Instant diagnostic technique for acute intestinal infections

FIELD: medicine.

SUBSTANCE: invention represents an instant diagnostic technique for acute intestinal infections (AIIs), involving detecting indication markers of the AII aetiology with the use of laboratory immunology tests, differing by the fact that the AII aetiology is stated in children of an early age category, preferentially in the newborn children; that is accompanied by measuring the concentration of cytokine, interleukin IL-10 in coprofiltrate and diagnosing chronic placental insufficiency (CPI); a probability (P) of the bacterial AII aetiology is calculated; the value P of more than 50% testifies to the bacterial AII aetiology, while the value P being less than 50% shows the absence of the bacterial AII aetiology, and enables considering the diagnostics second stage to be necessary, which implies measuring the concentration of cytokine, interleukin IL-4 in coprofiltrate; the time of latching the newborn child to the breast is established with considering the type of feeding; that is combined with calculating a probability (P) of the viral or viral-bacterial AII aetiology, with the value P of more than 50% testifying to the viral AII aetiology, while the value being less than 50% makes it possible to state the viral-bacterial AII aetiology.

EFFECT: more accurate diagnosing of the aetiology of acute intestinal infection and simplifying the diagnostic procedure.

2 tbl

 

The invention relates to the field of medical diagnostics, namely the identification of the etiology of acute intestinal infections, preferably in children of early age, especially in infants.

From acute diarrhea die up to 5 million children per year. The incidence of adults and children OKA in the Russian Federation is also high. In the structure of the gastrointestinal tract in newborn one of the leading syndrome dyspepsia. According to several authors, the structure of the syndrome dyspepsia is dominated by OKA various etiologies (Anokhin Century A. and others, 2006; cottage of wattle A. N. and others, 2005, 2006). OKA occupy a leading place in the structure of infectious diseases of childhood, being one of the main causes of mortality. Maximum threat intestinal infections pose to children of early age, especially for newborns.

Etiological explanation OKA on clinical data in sporadic cases. A significant role in the etiology of intestinal infections in children belongs bacterial agents (Tikhomirov, O. C. et al., 2006). However, in recent years there is increasing evidence of significant role of conditionally pathogenic microorganisms, viruses, fungi in the etiology of acute intestinal infections in children. While the reliability of the etiological decryption of acute intestinal infections remains low the th, what hampers the etiotropic therapy.

To confirm the diagnosis using bacteriological methods: cultures of faeces, blood, vomit, and serological methods for detection in the dynamics of the level of specific antibodies to the bacteria (see Infectious diseases in children. Edited by C. C. Ivanova, Izd-vo: Medical information Agency, 2002, S. 924).

The disadvantage of these methods is the low efficiency and complexity.

There is a method of identifying bacterial infections, comprising the reaction of indirect haemagglutination (rnga), including the preparation and study of serum using specialized diagnostic, including incubation study and control of materials at room temperature for 14-18 h, with subsequent evaluation of the results (see EN No. 2329507, G01N 33/577, 2008). This method allows to obtain good results. However, it takes a long time (24 h), time-consuming and to obtain the results required additional equipment (microplates and spectrophotometer), drugs cannot long keep and take pictures. In some cases the definition of serological markers ineffective: in newborns due to the Mature immune system, depression of the immune system and in the early stages of the disease, because antibodies are formed otsrochennoe

There is also known a method of rapid diagnosis of acute intestinal infections, including identifying marker indicators etiology OKA, using immunological laboratory tests (see EN No. 2329507, G01N 33/577, 2008). The method is characterized by the fact that carry out the detection of antigens of pathogens OKA, which explore lymphocytic suspension (requires at least 0.05 ml lymphocytic mist) for smear preparation, conduct immunocytochemically painting and adequate evaluation. As of a detection system using polymer complex Super Sensitiv Polimer-HRP Detection System production BioGenex, as more sensitive.

