RU2475754C2 - Method of predicting gastroduodenal pathology course in children - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: before treatment venous blood is sampled, centrifuged with obtaining plasma in which levels of IgM to endotoxin and S aureus are determined by means of standard enzyme immunoassay. Number of performed H.pylori eradications, patient's sex are determined and prognostic probability is calculated by formula: P=1/(1+exp(-(X1*0.05-X2*0.08+X3*2.31+X4*1.06-2.99))), where P is prognostic level of probability; X1 is level of IgM to S.aureus; X2 is level of IgM to endotoxin; X3 is sex (f - 0, M - 1); X4 is number of performed H.pilori eradications. If P≥0.36, outcome is predicted, if P <0.36 favourable outcome is predicted.

EFFECT: method makes it possible to predict severity of gastrointestinal pathology in children in due time and in low-invasive way.

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Description

The invention relates to medicine, namely to children's gastroenterology, and can be used to predict the course of gastroduodenal pathology in children.

In the structure of gastroenterological pathology in children, diseases of the upper digestive tract prevail, including chronic gastroduodenitis and peptic ulcer of the duodenum. In recent years, an important role in the development and recurrence of erosive and ulcerative lesions of the gastric mucosa and duodenum is assigned to changes in the microecology of the gastrointestinal tract, which are characterized by excessive growth of mucous microflora and an increase in its aggressive properties [see Peptic ulcer, chronic gastritis and esophagitis in the aspect of dysbiosis of the esophagogastroduodenal zone / V.V. Chernin, V. M. Chervinets, V. M. Bondarenko, S. N. Bazlov. - Tver: LLC Triada, 2004. - p.96-108]. An accurate prognosis of an unfavorable recurrent course of chronic gastroduodenitis and duodenal ulcer based on the study of specific immunity to individual antigens of opportunistic microorganisms seems to be very relevant, because will allow you to determine the individual characteristics of the tactics of therapeutic measures.

There are various methods for predicting the course of gastroduodenal pathology in children. Most of them are invasive and relate to the prognosis of duodenal ulcer.

A known method for predicting the course of peptic ulcer of the duodenum in children by examining the mucous membrane of the duodenum with inflammation [see Zaprudnov A.M. Handbook of Pediatric Gastroenterology / A.M. Zaprudnov, A.I. Volkov, K.I. Grigoriev. - M .: Medicine, 1995. - p.237-238], including the study of goblet cells in the villi and enterocytes of crypts in a state of mitosis.

A known method for predicting the course of peptic ulcer by determining the number of Paneth cells per crypt of the duodenum, and with a value of 1.6 cells or less, relapse is predicted within the next 6 months [see Pat. No. 2162600, Russian Federation, IPC G01N 33/48. A method for predicting the course of peptic ulcer in children / Potekhin P.P., Pereslegina I.A., Shirokova N.Yu., Sokolova I.L., Saralov S.N., Zhukova E.A., Artemova A.V. .; applicant and patent holder Nizhny Novgorod Research Institute of Pediatric Gastroenterology. - No. 99112392/14; declared 06/07/99; publ. 01/27/2001].

However, the above methods require morphological studies of the mucosa of the gastroduodenal region, are invasive, which limits their use in pediatric practice, especially in an outpatient setting.

A known method for predicting the risk of development of destructive changes in the mucous membrane of the gastroduodenal zone, in which the absolute number of peripheral blood neutrophilic granulocytes positive for SB80 receptors is determined, and provided this value is above 999 cells / ml, there is a risk of developing destructive changes in the mucous membrane of the gastroduodenal zone [see . Pat. No. 2391915, Russian Federation, IPC АВВ 10/00, G01N 33/53. A method for predicting the risk of developing destructive changes in the mucous membrane of the gastroduodenal zone / Nesterova IV, Shvydchenko IN, Kapranova NP, Bykovskaya E.Yu., Nikolaev VV; applicant and patent holder State Healthcare Institution "Center for Functional Surgical Gastroenterology" of the Department of Health of the Krasnodar Territory. - No. 2008148414/14; declared 12/08/2008; publ. 06/20/2010]. However, this method, determining the presence of destructive changes in the mucous membrane of the stomach and duodenum at the time of examination, does not allow to predict the further course of the disease.

