RU2739222C1 - Method for diagnosing urinary tract infection in infants and young children - Google Patents

Abstract

FIELD: medicine.SUBSTANCE: invention refers to medicine, namely to pediatrics and pediatric nephrology, and discloses a method of diagnosing urinary tract infection in infants and young children. Method includes urine examination for zonulin protein content, intestinal permeability regulator. Substance of the method consists in determining the concentration of zonulin in a morning portion of urine, before prescribing an antibacterial therapy in infants and young children suspected of urinary tract infection. If the urine sample shows a level of zonulin higher than 0.4 ng/ml (p<0.05), the urinary tract infection is documented in the patients.EFFECT: method provides early safe, non-invasive rapid diagnosis of urinary tract infection in infants and young children and enables timely determination of the pathogenetic therapy approach.1 cl, 3 ex

Description

The invention relates to medicine, namely to pediatrics and pediatric nephrology, can be used as an express diagnosis of UTI in infants and young children.

A method for diagnosing urinary tract infection (UTI) in infants and young children is described. The method includes testing urine for the protein content of zonulin (Zo), a regulator of intestinal permeability. Zo has potential value in determining the mechanism of the paracellular “gateway” in the intestine, forming a tight junction (TJ) in the apical part of the intestinal endothelium [1, 2]. The zonulin protein is one of the main factors providing an adequate action of the gut mechanism, reversibly affecting the tightness of the TJ. Experimental studies have shown that an increase in Zo concentration in parallel contributes to an increase in intestinal permeability [1, 2]. As a result of a comprehensive study, a link was proved between increased intestinal permeability and biomarkers of inflammatory and endothelial dysfunctions [3]. When exposed to any damaging agents (bacteria, parasites, viruses, allergens), the integrity of the intestinal barrier is violated, dysregulation occurs in the intestines, autoflora multiplies, which contributes to systemic bacterial inflammation, including renal tissue [1, 2, 3, 4].

There are various methods for diagnosing UTIs [5-8]. A known method for the diagnosis of pyelonephritis in children [9], inviting all children with acute pyelonephritis to investigate the middle portion of morning urine and in aggregate to use standard laboratory tests: polymerase chain reaction (PCR) and the microbiological method. Using PCR, it is recommended to detect the following pathogens: DNA of herpesvirus infections, chlamydia, mycoplasma and human papillomavirus infections. Using the microbiological method - to determine the quantitative and specific characteristics of uropathogens. If the above pathogens are found in the urine, as well as the contamination of urine with bacteria in the amount of 100 thousand CFU / ml, the authors of the method suggest diagnosing chronic pyelonephritis. This method is not informative and specific enough, the diagnostic process can be extended over time (according to the research standards, the bacteriological examination of urine takes up to 7-10 days on average) and often requires differential diagnosis with other non-inflammatory kidney diseases. According to the clinical recommendations of the Union of Pediatricians of Russia, the etiological agent for the development of UTI in children is a bacterial infection with a predominance of gram-negative flora, and viral infection is not among the causes of the development of UTI [5].

A known method for the diagnosis of acute pyelonephritis [10], based on the determination of the activity of alkaline phosphatase (ALP) in the blood leukocytes of a patient with urinary tract infection. The authors of the method recommend determining the average histochemical coefficient of the alkaline phosphatase level before and after incubation of a blood sample with a renal antigen, if the difference in the obtained indicator in comparison with the initial value is from 0.2 to 3.0, acute pyelonephritis is diagnosed. Of the shortcomings of this method, it is worth noting that a number of laboratories may have difficulties in staging reactions, due to the lack of the necessary amount of equipment for staging research. Also, this method is recommended for the diagnosis of pyelonephritis in elderly and senile people and cannot be used in infants and young children.

A known method for the diagnosis of acute pyelonephritis in children by the level of expression of adhesion molecules CD11β and CD54 by peripheral blood lymphocytes [11]. Using the method of indirect immunofluorescence, the percentage of monoclonal antibodies in children with clinical manifestations of pyelonephritis is determined. According to studies, with a CD11β level of more than 26.34% and a CD54 level of more than 33.26%, a urinary tract infection is diagnosed. The disadvantage of this method is that this method can not always be used in the diagnosis of UTI in young children. In most cases, in children under three years of age, there are no signs of a systemic inflammatory reaction [5, 8]. As a rule, clinical symptoms in infants and young patients are mild and do not have the typical clinical symptoms characteristic of older children [5, 8]. Often, the diagnosis of UTI in young children is difficult, and changes in urine tests can be detected accidentally during preventive examinations [5-8]. In addition, it should be noted that the method refers to invasive diagnostic methods and cannot always be performed in infants due to the need to collect a large volume of blood - 5 ml.

