RU2312360C1 - Method for screening-diagnosing liver fibrosis cases using salivary biochemical tests - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method involves determining iron concentration in mcm/l, gamma-glutamyltranspeptitase in mccat/l and ornitindecarboxylase in ncat/ml. Combined iron concentration increase higher than 1.0 mcm/l, gamma-glutamyltranspeptitase higher than 0.2 mccat/l and ornitindecarboxylase higher than 0.05 ncat/ml in given patient, indicates liver fibrosis presence.

EFFECT: high accuracy in diagnosing liver fibrosis in early stage.

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Description

The invention relates to medicine and can be used in gastroenterology for the early diagnosis of fibrotic processes developing against the background of chronic liver diseases (CKD) of various etiologies.

The actual problem of gastroenterology is to increase the efficiency of early diagnosis of fibrotic changes in the liver with its chronic pathology, which is due to the high prevalence, increase in morbidity, mortality and increased economic costs for the treatment of liver cirrhosis and its complications, as well as for liver transplantation worldwide. The main forms of CKD are alcoholic liver disease, non-alcoholic steatohepatitis and chronic hepatitis C. As you know, the treatment of liver cirrhosis (CP), which develops in the final stage of these diseases, is often ineffective. However, at the initial stages (fibrosis), this process can still be influenced, suspended. This implies the importance of early detection, dynamics assessment and (if possible) elimination of the triggering factor of fibrogenesis.

Until recently, a liver biopsy was universally considered the gold standard in assessing fibrosis. However, this method, due to the so-called "hit errors", can give a false idea about the nature of changes in the liver. In addition, there is always a risk of developing severe complications of this invasive diagnostic procedure. (Massimo Pinzani (Florence, Italy). The evolution of liver fibrosis: from hepatitis to cirrhosis. // Russian Journal of Gastroenterology, Hepatology and Coloproctology. - 2002 - No. 5.)

A new method for assessing fibrosis is the measurement in the systemic circulation of its serum markers, in particular the components of the intracellular matrix (VKM) (Galimova S.F., Nadinskaya M.Yu., Mayevskaya M.V., Ivashkin V.T. New data on the diagnosis and course of liver fibrosis. // Russian Journal of Gastroenterology, Hepatology and Coloproctology. - 2001 - No. 4. - pp. 22-28) - also has limited use, which is associated with the rather low specificity of most indicators for liver tissue and their high cost definitions. This method was taken by us as a prototype. In addition, there is evidence of the use of routine biochemical blood parameters as serum fibrosis markers, which indirectly indicate changes in various liver functions, in particular, protein synthesis. One of them is whey. (Bychkova V.I., Smirnov B.M., Lesnichuk L.V. Biochemical parameters of connective tissue in the diagnosis of the initial stage of liver fibrosis. // Clinical laboratory diagnostics. - 2003 - No. 1. - pp. 10-14. )

The study of saliva has several advantages compared with routine laboratory diagnostic methods using blood taken from a vein or from a finger. First of all, this is the simplicity and convenience of collecting material, the non-invasiveness and painlessness of this procedure, the absence of the risk of infection, and the possibility of multiple sampling. (Denisov A.B. Salivary glands. Saliva. M., 2003)

The aim of the invention is to develop an economical and non-invasive method for the early diagnosis of liver fibrosis, suitable for conducting extensive screening studies.

The goal is achieved by determining the content of iron, gamaglutamyltranspeptidase (GGTP) and ornithine decarboxylase (ODK) in the saliva of patients with CKD with confirmed liver biopsy fibrosis and conditionally healthy individuals. Iron was chosen by us as a likely indicator of changes in the protein synthesizing function of the liver in the presence of a fibrotic process. In the liver, transferrin is synthesized - a protein involved in the transport of iron. GGTP is a marker of cholestatic and parenchymal liver lesions. An increase in its level in the body’s media can be partly due to the induction of microsomal enzyme activity. (S. Sherlock, J. Dooley. Diseases of the liver and biliary tract. // M .: "GEOTAR-MED", 2002) UEC - an enzyme involved in the formation of biogenic polyamines, an increase in its activity is one of the earliest events, occurring during the transition of cells from dormancy to active proliferation. (Kolpashchikov I.E., Shamin G.A., Lak G.A., Beilin S.B. Ornithine decarboxylase in the maturation, regeneration and adaptation of the intestine. // Gorky, 1987)

A method for diagnosing fibrosis in CKD is that they use complex quantitative laboratory criteria for the formation of fibrosis (iron content, GGTP and ODK in saliva) in conditionally healthy individuals in screening, which differs significantly from determining blood fibrosis.

