RU2325845C1 - Helicobacter pylori infection non-invasive disease diagnosis - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to the field of medicine, in particular to the non-invasive diseases diagnosis and it can be used for early diagnosis of Helicobacter pylori infection in the gastrointestinal tract. The method provides early detection of Helicobacter pylori infection, and as a result up-to-date diagnosis of erythematic gastroduodenitis, nodularis gastritis, gastric erosion, onion duodenum ulcer, reflux esophagitis, duodenogastric reflux, overall time of analysis reducing, significant increasing of the reliability of the results, simplification in its implementation, and the reduction of discomfort while diagnosing the upper gastrointestinal tract, including the examination of toddlers and children of one year. The patient should intake an aqueous solution of urea concentration 16,7 mg /ml 5 min prior to the examination, followed by the selection of air samples from the mouth, injection in the test device, the analytical signal recording As a test device for the ammonia registration you should use the closed cell detection with one piezoelectric temperature transducer, which electrodes have the selective film of the acid For this reason the sallcyl-sulphonic acid aqueous solution should be prepared which is then placed on the piezoelectric temperature transducer electrodes so that the mass of the film after solvent removing would be 50-60 micrograms The piezoelectric temperature transducer with sallcyl-sulphonic acid should be dried in the drying oven, and then it should be cooled and joined to the cell detection cover Then the test device is used for infection detection in the gastrointestinal tract in three phases Before infection activating by the carbamide the basic level of ammonia content in the exhaled air should be identified For this reason the exhaled air of 3-5 m is injected into the cell, frequency oscillations of the piezoelectric temperature transducer with sallcyl-sulphonic acid are recorded until the permanent value, changing by less than 5 Hz (F_b) After carbamide activating the ammonia concentration load level is evaluated according to the signal of the piezoelectric temperature transducer with sallcyl-sulphonic acid (F load) The conclusion on the presence and content of the Helicobacter pylori bacteria in the gastrointestinal tract is made on the signal difference of the piezoelectric temperature transducer with sallcyl-sulphonic acid before and after the patient intakes the carbamide ΔF=F_load-F_b and nomogram The test is considered to be positive for the presence of bacteria if the load is more than basic level of ammonia by 250-300 Hz, and if the value difference is greater than 650 Hz the diagnosis is performed on the presence of amines in exhaled air, indicating a profound defeat of the gastrointestinal tract.

EFFECT: providing the early Helicobacter pylori infection detection and as a result up-to-date diagnosis of erythematic gastroduodenitis, nodularis gastritis, gastric erosion, onion duodenum ulcer, reflux esophagitis, duodenogastric reflux; decrease of diagnosis time and increase of result reliability

1 tbl, 1 dwg, 1 ex

Description

The invention relates to non-invasive diagnosis of diseases and can be used for early diagnosis of Helicobacter pylori gastrointestinal tract infection, monitoring a patient during anti-Helicobacter therapy and monitoring its effectiveness.

The closest in technical essence and the achieved effect is a method for determining Helicobacter pylori using indicator test tubes [A.V. Budanov, A.V. Golikov, A.G. Elepov, A.V. Kozlov // "Medical equipment and medical devices ". - No. 3 (14) - June, 2003].

The disadvantage of the existing method is the subjectivity of determining the concentrations, the difficulty of interpreting the results, the need for preliminary aspiration of exhaled air (within 10 minutes), the inability to reuse the device even when the filler is updated, the lack of software support for decision making, and the interfering effect on the results of the analysis of amines and water vapor .

An object of the invention is to develop a method for early detection of Helicobacter pylori infection, simple to implement, which allows to reduce the morbidity and discomfort of the examination of the upper gastrointestinal tract, including examination of children of one year old and toddler, which allows timely diagnosis of erythematous gastroduodenitis, nodular gastritis, erosion stomach ulcer, duodenal bulb, reflux esophagitis, duodenogastric reflux, reduce the total analysis time, significantly to increase the reliability of the results obtained, to simplify the stage of sampling the air exhaled by the patient.