Blood is a complex and sensitive subject when conducting immunohistochemically research. In this regard, the proposed new method for the diagnosis of OKA, based on the detection of antigens of pathogens in blood cells requires a complex comprising 16 methodological techniques related to blood collection, centrifugation and deposition on a glass slide (smear), drying at room temperature, fixation in pure acetone, repeated washing in phosphate-buffered saline, treatment in 3% H2O2, incubation in normal blocking serum (Power Block), drawing on strokes poly - or monoclonal antibodies to target antigens, cu the purpose of these antibodies at 37°C, rinse in water and a new washing in phosphate-buffered saline, causing strokes on Super Enhacer, incubation in the dark chamber at room temperature, a new washing in phosphate-buffered saline, drawing on strokes SS Label, incubation in a dark chamber at room temperature, the processing material in an aqueous solution containing 0.05% 3,3-diaminobenzidine tetrachloride (DAB) and 0.025% hydrogen peroxide, weak zakreski with hematoxylin, microscopy.

The disadvantage of this method is its focus on the identification of the etiology of bacterial diseases (dysentery, salmonellosis, typhoid fever, E. coli infection and others) and the inability to detect viral or viral-bacterial etiology.

The objective of the proposed method is the possibility of operational detection of both bacterial and viral or viral-bacterial etiology of acute intestinal infections.

The technical result obtained by the solution of the problem, which seeks to ensure the possibility of rapid detection of both bacterial and viral or viral-bacterial etiology of acute intestinal infections. In addition, simplifies the process of diagnosing.

To solve this problem the method of rapid diagnosis of acute intestinal infections (AII), including the identification marker indicators etiology OKA, using the m immunological laboratory tests, differs in that the etiology of the OKA set of children of early age, preferably in newborns, when this concentration is determined in coprofilia cytokine - interleukin IL-10 and the presence of chronic placental insufficiency (hfpn), and then calculate the probability of bacterial etiology OKA using the expression:

P=(ey/(l+ey))*100%,

where P is the probability in % bacterial etiology OKA;

y=11,4796-5,6647·X1-1,251912·X2;

X1- the presence or absence of hfpn, respectively 1 or 0;

X2- concentration in coprofilia cytokine - interleukin IL-10, PG/ml, with a P value greater than 50% indicates a bacterial etiology OKA, and less than 50% indicates the absence of bacterial etiology of the OKA, and the need for a second stage of diagnosis, which determine the concentration in coprofilia cytokine - interleukin IL-4, identify the period of breastfeeding and infant feeding, they expect the probability of viral or viral-bacterial etiology of the OKA, which use the expression:

P=(ey/(1+ey))*100%,

where P is the probability in % viral or viral-bacterial etiology OKA;

y=-43,95582+2,05532·X3-4,11453·X4+7,01455·X5;

X3through what period of time from the birth of prikladyvat and chest h;

X4- type of feeding (or natural, or mixed or artificial), respectively, or 1, or 2, or 3;

X5- concentration in coprofilia cytokine - interleukin IL-4, PG/ml, with a P value greater than 50% indicates a viral etiology of the OKA, and less than 50% indicates viral-bacterial OKA.

Comparative analysis of the characteristics of the claimed solution with the characteristics of the prototype and analogues demonstrates compliance of the claimed solution to the criterion "novelty".

Signs of a distinctive part of the formula of the invention provide a solution to complex functional tasks.

Signs of "etiology OKA set of children of early age, preferably in newborns allow you to diagnose most children of early ages.

Signs indicating that as the material for research use coprophiliac provide noninvasive sampling of biological material, in addition, select the material most intensive contact with pathogenic microflora (e.g., compared to urine or blood).

Signs indicating that "concentration is determined in coprofilia cytokine - interleukin IL-10", provide the ability to determine the anti-inflammatory cytokine that reflect the immune status of the organism in infectio the different processes, and thereby receive one of the parameters, allowing to carry out the mathematical probability score OKA bacterial etiology.

Signs that identify the presence of chronic placental insufficiency (hfpn)" allow a method for the clinical survey (without laboratory tests at this stage) to obtain a second parameter that allows you to perform mathematical probability score OKA bacterial etiology.

The signs indicate that, after identifying the called indicator indicators calculate the probability of bacterial etiology using the expression:

P=(ey/(1+ey))*100%,

where P is the probability in % of the bacterial etiology of the disease;

y=11,4796-5,6647·X1-1,251912·X2;

X1- the presence or absence of hfpn, respectively, 1 or 0;

X2- concentration in coprofilia cytokine - interleukin IL-10, PG/ml", provide the estimated generating a diagnostic test to detect the presence or absence of bacterial etiology OKA.