The proposed method is as follows. In a patient with CKD or a conditionally healthy person, in the morning on an empty stomach and before brushing your teeth, after preliminary rinsing of the oral cavity with water, saliva is collected in a conical tube. In saliva, the concentration of iron, GGTP and UEC is determined and compared with indicators calculated by comparing two samples (patients with CKD with fibrosis and conditionally healthy) using the Bayes formula and Wald's sequential analysis. (E.V. Gubler, 1978; V.I.Broitman, 1991)

With a combined increase in the salivary iron level of more than 1.0 μm / l, GGTP of more than 0.2 μkat / l, ODC of more than 0.05 nkat / ml, the examined person has a 95% probability that he has liver fibrosis.

The diagnostic effectiveness of the proposed method was studied in 20 patients with chronic liver disease (CKD) (chronic hepatitis C (CHC)). These patients made up group F - patients with fibrotic changes in the liver. The diagnosis of liver fibrosis in all cases was confirmed by puncture biopsy of the liver (PBP). The control group (K) consisted of 20 conditionally healthy individuals.

The iron content was determined using spectrophotometry (atomic sorption method), to determine the GGTP level, a unified method was used using the Vital-Diagnostics SPb reagent kit, the concentration of UEC was determined on a photocolorimeter by an enzymatic method based on the loss of substrate (ornithine) according to the method of V. Khramov . (V.A. Khramov. A simple method for determining the activity of ornithine decarboxylase in mixed saliva of a person. // Clinical laboratory diagnostics. - 1997 - No. 4. - P.14-15).

The results of the study of iron, GGTP and ODK in the saliva of patients with CKD and conditionally healthy individuals are presented in table. The average salivary iron level in group F was 1.67 ± 0.34 μm / L, and in the control group it was 0.91 ± 0.27 μm / L. The content of GGTP in the saliva of patients with liver fibrosis was 0.3 ± 0.04 μkat / L, of conditionally healthy - 0.19 ± 0.03 μkat / L. The concentration of UEC, according to the results of the study: in the saliva of the group F - 0.12 ± 0.03 ncat / ml, in the saliva of the group K - 0.04 ± 0.01 ncat / ml.

Thus, the combined increase of three indicators such as salivary iron (more than 1.0 μm / l), GGTP in saliva (more than 0.2 μkat / l), UEC in saliva (more than 0.05 nk / ml) in the subject screening, with a probability of 95% will indicate the presence of liver fibrosis.

Table Method for screening diagnosis of liver fibrosis according to salivary biochemical criteria Indicator F (n = 20) K (n = 20)

Iron, μm / L 1.67 ± 0.34 0.91 ± 0.27 * GGTP, mkkat / l 0.3 ± 0.04 0.19 ± 0.03 ** UEC, ncat / ml 0.12 ± 0.03 0.04 ± 0.01 * Note: * - significance of differences with the control group (according to the Wilcoxon-Mann-Whitney criterion) p <0.01, ** - significance of differences with the control group (according to the Wilcoxon-Mann-Whitney criterion) 0.01 <p <0.05 .

Claims (1)

A method for screening diagnosis of liver fibrosis according to salivary biochemical criteria, including the quantification of saliva, gamma glutamyl transpeptidase and ornithine decarboxylase in saliva, characterized in that when the combined increase in iron is more than 1.0 μm / l, gamma glutamyl transpeptidase more than 0.2 μkat / l and ornithine carboxy 0.05 nkat / ml in the subject diagnosed with liver fibrosis.