Closest to the claimed is a method for the diagnosis of pyelonephritis [12], based on determining the optical density of the test urine before and after incubation with a test microorganism in a thermostat. The content of beta-lysines is determined by the degree of lysis of the hay bacillus test culture. With an increase in beta-lysines in comparison with the norm by two or more times, the latent form of pyelonephritis is diagnosed. The advantages of this method are non-invasiveness, simplicity in execution, it is possible to diagnose latent forms, it is recommended to use it in patients with enuresis. Among the shortcomings - there is no data on the possibility of using the technique in children under 3 years old, it is impossible to diagnose acute forms of the disease.

The objective of the claimed method is to develop a non-invasive method for the rapid diagnosis of UTI, which provides early and effective diagnosis in children of the first three years of life.

The essence of the method is to determine the concentration of zonulin in the morning portion of urine, before the appointment of antibiotic therapy, in infants and young children with suspected UTI. If zonulin levels above 0.4 ng / ml (p <0.05) are detected in urine samples, patients will have a UTI. EFFECT: method provides early safe, non-invasive express diagnostics of UTI in infants and young children and allows timely determination of pathogenetic therapy tactics.

The technical result from the implementation of the proposed method lies in the fact that the claimed method allows you to obtain reliable information on the diagnosis of UTI in infants and young children.

The technical result from the implementation of the proposed method:

- complete safety for the patient's health;

- the possibility of conducting research in infants and young children;

- a small amount of biological material is required for the study (5 ml of urine);

- lack of reactogenicity on the part of the patient's body;

- the collection of material does not present any difficulties for parents, it can be done at home;

- allows you to diagnose the disease at an early stage.

The proposed assessment method is technically simple, non-invasive, informative, its implementation does not require large economic costs. The claimed method for the diagnosis of UTI in infants and young children is new and is not described in the publicly available literature. The technical result is achieved as a result of the implementation of the proposed method, the essence of which lies in the fact that the child's urine is examined with the determination of the content of zonulin protein in it by the method of enzyme-linked immunosorbent assay (ELISA) on an automatic enzyme immunoassay analyzer using reagents from Cloud-Clone Corp (CCC, USA) ... At a Zo level above 0.4 ng / ml, a UTI is recorded in patients. The claimed distinctive features are new and in infants and young children the determination of zonulin in urine has not been previously used. Zonulin is a diagnostic marker of intestinal endothelial damage [1, 4, 13, 15], therefore, an increase in this marker indicates damage to the histohematogenous barrier under the influence of various agents [1, 2, 3, 13, 14, 15]. The use in routine practice of invasive diagnostic methods in infants and young children limits and delays the diagnosis, contributes to the formation of chronic forms of diseases. In this regard, the development and introduction into clinical practice of non-invasive diagnostic methods, timely prediction of the exacerbation of the disease and the conduct of predictive therapy are especially relevant in pediatric practice. The method is carried out as follows. If a patient suspects a UTI, urine is collected in an amount of 5 ml into a sterile tube. The material is taken in the morning, after the toilet of the genitals, before the appointment of antibiotic therapy. The biological material is centrifuged for 20 minutes, after which the material is poured into epindorfs. The determination of Zo is carried out in urine by the enzyme linked immunosorbent assay (ELISA) on an automatic enzyme immunoassay analyzer using reagents from Cloud-Clone Corp (CCC, USA). After 3 hours, the results of the study are evaluated. The obtained values are compared with the standard indicators. At Zo levels above 0.4 ng / ml, patients have a UTI.