Signs indicate that the P value is greater than 50% indicates a bacterial etiology of the disease, and less than 50% indicates the absence of bacterial etiology of the disease and the need for the second stage of diagnosis are diagnostic criterion is the presence or absence of b is serialno etiology OKA.

Signs indicating that "concentration is determined in coprofilia cytokine - interleukin IL-4", provide the ability to determine the anti-inflammatory cytokine that has anti-inflammatory effect in infectious processes, and thereby receive one of the parameters that allow for mathematical assessment of the probability of disease of viral or viral-bacterial etiology.

Signs that reveal the period of breastfeeding", allow a method for the clinical survey (without laboratory studies at this stage) to get another option to spend a mathematical assessment of the probability of viral or viral-bacterial etiology OKA.

Signs that reveal "infant feeding", allow a method for the clinical survey (without laboratory tests at this stage) to get one of the parameters that allow for mathematical assessment of the probability of viral or viral-bacterial etiology OKA.

Signs indicating that "calculate the probability of viral or viral-bacterial etiology, why use the expression:

P=(ey/(1+ey))*100%,

where P is the probability in % viral or viral-bacterial etiology OKA;

y=-43,95582+2,05532·X3-4,11453·X4+7,01455·X5;

X3through what period of hatching of the child were put to the breast, h;

X4- view breastfeeding or natural, or mixed, or artificial, respectively, or 1, or 2, or 3;

X5- concentration in coprofilia cytokine - interleukin IL-4, PG/ml", provide the estimated generating a diagnostic test to detect the presence of viral or viral-bacterial etiology OKA.

Signs indicate that the P value is greater than 50% indicates a viral etiology of the disease, and less than 50% indicates viral and bacterial etiology of the disease are the diagnostic criteria for the presence of viral or viral-bacterial etiology OKA.

Coprophiliac prepared in a known manner: a sample of stool is placed in pre-weighed sterile, with a capacity of 10 ml test tube, then 100 mg of feces was intensively shaken with 5.0 ml of physiological solution (NaCl and 0.09%) solution before the formation of a homogeneous suspension or homogenized samples on mnogoobraznom vortex vigorously Shakira (highest speed) for 30 min, then the homogenate was transferred into 2 ml microcentrifuge tube and centrifuged 5 min at microcentrifuge with the speed of 3000 rpm, the supernatant was collected in a clean labeled tube.

The concentration of cytokines in coprofilia: interleukin IL-10, IL-4 was determined in a known manner, by enzyme-linked immunosorbent Ana is iza using reagents R& D Diagnostics Inc. (USA), in accordance with the instructions on the analyzer Multiscan (Finland). The calculations of a number of cytokines such as IL-10, IL-4 was performed by constructing a calibration curve using a computer program. The number expressed in pilgrammage per milliliter (PG/ml). Analysis of the levels of cytokines carried out in the shortest possible time after the onset of the disease (in fact, immediately after delivery of the child in the clinic).

In addition to laboratory studies clinical survey or study of the medical records of the child to detect the presence or absence of chronic placental insufficiency, the presence or absence of hfpn corresponds to a value of 1 or 0 index X1.

Next, after determining the index of X2- concentration in coprofilia cytokine - interleukin IL-10, PG/ml on the first of the above formula calculates the probability of bacterial etiology.

If the calculation shows that the P value is greater than 50%, it indicates bacterial etiology of the disease, if R is less than 50%, it indicates the absence of bacterial etiology of the disease and the need for the second stage of diagnosis of the etiology of the disease, to install it viral or viral-bacterial etiology.

