SU563964A1 - Method for evaluation of viral hepatitis seriousness - Google Patents

Description

one

The invention relates to medicae and can be used to estimate stress and predict severe forms of viral hepatitis.

A known method for the diagnosis of viral hepatitis, consisting in the final activity of the enzyme fructose-1 phosphagaldolase in serum of patients with iX

However, the known method does not provide a high level of analysis.

also the possibility of earlier diagnosing the disease and predicting its course,

In order to increase the speed of analysis, as well as the possibility of earlier diagnosing the disease and predicting its course according to the proposed method, blood serum is treated with shutein, then 250-500 J of substrate - glycyl-phenylalaniline-nitroanilide is added to the sample at room temperature flow min

The method for assessing the severity of viral x epatite is as follows.

The activity of the enzyme-catheps1psh with a CEM of the disintegrating l lOrOj5-Ly is determined. i) (: iv iffc; -pause the item iste nol, and then c-ucjijnTu ;. - gly1щ-fennlalanil r nitroishpdol, irifxy ban the solution with the subsequent definition of the D spectral C spectra) piniecKJ. Substrate taken in tsopichestepa 2 500 about 500 per sample. Incubation of tg; 1by peout at 37 ° C for 20 min.

Example. To 0.25 mp of serum in the experimental iipoGiipi: o add ml of rastran cine theine (to activate the enzyme) in adegatively 0.1 M 5.5 buffer (cysteine is dissolved at the rate of 17 mg in 1 ml of buffer) and maintain 5 min at room temperature. Diluted with 1-synthetic substrate - glyshch5L-11S1P1lalan1,; 1 -.- p nltroann.shd in the same buffer at the rate of 250 5 of the substrate in 0.75 ml of buffer. 0.75 ml of substrate solution in buffer is added to the test tube. Pre-dose volume up to 3 ml shstyk buffers. All the same is poured into the control tube, only clean is added instead of the substrate. The test tube is viewed against a control tube. On an SF-4A spectrophotometer with a BOJHIBI length of 110 MMK. Behind-

The onbiTss Jo., j and control tubes were placed in a t-thermostat at 37 ° C for 20 minutes (it was established experimentally that the greatest activity of the enzyme was manifested during this time). After incubation, samples are re-examined. From the second, larger vsppa, the first will be removed. The resulting axicstic value is converted to y- / mg using a calibration curve. The resulting value is cleared by 4 (in terms of 1 m serum). This is obtained if the p-nitroanipine if, which ottenil cathepsin C from 1 ml syvo-,

The number of surveyed

k) diseases of patients

18

Easy

27

Sre bottom t

20

T wish

Rota from substrate - glycyl-phenylalanyl-p-nitroanilio for 20 min of incubation. The calibration curve is constructed for p-nitroanipine. In healthy children, cathepsin C is not detected in the serum (is equal to O.) In viral hepatitis in children, especially when it is healthy, its activity is cathepsin G: in blood serum it reaches 12 / MP.

In tab. Figure 1 shows the results of an increase in cathepsin C activity in the acute period of the disease, depending on the severity of the process.

The multiplicity of increase in enzyme activity. ()

2.5 times

3.5 times

5.5 times

The use of the proposed method for determining the activity of cathepsin C in the serum of children with viral hematatitis simplifies the procedure by determining the enzyme activity in the serum; the possibility of an early assessment of the degree of severity of the pathological process in the liver and the timely appointment of appropriate therapy; as well as the ability to predict the course of the disease, which greatly helps in the treatment of viral hepatitis, especially severe forms. Formula of the invention A method for assessing the severity of viral hepatitis by determining the activity of serum fermetto in the blood serum spectrophotometrically, characterized in the specificity of the analysis, as well as the possibilities of earlier diagnosing the disease and predicting its course, the serum is treated with cyotine, then 2 50-500 is added to the sample. Substrate is glycyl-phenipalanil -p-iitroaipida, incubate the mixture at room temperature for 20-25 minutes. Sources of information taken into account in the examination: 1. J. Laboratory work, 1955, No. 6, 7, method of I. I. Tovarkitsky and E. N. Valuiskaya.