The stated technical problem is achieved by the fact that in the method of non-invasive diagnosis of Helicobacter pylori infection, which involves the activation of bacteria in the gastrointestinal tract by taking a patient 5 minutes before examining an aqueous solution of urea with a concentration of 16.7 mg / ml, taking a sample of exhaled air, injecting a test device, recording an analytical signal, assessing the load level of ammonia concentration and deciding on the activity of infection, new is that as a test device for recording ammonia is used a closed detection cell with one piezoelectric crystal is used, the electrodes of which are modified with a highly sensitive and selective sulfosalicylic acid film (sensor), for which an aqueous solution of sulfosalicylic acid with a concentration of 10 μg / μl is prepared, which is then applied to the electrodes of the piezoelectric crystal so that the film mass after removal the solvent by drying in an oven was 50-60 μg, the finished sensor is cooled and attached to the cover of the detection cell, and then carry out bsledovanie patients for the presence of infection in the gastrointestinal tract in three phases: before activation urea infection estimate baseline ammonia in exhaled air, which are injected into the cell of 3-5 cm ³ exhaled air, fixed change in the oscillation frequency of the sensor to a constant value F _b after activation urea estimate load levels of ammonia concentration sensor signal F on _heating and the conclusion about the presence and the contents in the gastro-intestinal tract of Helicobacter pylori bacteria make the difference signal Sens ora before and after the patient has taken urea (ΔF = F _heat- F _b ) and a nomogram that allows the bacterial content to be determined to be “insignificant”, “does not exceed the norm”, “exceeds the norm”, “significantly exceeds the norm”.

The technical result of the invention is to expand the capabilities of non-invasive methods for diagnosing the presence of Helicobacter pylori microorganisms in the human gastrointestinal tract, reducing the total analysis time, significantly increasing the selectivity and reducing the determination error.

A non-invasive method for diagnosing Helicobacter pylori infection is as follows.

An aqueous solution of sulfosalicylic acid with a concentration of 10 μg / μl is prepared, then the sorbent solution is applied to the electrodes of the piezoelectric crystal so that the film mass after removal of the solvent is 50-60 μg. The finished sensor is cooled and attached to the cover of the detection cell, after which patients are examined for the presence of infection in the gastrointestinal tract.

The methodology for examining patients includes several stages:

1. Preparation of the test system: a hermetically sealed test device (ammonia vapor sensor) is opened, placed in a closed detection cell, connected to a frequency meter connected to a personal computer (or without it);

2. Assessment of the baseline: an air sample is taken from the patient’s mouth with a medical syringe without a 3-5 cm ³ needle, avoiding saliva, inject it into the detection cell and record the frequency counter until a constant (± 5 Hz) value of F _b , Hz is established.

3. Activation of the bacteria: the patient takes a urea solution 5 minutes before the start of the examination to increase the concentration of ammonia in the exhaled air, adequate to the content of the bacterium Helicobacter pylori (Hb);

4. Assessment of the load level: air is taken from the patient’s mouth again with a medical syringe without a 5 cm ³ needle, avoiding saliva, inject it into the detection cell, and constant (± 5 Hz) readings of the frequency meter F _heats , Hz are recorded.

определяют концентрацию аммиака в выдыхаемом пациентом воздухе и содержание бактерии Hb в желудочно-кишечном тракте по номограмме (см. чертеж). Оценивают содержание бактерии с оценками «незначительное», «не превышает норму», «превышает норму», «значительно превышает норму».5. Processing the analysis results: calculate the difference in the readings of the frequency meter before and after taking the urea solution ΔF = F _load -G _b , according to the calibration graph

determine the concentration of ammonia in the air exhaled by the patient and the content of the Hb bacteria in the gastrointestinal tract according to the nomogram (see drawing). The bacterial content is assessed with the ratings “insignificant”, “does not exceed the norm”, “exceeds the norm”, “significantly exceeds the norm”.

The positive effect of the proposed method is achieved by hydrolysis of the urea substrate with the enzymes of the bacterium Helicobacter pylori and the release of gaseous ammonia during the reaction. The ammonia concentration is recorded by a highly sensitive and selective test device based on a piezoelectric crystal transducer modified with a sulfosalicylic acid film (sensor).

The drawing shows a diagram for determining the content of the Helicobacter pylori bacteria in the patient's body and the concentration of ammonia in the exhaled air.

The method is illustrated by the following example.

Example

A method for non-invasive diagnosis of Helicobacter pylori infection consists of several stages.