The efficiency of the method was proved by clinical and laboratory examination of 44 patients of the Regional Children's Clinical Hospital No. 1 in Vladivostok, aged at the time of participation in the study from 29 days of life to 2 years 11 months and 29 days inclusive, who, as a result of a comprehensive examination, were diagnosed with: urinary tract infection. The diagnosis of the disease was carried out in accordance with the standards of the Federal Clinical Recommendations of the Union of Pediatricians of Russia (2018) [5]. The control group included 20 relatively healthy children, comparable in age and sex, with no history and at the time of the study of kidney disease and changes in urine tests. The groups of examined children did not differ among themselves in terms of age and weight - height indicators (p <0.05). The level of zonulin protein (Zo), a regulator of intestinal permeability, was determined in the urine of all children by the enzyme-linked immunosorbent assay (ELISA) on an automatic enzyme immunoassay analyzer using Cloud-Clone Corp reagents (CCC, USA). The average Zo concentrations in urine in infants and young children with UTIs and in children from the comparison group (control group) were 2.86 ± 0.09 ng / ml and 0.16 ± 0.01 ng / ml, respectively (p < 0.05). Thus, the proposed informative method provides effective, safe and early rapid diagnosis of UTI in children and allows timely determination of the tactics of personalized therapy. Examples:

Clinical example 1. Patient D., 2 years 3 months old, was admitted with complaints of an unpleasant odor of urine, refusal to eat. According to my mother, these complaints have been bothering me for a week. We independently passed a general urinalysis, which revealed the following changes: leukocyturia up to 170 in the field of view, bacteriuria +++. Taking into account complaints and test results, the patient was admitted to the nephrology department. From the anamnesis of life: the child from the first pregnancy, the first birth, grew and developed according to age. She was breastfed for up to a year, complementary foods were introduced by age. Allergic history: food allergy to citrus fruits in the form of rash and itching. Has been registered with a gastroenterologist since birth with constipation syndrome. After a comprehensive examination, the child was diagnosed with UTI. The patient was examined for the level of zonulin in the urine in the following way: before the appointment of antibiotic therapy, in the morning, after using the toilet of the genitals, 5 ml of urine was collected in a sterile test tube, then the biological material was centrifuged for 20 minutes, after which the material was poured into epindorfs. Zo was determined in urine by enzyme linked immunosorbent assay (ELISA) on an automatic enzyme immunoassay analyzer using reagents from Cloud-Clone Corp (CCC, USA). After 3 hours, the results of the study were assessed - the level of zonulin in this patient was 0.76 ng / ml. The test results confirmed the presence of UTI.

Clinical example 2. Patient K. 2 g. 1 month came with complaints of abdominal pain, fever up to 39 ° C, lethargy, decreased appetite. Anamnesis of the disease: during a preventive examination at the age of 1 year, according to the ultrasound data, an expansion of the pelvic-pelvic system of both kidneys to 0.7 mm was noted. Subsequently, she was not examined for this. During the last year, there were episodes of subfebrile condition without catarrhal phenomena. Ill for 4 days when the above complaints appeared. From the anamnesis of life it is known that the girl is from a second pregnancy, second birth. She grew and developed according to age norms. From the anamnesis: the child's mother and maternal grandmother suffer from pyelonephritis. A girl at the age of 1 year 3 months suffered an intestinal infection (received Amoxicillin), after which there was a tendency to retain stool for up to 3-4 days. A comprehensive clinical and diagnostic examination was carried out, as well as determination of the content of zonulin in urine according to the proposed method. The result is the following value: 2.61 ng / ml. According to the invention, the diagnosis was made: UTI. The results of the provocative tests performed confirmed the diagnosis of UTI.

Clinical example 3. Patient C., 1 year old. When passing the planned medical commission, changes in the analysis of urine were revealed in the form of leukocyturia 50-60 in the field of view. From the anamnesis: grew and developed according to age. At 1 month, ultrasound of the kidneys revealed pyelectasis on the right 0.6 mm. Further, kidney ultrasound was not repeated. The child's mother has hydronephrosis on the right, the paternal grandmother suffers from urolithiasis. The patient underwent a comprehensive clinical and laboratory examination and determined the level of zonulin according to the proposed method described in the first example. The result is the following value: 1.2 ng / ml. According to the invention, the diagnosis was made: UTI.

Information sources:

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Claims (1)

A method for diagnosing urinary tract infection in infants and young children, including the collection of a morning portion of urine in a volume of 5 ml, after the toilet of the genitals and before the appointment of antibiotic therapy, characterized in that the content of zonulin protein is determined in the urine, when the concentration of zonulin increases above 0, 4 ng / ml in infants and young children is diagnosed with a urinary tract infection.