The closest in technical essence to the claimed method is a method for assessing the risk of developing peptic ulcers, which includes the steps of quantifying the concentration of pepsinogen I (PGI) and gastrin-17 in a serum sample, in which by quantifying the concentration of pepsinogen I and gastrin-17 in a sample of the serum of the indicated individual, the choice of a method-specific reference value and cut-off value for the corresponding analytes, comparing the concentrations thus determined epsinogen I and gastrin-17 with a method-specific reference value and cut-off value, as a result of which the serum concentration of pepsinogen I and gastrin-17 is higher than the upper limit of the corresponding reference value or the serum concentration of pepsinogen I is higher than the upper limit of its reference value in combination with the concentration of gastrin 17 within the reference range or below its cut-off value, indicates an increased risk of peptic ulcer in the indicated individual [see Application for the invention of the Russian Federation No. 2001132335/14, IPC G01N 33/573. A method for assessing the risk of developing peptic ulcers, which includes the steps of quantitatively determining the concentration of pepsinogen I (PGI) and gastrin-17 in a serum sample / Sipponen P., Hjarkenen M., Suovaniemi O., Forsblom E.: stated. 04/28/2000; publ. 08/27/2003].

The disadvantage of this method is the need for repeated sampling of blood serum from an individual to determine the level of postprandial gastrin-17, which is not always technically feasible in children.

The aim of the present invention is the ability to predict the course of gastroduodenal diseases in children without morphological studies to determine individual tactics of patient management.

This goal is achieved by a method for predicting the course of gastroduodenal pathology by collecting venous blood in children with chronic diseases of the stomach and duodenum, in which the plasma obtained by centrifugation determines the level of Ig M to endotoxin and S. aureus using standard enzyme immunoassay, moreover, as an antigen endotoxin use Salmonella minnesota RE 595 glycolipid, S.aureus - ultrasonic disintegrate of these bacteria, determine the amount of H. pylori eradication and patient gender , subsequently, the prognostic probability is calculated by the formula: P = 1 / (1 + exp (- (X1 * 0.05-X2 * 0.08 + X3 * 2.31 + X4 * 1.06-2.99))) where P is the prognostic level of probability; predictors: XI - level of Ig M to S. aureus (μg / ml); X2 - level of Ig M to glycolipid (μg / ml); X3 - floor (w-0, m-1); X4 is the number of Helicobacter pylori eradications performed, with a probability level of P≥0.36, an unfavorable outcome is predicted (high risk of occurrence and relapse of erosion and ulcers in the gastric mucosa and duodenum), and with P <0.36 - favorable.

The method is as follows.

In fasting children, 2 ml of blood is taken from the median ulnar vein into a sterile pyrogen-free container, centrifuged and plasma is obtained. The sensitization of the plates with endotoxin and S.aureus antigens is carried out in phosphate-buffered saline with a pH of 7.2-7.4. Free areas are blocked with 0.1% bovine albumin.

Plasma samples are pre-diluted 300 times with a solution of phosphate-saline buffer with tween (FSB-T). Then 0.2 ml of each plasma sample is added to the plate. The control plasma with a known antibody content is pre-diluted 100 times and sequential double dilutions are made vertically on the control strip. After filling the plate with plasma samples, it is incubated at 37 ° C for 45 minutes. At the end of the incubation, the plate is shaken off and washed three times with a solution of FSB-T. 0.11 ml of IgM conjugate are added to each well of the plate. Incubated for an hour under the same conditions as the first stage. At the end of the incubation, the plates are washed three times with FSB-T solution and distilled water, after which 100 μl of a chromogen solution is added to each well. After a 20-minute incubation at 37 ° C, 50 μl of a 50% sulfuric acid solution are added to all wells of the plate to stop the reaction. The optical density is measured at a wavelength of 495 nm on a spectrophotometer.

The amount of IgM is determined from plasma titration curves with a known specific IgM content. Antibody concentration is expressed in μg / ml.

The analysis is carried out on standard equipment and standard reagents for enzyme immunoassay.

When questioning the patient, the number of previous H. pylori eradications is determined. The results of the determination of IgM for endotoxin and S.aureus, the number of H. pylori eradications performed and the gender of the patient are supplied in the formula:

P = 1 / (1 + exp (- (X1 * 0.05-X2 * 0.08 + X3 * 2.31 + X4 * 1.06-2.99))).

Depending on the obtained value of P, a forecast is made for the course of gastroduodenal pathology in children:

P≥0.36 - unfavorable;

P <0.36 - favorable.

The invention is illustrated by two clinical examples.

Clinical example 1.

Patient K., 12 years old.

He entered the gastroenterological department with complaints of dull aching pain in the right hypochondrium, lasting up to 15 minutes. Pain occurred both on an empty stomach and immediately after eating. In addition, nausea and bitterness in the mouth periodically worry. From the anamnesis it is known that he considers himself sick since 6 years. Eradication therapy was not carried out.