1. An aqueous solution of sulfosalicylic acid with a concentration of 10 μg / μl is prepared, which is then applied to the electrodes of the piezoelectric quartz AT-section transducer (natural vibration frequency of 8-10 MHz) using a microsyringe so that the film mass after removal of the solvent in an oven is 50- 60 mcg. The finished sensor is cooled and connected to the cover of the detection cell with a closed input, with access to the signal recording unit (frequency counter, personal computer), the signal stability is controlled (± 5 Hz) and packaged in a sealed enclosure.

2. Preparation of the test system: a hermetically sealed test device (ammonia vapor sensor) is opened, placed in a closed detection cell, connected to a frequency meter connected to a personal computer (or without it);

3. Assessment of the baseline: an air sample is taken from the patient’s mouth with a medical syringe without a 5 cm ³ needle, avoiding saliva, inject it into the detection cell and record the constant readings of the frequency meter F _b , Hz, changing by no more than 5 Hz.

4. Activation of the bacteria: the patient takes a urea solution with a concentration of 16.7 mg / ml 5 minutes before the measurement to increase the concentration of ammonia in the exhaled air, adequate to the content of the bacterium Helicobacter pylori (Hb);

5. Assessment of the loading level: 5 minutes after taking the urea solution, air is again taken from the patient’s mouth with a medical syringe without a 5 cm ³ needle, avoiding saliva, inject it into the detection cell and record the readings of the frequency meter F _heated. , Hz, changing by no more than 5 Hz.

определяют концентрацию аммиака в выдыхаемом пациентом воздухе и пропорциональное ей содержание бактерии Hb в желудочно-кишечном тракте по номограмме (см. чертеж). Оценивают содержание бактерии как: «незначительное», «не превышает норму», «превышает норму», «значительно превышает норму».Processing the results of the analysis: calculate the difference in the readings of the frequency meter before and after taking the urea solution ΔF = F _load -F _b , according to the calibration graph

determine the concentration of ammonia in the air exhaled by the patient and the proportion of Hb bacteria in the gastrointestinal tract proportional to it by the nomogram (see drawing). The bacteria content is estimated as: “insignificant”, “does not exceed the norm”, “exceeds the norm”, “significantly exceeds the norm”.

The test is considered positive (HP +) if the load level of the piezosensor of ammonia vapor (ΔF) after taking urea is more than the base (initial) by 250-300 Hz (critical level):

ΔF = F _b -F _load ≥300 Hz,

where F _b is the piezosensor signal during sorption of exhaled air before receiving a urea solution, MHz (background level assessment); F _load - a piezosensor signal during sorption of exhaled air after receiving a urea solution, MHz; ΔF is the difference of the signals of the piezosensor before and after receiving carbamide, Hz.

If the difference in sensor signals before and after taking carbamide is more than 650 Hz, then in the expired air amines are present, indicating a deep lesion of the gastrointestinal tract.

When implementing the method of non-invasive diagnosis of Helicobacter pylori infection, the general requirements for preparing the patient for an examination that is carried out on an empty stomach, at least three hours after eating, are preserved.

Before testing, the patient should not consume or take:

- antibiotics and antisecretory drugs two weeks before the examination,

- antacid preparations, dosage forms containing anti-inflammatory drugs 2-3 days before the examination (some forms of analgesics can inhibit the activity of Helicobacter pylori),

- spirits for three days before the examination,

- legumes during the day before the examination,

- chewing gum containing urea 3 hours before the examination,

- it is recommended not to smoke 2-3 hours before the start of the examination,

- It is recommended to brush your teeth and rinse your mouth thoroughly before testing.

The method is feasible.

Comparative characteristics of the proposed method and prototype are shown in the table.

Table
Comparative characteristics of test devices (prototype, inventive method) for non-invasive diagnosis of Helicobacter pylori infection Criteria Way Prototype The claimed The range of operation of the device depending on the concentration of ammonia vapor in the gas phase 4-30 mg / m ³ 0.04-10 mg / m ³ Method for recording an analytical signal visual (the appearance of a blue color of the indicator layer) frequency change of a piezoelectric crystal transducer (ΔF, Hz) Air sampling device + + The minimum volume of air sample, cm ³ fifty 3 The need for pre-aspiration + - Regeneration and reusability - - Possibility of reuse when updating the surface of the sensitive layer - + Disturbing effect of components amines, water vapor not found Test Analysis Cost 350 rub. 50 RUR The subjectivity of determining concentrations + - Difficulty interpreting the results + - The dependence of the test results on the physical activity of the patient - - Analytical signal recording time 5 minutes 5-10 s Total analysis time 20-25 min 6 min Decision support software - + Reproducibility of analysis results,% 5-10 2-3 Service life 2 months 12 months The dependence of the shelf life of the device on external factors collapses in direct sunlight, mechanical shock, close contact with ammonia sources loss of sensitivity upon close contact and storage with ammonia sources, fragile