In laboratory studies it is advisable you who manage a number of both cytokines (IL-4 and IL-10), since the length of the analysis in their ratio is 4 hours, so you can save time when not acknowledge assumptions about the bacterial etiology of the disease, in addition, for the same reason, it is advisable at this stage to set the indicator X3- information about how long the period from the birth of the child were put to the breast (measured in hours) and the indicator of X4- information about the form of feeding, which may be either natural, or mixed, or artificial (which correspond to the values of the metric, PG/ml);

Next, after determining the index of X5- concentration in coprofilia cytokine - interleukin IL-4, in PG/ml on the second of the above formula calculates the probability of viral or viral-bacterial etiology of the disease, if the P value is greater than 50%, this suggests a viral etiology of the disease, if the P value is less than 50%, this indicates viral and bacterial etiology of the disease.

Examples of detection of bacterial etiology OKA shown in table.1.

Table 1
The detection results of the OKA bacterial etiology
No.Hfpn IL10The etiologyyP
7,09790899,92%
205,64,468892898,87%
313,61,308016878,72%
41a 3.90,932443271,76%
5038,8non-bacterial-37,0945860,00%
6110,2-6,95460240,10%
70of 17.5-10,428860,00%
8 17,9-4,07520481,67%
9010,2-1,289902421,59%
10026,5-21,6960680,00%
11017,9-10,9296250,00%
12012,34-3,96899411,85%

The forecast diagnoses in relation to bacterial etiology, which coincided with actual - 125 of 128, (97,3%).

Some examples of definitions of viral diseases and viral and bacterial etiologies are listed in table.2.

17,8
Table 2
The result of the detection of the OKA viral and viral-bacterial etiology
No.Put to the breast after the hourInfant feeding IL4The etiologyP
10,253104,4viral676,5334100,00%
20,25311,424,18029100,00%
34317,876,78086100,00%
4243314,07192100,00%
50,25315,150,13413100,00%
60,25311,424,18029100,00%
7025 317,869,07341100,00%
80,253104,4676,5334100,00%
90,25311,424,18029100,00%
104317,876,78086100,00%
11243314,07192100,00%
120,25315,150,13413100,00%
130,25311,424,18029100,00%
140,25369,07341100,00%
154317,876,78086100,00%
16243314,07192100,00%
170,25315,150,13413100,00%
180,25311,424,18029100,00%
190,25317,869,07341100,00%
200,253104,4676,5334100,00%
210,2532,7 -36,84630,00%
220,2532,7-36,84630,00%
231232,7-12,69630,00%
240,2522,7-32,73180,00%
250,2532,7-36,84630,00%
261232,7-12,69630,00%
270,2532,7-36,84630,00%
280,2532,7-36,8463 0,00%
290,2532,7-36,84630,00%
300,2522,7-32,73180,00%
311232,7-12,69630,00%
320,2532,7-36,84630,00%
330,2522,7-32,73180,00%
340,2532,7-36,84630,00%

According to the data table of the 51 cases in 100% of cases forecast data coincided with the actual.

Realistically estimated and accounting operations of the method are implemented programmatically, using About the scheme, representing standard file office Suite MicrosoftExcel 2003.

The program allows you to store data on the results of the forecast of diagnosis up to 30 people. Save, copy and reproduction file is provided by standard tools MicrosoftExcel 2003 and Window's 98.

File consists of 2 tabs, "I stage. Bacterial and stage II. Viral".

At the time of admission, the user opens the file tab, "I stage. Bacterial". Filled in:

1) In the column "name of patient" enter the surname, name and patronymic of the patient. This field is not mandatory;

2) In column hfpn (chronic fetoplacental insufficiency) are binary data: 1 -, 0 - no.

3) the column "IL 10 is populated with the results of laboratory research, field enter the appropriate value.

Next, in columns "Forecast of diseases of bacterial etiology, in %" are automatically calculated in percentage probability values, respectively, of the presence of the patient's diseases of bacterial etiology

If the value in column "Forecast of diseases of bacterial etiology,% more than 50%, it is considered that the patient has a bacterial form of the disease and further investigation is not required.

If the value in column "predictors of disease bacterial etiology and, percentage less than 50%, it is deemed that the patient's disease, non-bacterial, and further investigation is required. To do this, the user navigates to the tab "stage II. Viral". Filled in:

1) In the column "name of patient" enter the surname, name and patronymic of the patient. This field is not mandatory;

2) In the column "Duration of breastfeeding through ... hours" enter the number of hours from 1 to 24;

3) In column "Feeding" entered the following values: 1 - natural 2 - mixed, 3 - artificial;

4) In the column "IL 4" enter the value corresponding to the value of this parameter.

