only the factor induced by oxidative stress 1 is increased, an oxidative phenotype of osteoarthritis is diagnosed, if the concentration of only interleukin 1 increases, the inflammatory osteoarthritis phenotype is diagnosed, with simultaneous increase or normal indices of the oxidative stress inducing factor 1 and interleukin 1, a mixed inflammatory-oxidative phenotype of osteoarthritis is diagnosed.

EFFECT: invention can be used for diagnosing molecular phenotypes of osteoarthritis.

1 cl, 2 ex

Description

The invention relates to medicine, in particular to laboratory diagnostics, rheumatology, orthopedics and traumatology, in particular to a quantitative study of the concentration of biological molecules in venous blood and the assessment of their ratios.

A known method for the diagnosis of osteoarthritis and other diseases of connective tissue, including determining by Western blotting levels of fragments of the cleavage of alpha-1 protein and collagen IX chain in blood and synovial fluid. The result is the concentration, the magnitude of which is judged on the presence of connective tissue disease, in particular osteoarthritis (application No. 2009136441).

However, the known method is expensive, has low specificity for the diagnosis of osteoarthritis, as it is used, in general, for connective tissue diseases, the sector of which is wide, not intended for the diagnosis of phenotypes of osteoarthritis.

A known method for the differential diagnosis of osteoarthritis and rheumatoid arthritis, comprising measuring the concentration of the peptide of the C1 chain of protein 2 of the intermediate layer of cartilage. The result is the concentration of the peptide of the C1 chain of protein 2 of the intermediate layer of cartilage, the high level of which judges the presence of osteoarthritis (patent No. 2603484).

However, the known method has low specificity for the diagnosis of osteoarthritis, as it evaluates only cartilage changes, does not take into account bone remodeling, is not intended for the diagnosis of phenotypes of osteoarthritis.

As a prototype, a method for the diagnosis of arthritis and osteoarthritis based on the determination of at least three diagnostic agents in saliva selected from the group consisting of gamma-interferon induced by interferon protein-10, interleukin-2 and total protein of saliva was selected. When determining elevated concentrations of these molecules, osteoarthritis is diagnosed. (patent No. 2603484).

However, the known method is associated with the determination of biological molecules in saliva, the composition of which is significantly different from the composition of the blood, does not take into account the ratio of molecular concentrations, is not intended for the diagnosis of phenotypes of osteoarthritis.

The objective of the present invention is a laboratory study of concentrations in venous blood by enzyme-linked immunosorbent assay of factor induced by oxidative stress 1 and interleukin 1 to determine the phenotype of soteoarthritis.

The presented problem is solved in that in a method for diagnosing phenotypes of osteoarthritis, including a laboratory study of concentrations in venous blood by enzyme-linked immunosorbent assay of factor induced by oxidative stress 1 and interleukin 1, characterized in that if the concentration of only the factor induced by oxidative stress 1 is diagnosed, the oxidative phenotype of osteoarthritis is diagnosed, with an increase in the concentration of only interleukin 1, the inflammatory phenotype of osteoarthritis is diagnosed, while enii normal parameters or factor induced oxidative stress 1 of interleukin 1 and diagnose mixed-oxidative inflammatory osteoarthritis phenotype.

It is advisable to determine the increase in the concentration of the factor induced by oxidative stress 1 and interleukin 1 to use the reference interval of normal values determined by the manufacturer of the reagents for enzyme immunoassay, consider the concentration increase above the 99th percentile of the refrain interval.

The method is as follows.

Venous blood is drawn into a test tube, which is then placed in a centrifuge and centrifuged for 5 minutes at a speed of 1000 rpm. The resulting blood plasma is taken in any known manner and placed in an Eppendorf type storage tube. Thus obtained venous blood plasma is studied using enzyme-linked immunosorbent assay in any known manner. To determine the concentrations of the factor induced by oxidative stress 1 and interleukin 1, ready-made commercial reagent kits are used. The analysis is carried out in accordance with the manufacturer's instructions.

When determining the concentration of the factor induced by oxidative stress 1 above the 99th percentile of the reference interval and the normal concentration of interleukin 1, the oxidative phenotype of osteoarthritis is determined.

When determining the concentration of interleukin 1 above the 99th percentile of the reference interval and the normal concentration of the factor induced by oxidative stress 1 and interleukin 1, the inflammatory phenotype of osteoarthritis is determined.

When determining the concentrations of the factor induced by oxidative stress 1 and interleukin 1 within or above the 99th percentile of the reference interval, a mixed phenotype of osteoarthritis is determined.

Clinical example of the method.

Example 1

Patient A., 56 years old, with complaints of pain in the mechanical rhythm of the right knee joint.

Clinical and radiological examination made it possible to establish a diagnosis of osteoarthritis of the right knee joint. To select the patient’s treatment tactics, it was decided to determine the molecular phenotype of the disease. For this, blood was drawn from the cubital vein using a vacuum system into a test tube. After centrifugation and obtaining blood plasma, an enzyme-linked immunosorbent assay was performed using commercial kits to determine the factor induced by oxidative stress 1 and. The results are obtained: the concentration of the factor induced by oxidative stress 1-4.16 ng / ml (normal: 0.56-1.85 ng / ml), interleukin 1-1.02 pg / ml (normal: 0.00-5.00 pg / ml). Given the increased concentration of the factor induced by oxidative stress 1 and the normal concentration of interleukin 1, the oxidative phenotype of osteoarthritis was determined. The patient is prescribed appropriate pathogenetic therapy.

Example 2

Patient B., 64 years old, with complaints of pain of a mechanical rhythm in the left knee joint.

Clinical and radiological examination made it possible to establish a diagnosis of osteoarthritis of the left knee joint. To select treatment tactics, the patient decided to determine the molecular phenotype of the disease. For this, blood was drawn from the cubital vein using a vacuum system into a test tube. After centrifugation and obtaining blood plasma, an enzyme-linked immunosorbent assay was performed using commercial kits to determine the factor induced by oxidative stress 1 and. The results were obtained: the concentration of the factor induced by oxidative stress 1 - 0.98 ng / ml (normal: 0.56-1.85 ng / ml), interleukin 1 - 10.69 pg / ml (normal: 0.00-5.00 pg / ml). Given the increased concentration of interleukin 1 and the normal concentration of the factor induced by oxidative stress, the inflammatory phenotype of osteoarthritis was determined. The patient is prescribed appropriate pathogenetic therapy.

Information sources:

1. RF patent RU 2603484 C2, 2016 (prototype).

2. RF application RU 2009136441 A, 2011.

Claims (1)

A method for the diagnosis of molecular phenotypes of osteoarthritis, including the determination in plasma of venous blood by enzyme immunoassay of the concentration of the factor induced by oxidative stress 1 and interleukin 1, characterized in that in the case of an increase in the concentration of only the factor induced by oxidative stress 1, the oxidative phenotype of osteoarthritis is diagnosed with an increase concentrations of only interleukin 1 diagnose the inflammatory phenotype of osteoarthritis, with a simultaneous increase or normal levels of Ktorov induced oxidative stress 1 of interleukin 1 and mixed diagnose inflammatory oxidative osteoarthritis phenotype.