The proposed method for the diagnosis of Helicobacter pylori infection is expressed, which allows to reduce the morbidity and discomfort of examining the upper gastrointestinal tract, including when examining children of preschool and nursery age, to significantly increase the reliability of the results, it is easy to implement, does not require expensive equipment, including special endoscopic, prolonged selection of exhaled air, blood sampling, special training and highly qualified personnel.

From the example, figures and tables it follows that a positive effect by the method of non-invasive diagnosis of Helicobacter pylori infection is achieved by preliminary assessment of the baseline level of ammonia concentration in exhaled air of 3-5 cm ³ volume, subsequent activation of infection (5 minutes before the examination of the aqueous urea solution with a concentration of 16.7 mg / ml) and re-measuring the concentration of ammonia in the breath volume of 5.3 cm ³ using as the test device a piezoelectric transducer, a modified film sul acid osalitsilovoy weighing 50-60 g (sensor). The conclusion about the presence in the gastrointestinal tract and the number of Helicobacter pylori bacteria is made by exceeding the load (critical) level of ammonia in the patient's oral cavity, based on the calibration graph and nomogram for the difference in sensor signals ΔF = F _heat -F _b .

Changing the nature of the modifier of the piezoelectric crystal, its mass, the sequence of analysis, reducing or increasing the volume of exhaled air, or not following the instructions before the test makes it impossible to implement the method due to overload or low sensitivity of the sensor to ammonia, loss of selectivity and reproducibility of signals.

The proposed method for non-invasive diagnosis of Helicobacter pylori infection can reduce the morbidity and discomfort of the examination of the upper gastrointestinal tract, therefore, it can be recommended for examination of children of preschool and nursery age. The method is simple, express, easy to implement, economical, accessible to the general public, does not require special endoscopic and other expensive equipment, a blood sampling procedure, the availability of qualified personnel, and also allows the early detection of Helicobacter pylori infection, timely diagnosis of erythematous gastroduodenitis, nodular gastritis , gastric erosion, duodenal ulcer, esophagitis reflux, duodenogastric reflux, observe the patient during anti-Helicobacter pylori therapy and monitoring its effectiveness.

Claims (1)

A method for non-invasive diagnosis of Helicobacter pylori infection, including taking a patient 5.7 minutes before the start of examination of an aqueous urea solution with a concentration of 16.7 mg / ml, sampling air from the oral cavity, injecting into a test device, registering an analytical signal, characterized in that As a test device for recording ammonia, a closed detection cell with one piezoelectric crystal is used, the electrodes of which are modified with a selective film of sulfosalicylic acid, for which an aqueous solution is prepared sulfosalicylic acid, which is then applied to the electrodes of the piezoelectric crystal so that the film mass after removal of the solvent is 50-60 μg, the finished piezoelectric crystal with the sulfosalicylic acid film is dried in an oven, then cooled and attached to the lid of the detection cell, after which a test device examining patients for infection in the gastrointestinal tract in three stages: before activating the infection with urea, the baseline level of ammonia in expired air, which are injected into the cell of 3-5 cm ³ exhaled air, fixed frequency change film piezoelectric transducer with sulfosalicylic acid oscillations to a constant value that varies by no more than 5 Hz (F _b), after activating the urea concentration estimate load levels ammonia on a signal from the piezoelectric film transducer sulfosalicylic acid (F _heating), and the conclusion about the presence and the contents in the gastro-intestinal tract of Helicobacter pylori bacteria make the difference signals pezokva tsevogo converter with sulfosalicylic acid film before and after the patient urea ΔF = F _b -F _LOAD and nomogram, the test is considered positive for the presence of bacteria if the load level of ammonia greater base at 250-300 Hz, and when it is larger difference values 650 Hz diagnose the presence in the expired air of amines, indicating a deep lesion of the gastrointestinal tract.

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