Next, in columns "predictors of disease of viral etiology, in %" and "Forecast of diseases of viral and bacterial etiology, in %" are automatically calculated in percentage probability values, respectively, of the presence of the patient's disease of viral etiology or viral and bacterial etiology.

If the value in column "predictors of disease of viral etiology, in %" more than 50%, it is considered that the patient has a viral form of the disease and further investigation is not required.

If the value in column "predictors of disease of viral etiology, in %" less than 50%, it is considered that the patient has viral-bacterial form of the disease.

Further the surveys are not required.

The method of rapid diagnosis of acute intestinal infections (AII), including the identification marker indicators etiology OKA, using immunological laboratory tests, characterized in that the etiology of the OKA set of children of early age, preferably in newborns, when this concentration is determined in coprofilia cytokine - interleukin IL-10 and the presence of chronic placental insufficiency (hfpn), and then calculate the probability of bacterial etiology OKA using expressions:
P=(ey/(1+ey))*100%,
where P is the probability in % bacterial etiology OKA;
y=11,4796-5,6647·X1-1,251912·X2;
X1- the presence or absence of hfpn, respectively 1 or 0;
X2- concentration in coprofilia cytokine - interleukin IL-10, PG/ml, with a P value greater than 50% indicates a bacterial etiology OKA, and less than 50% indicates the absence of bacterial etiology of the OKA, and the need for a second stage of diagnosis, which determine the concentration in coprofilia cytokine - interleukin IL-4, identify the period of breastfeeding and infant feeding, they expect the probability of viral or viral-bacterial etiology of the OKA, which use the expression:
P=(ey/(1+ey))*100%,
where R - Vero is tnost % viral or viral-bacterial etiology OKA;
y=-43,95582+2,05532·X3-4,11453·X4+7,01455·X5;
X3through what period of time from the birth of the child were put to the breast, h;
X4- type of feeding (or natural, or mixed, or artificial), respectively, or 1, or 2, or 3;
X5- concentration in coprofilia cytokine - interleukin IL-4, PG/ml, with a P value greater than 50% indicates a viral etiology of the OKA, and less than 50% indicates viral-bacterial OKA.



 

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1 cl, 3 ex, 1 tbl

FIELD: medicine.

SUBSTANCE: invention refers to medicine, particularly to laboratory diagnostics. Substance of the method: cytomegalovirus antibody titre is measured; disturbed haemoglobin oxygenation in erythrocytes is detected; a histochemical method is used to determine the activity of glucoso-6-phosphate dehydrogenase and the content of erythrocyte methaemoglobin; if the cytomegalovirus antibody titre is 1:1,600, the activity of glucoso-6-phosphate dehydrogenase diseases to 15.0±0.8 standard units, and the content of erythrocyte methaemoglobin increases up to 1.7±0.09% and peripheral blood oxyhaemoglobin decreases up to 90.0±1.3%, threatening hemic hypoxia is stated.

EFFECT: invention can be used to state threatening hemic hypoxia in the pregnant women suffering aggravated cytomegalovirak infection in the third trimester of gestation.

2 dwg

FIELD: medicine.

SUBSTANCE: analysed blood sample is taken that is combined with a dynamic cytofluorometrical analysis of the taken test sample for endothelial dysfunction markers - soluble adhesion molecules sICAM-1 and sVCAM-1. If the expression level of the adhesion molecules sICAM-1 is 591.9 ng/ml and more, and the expression level of sVCAM-1 is 632 ng/ml and more on the first days of a peracute period of the ischemic stroke to be persistent in dynamics or increasing in relation to the reference, the unfavourable course of ischemic stroke in the given category of patients is predicted.

EFFECT: using the method provides high sensitivity, specificity and accuracy of the prediction on the first days following the peracute period of the ischemic stroke in diabetic patients.