On the basis of clinical and anamnestic data, ultrasound and endoscopy, the diagnosis is made: Chronic gastroduodenitis (hypertrophic antrum gastritis, bulbitis), exacerbation period. Chronic cholecystitis, period of exacerbation.

Blood plasma IgM and S. aureus IgM levels were 2.15 and 6.76 μg / ml, respectively.

Against the background of the therapy (almagel, motilium, vitamins, choleretic, physiotherapy), positive dynamics were noted - by the time of discharge the pain abdominal syndrome was stopped.

We substitute the obtained results in the formula: P = 1 / (1 + exp (- (6.76 * 0.05-2.15 * 0.08 + 1 * 2.31 + 0 * 1.06-2.99)) )

According to our calculations, according to the formula P, this patient was 0.37. This means that this patient can be assigned to a risk group for the development of erosion and ulcers in the stomach and duodenum.

After 2.5 years due to the resumption of complaints, the patient is re-hospitalized. When conducting FEGDS, erosion and a peptic ulcer in the duodenal mucosa are detected in the patient.

Clinical example 2.

Patient G., 15 years old.

Received with complaints of acute pain, mainly in the epigastrium, lasting up to 15 minutes, not related to eating. Pain is provoked by dietary errors and emotional overstrain. In addition, they are concerned: belching rotten, heartburn, bloating, a tendency to constipation. According to the patient, the patient considers himself for two years. Two years before this hospitalization, endoscopic examination revealed hypertrophic gastroduodenitis. H. pylori did not receive eradication.

Objectively, the patient has a normosthenic physique, there are periorbital “shadows”. On palpation - moderate pain in the epigastrium, right hypochondrium and paraumbilical region, the symptoms of Kehr and Ortner are positive. On the basis of clinical and anamnestic data, ultrasound and endoscopy, the diagnosis is made:

Diagnosis: peptic ulcer of the duodenal bulb, period of exacerbation, stage 2, uncomplicated. Chronic gastroduodenitis (superficial pangastritis, bulbitis), period of exacerbation. Chronic cholecystitis, period of exacerbation.

Plasma IgM and S.aureus IgM levels in blood plasma were 35.91 and 24.86 μg / ml, respectively.

Eradication therapy was prescribed, against the background of which there was a positive clinical and endoscopic dynamics: abdominal pain syndrome stopped, again with minor endoscopy, slight superficial gastroduodenitis, DGR.

We substitute the obtained results in the formula: P = 1 / (1 + exp (- (24.86 * 0.05-35.91 * 0.08 + 0 * 2.31 + 0 * 1.06-2.99)) )

According to our calculations, according to the forecast formula, P in this patient was 0.023, i.e. in this patient, we can expect a favorable course of the disease, despite the detection of a ulcerative defect.

As part of the follow-up, after 8 months, the girl undergoes a follow-up examination, during which with FEGDS a limited superficial gastroduodenitis is detected.

The effectiveness of the method is confirmed by dynamic observation for 1-5 years for 53 children with gastroduodenal pathology.

Thus, the proposed method allows with high accuracy (89%) to predict the occurrence and recurrence of erosive and ulcerative lesions of the gastric mucosa and duodenum in children with gastroduodenal pathology, has sufficient sensitivity (84%) and specificity (79%). Timely corrective treatment in children with a high risk of erosion and ulcers in the gastric mucosa and duodenum will significantly reduce the likelihood of complications and improve the prognosis of the disease. The complex of laboratory and anamnestic parameters chosen by us is the most optimal from the standpoint of non-invasiveness and atraumaticity.

Claims (1)

A method for predicting the course of gastroduodenal pathology by collecting venous blood in children with chronic diseases of the stomach and duodenum, characterized in that the plasma obtained by centrifugation determines the level of Ig M to endotoxin and S.aureus using standard enzyme immunoassay, moreover, as an endotoxin antigen use the glycolipid Salmonella minnesota RE 595, S.aureus - ultrasonic disintegrate of these bacteria, determine the number of H. pylori eradications performed and the patient’s gender, in the subsequent calculation There is a predictive probability according to the formula:
P = 1 / (1 + exp (- (X1 * 0.05-X2 * 0.08 + X3 * 2.31 + X4 * 1.06-2.99))),
where P is the prognostic level of probability; predictors: X1 - level of Ig M to S.aureus (μg / ml); X2 - level of Ig M to glycolipid (μg / ml); X3 - floor (w-0, m-1); X4 is the number of Helicobacter pylori eradications performed, with a probability level of P≥0.36, an unfavorable outcome is predicted (a high risk of erosion and ulcers and relapse in the gastric mucosa and duodenal ulcer) and favorable, with P <0.36.