5 dwg, 1 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to pathologic anatomy, and enables providing the morphological differential diagnosis of trophoblastic, endometrial and mixed forms of primary placental insufficiency in spontaneous miscarriage in the first trimester. The technique consists in performing an immunohistological analysis of the villous chorion and decidual cells of the gravidary endometrium to evaluate a vascular endothelial expression index (VEEI) and the transforming growth factor (TGF-β2). If the VEEI villous chorion expression index is 1.5 standard units or less, while the TGF-β2 expression index is 1.4 standard units or less, the trophoblastic form of primary placental insufficiency is diagnosed. If the VEEI decidual cell expression index is 1.6 standard units or less, while TGF-β2 is 2.7 standard units or more, the endometrial form of primary placental insufficiency is diagnosed; if observing the above values simultaneously, the mixed form of primary placental insufficiency is diagnosed.

EFFECT: presented method is high-informative, increases the quality of a pathology report, promotes an individual approach to the further diagnosing, treatment and rehabilitation of the female reproductive health depending on the pathogenetic form of placental insufficiency.

3 ex

FIELD: medicine.

SUBSTANCE: invention relates to the field of medicine, in particular to neurosurgery, and deals with a method of diagnosing the brain tumour in patients. The essence of the method: the percent quantity of CD25-lymphocytes is determined in sampled blood. If its value is 13.7 the absence of the brain tumour is stated, in case if its value is lower than 13.7 the presence of the brain tumour in the patient is diagnosed. The method is simple for the practical application.

EFFECT: invention ensures a zero level of errors, which testifies to a high probability of a correct distribution of the tested people for groups of healthy people and patients with the brain tumour, which makes it possible to practically apply the claimed method in the additional diagnostics of a cerebral pathology.

2 tbl, 1 dwg

FIELD: medicine.

SUBSTANCE: pathogenetic treatment of chronic tonsillitis and/or hypertrophy of palatine tonsils in preschool children suffering from lymphoproliferative syndrome is ensured by the palatine tonsils debridement. An interleukin-1β(IL-1β) level is measured in the palatine tonsils washing. If the measured value is less than 5.8 pg/ml, recombinant interleukin-1β (IL-1β) is to be administered orally by phonophoresis with the use of the Tonsillor MM apparatus. Two courses of 10 procedures every 14 days are performed. The clinical effectiveness is assessed if observing a positive dynamics of IL-1β measured in the palatine tonsils washing 17 and 41 days after the beginning of the immunomodulatory therapy.

EFFECT: higher clinical effectiveness ensured by the differentiated selection of children for carrying out the immunomodulatory therapy, reducing a rate of infectious involvements of the palatine tonsils in the declared group of patients by the pathogenetically reasoned application of recombinant IL-1β.

3 cl, 2 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: implementing the method involves determining the content of biomarkers - matrix metalloproteinase 2 (MMP 2) and tissue inhibitor of metalloproteinase 1 (TIMP 1) in the patient's oral fluid sampled before or not earlier than 30 minutes following meals. Using the derived values enables diagnosing papilloma or cancer of patient's oral floor mucosa. Using the invention provides achieving a considerably higher accuracy of the differential diagnosis of benign and malignant new growths of the patient's oral floor mucosa on the day of vising, a considerably higher sensitivity of the differential instant diagnosis, a higher probability of detecting the processes of benign and malignant new growths in the patient's body both at the early stages of the disease, and at the later ones, as well as a considerably simpler preparation of the patient within certain time frames for sampling and storing the diagnostic material.

EFFECT: lower loss of sensitivity of the detection channel in the presence of multiple non-synchronous pulse noise and mutual interference.

3 cl, 5 ex

FIELD: medicine.

SUBSTANCE: polymerase chain reaction method is used to recognise polymorphous variants of IL6 and TGFb1 genes. Recognising the homozygous genotype CC in -174 position of IL6 gene in males and females, as well as the heterozygous genotype GC in -915 position of TGFb1 gene in females enables predicting the high risk of the complicated clinical course of the urogenital Chlamydial infection.

EFFECT: invention enables deciding on reasonable grounds on selecting a therapeutic approach to a specific patient suffering from urogenital Chlamydial infection in order to prevent complications of the urogenital Chlamydial infection and reproductive dysfunctions.

4 tbl, 5 ex

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