Method for prediction of developing atopic diseases in newborns
SUBSTANCE: for the purpose of prediction of developing atopic dermatitis in newborns, umbilical blood plasma is examined for a level of interleukin-18. If the related level is 34 pg/ml or lower, developing atopic dermatitis in the newborn is predicted.
EFFECT: use of the given method enables well-timed evaluation of a risk of developing atopic dermatitis in newborns and early preventive measures for prevention thereof.
The invention relates to medicine, namely to Pediatrics and Allergy, and can be used for early prediction of atopic diseases in children.
A known method for predicting the development of atopic dermatitis in infants by definitions in the cord blood of newborns absolute number of mononuclear cells that synthesize interferon-gamma (INFγand without activation marker CD69 (phenotype cells - INFγ+ CD69-). When the value of this index is 5.3 cells/ml and below, predict the development of atopic dermatitis in infants [EN 2330291 C1, publ. 27.07.2008]. The method allows to determine the risk of development of atopic allergic diseases in infants, but is costly and time-consuming to conduct, as it requires the isolation and cultivation of cells.
Closest to the claimed method is a method of forecasting the development of atopic allergic diseases in the newborn, based on the determination of the concentration of total IgE in serum of umbilical cord or peripheral blood of a newborn using enzyme immunoassay [EN 2108749 C1, publ. 20.04.1998]. The known method is insufficiently informative, as it does not reflect the cellular mechanism propensity to atopy. Moreover, the prognostic securities shall be such laboratory criteria as the level of serum IgE ambiguous.
The task of the invention to provide an informative method for predicting the development of atopic diseases in infants.
The technical result is obtained when implementing the claimed invention:
- expanding Arsenal of diagnostic tools providing the ability to predict the development of atopic diseases in infants;
early prediction of the development of atopic diseases in children;
- information content, reliability and safety of the method.
The essence of the claimed method is that in the plasma of umbilical cord blood of a newborn enzyme-linked immunosorbent assay to determine the level of interleukin-18 (IL-18), and when the level is equal to 34 PG/ml or lower predict the development of atopic disease in the newborn.
Interleukin-18 (IL-18) is a regulatory cytokine, similar to the cytokine family, IL-1 (on the structure, nature of the reception, the type of signal, anti-inflammatory properties), as well as with IL-12 (as inducer of Th1 cytokines profile and as a stimulator of T-cell cytotoxicity). Stimulates production of interferon-gamma by Th1 cells type, activates the EC and monocytes, has a strong influence on the processes of proliferation and cytotoxicity of T-lymphocytes and EC compared with IL-12 [Found on the Internet at: http://allimmunology.org/
immunologicheskij-slovr/i/interlejkin-18-19-20-i-21-il-18 il-19 il-20 il-21]. It is known that IL-18 is a pleiotropic, constitutively expressed cytokine-containing substances that occur in many cell types, mainly by activated monocytes and dendritic cells. The main effects of IL-18 is directed to stimulation of the production of such cytokines as γ, α, IL-1β, as well as adhesion molecules and factors that stimulate apoptosis. Inhibits the activity of cells of the Th2 type. IL-18 is most actively involved in anti-infective and anti-tumor defense of the organism. Shown the protective role of IL-18 in the mechanisms of development of atopic diseases, which led to the choice of this index as a marker of risk for development of atopic diseases in children [Peter J. Barnes. 2008. The cytokine network in asthma and chronic obstructive pulmonary disease. The Journal of Clinical Investigation. 118: 3546-3556. Found on the Internet at: http://www.jci.org/articles/view/36130].
Threshold value (34 PG/ml) was obtained empirically on the basis of a comparative study of research data on the content of interleukin-18 in umbilical cord blood of newborns. Studies have shown that the level of IL-18 is equal to 34 PG/ml (border 25 quartile in the sample authors) or lower risk for development of atopic disease in the newborn should be considered high.
The method is as follows. At birth produce fence 1-2 ml of umbilical cord blood, which is the center of fugeret, then select the plasma in the amount of 0.5 ml of the prepared plasma determine the level of IL-18 enzyme-linked immunosorbent assay using commercially available reagents, for example, companies Bender Medsystems. When the level of IL-18 is equal to 34 PG/ml or lower predict the development of atopic allergic diseases in the newborn. These children belong to the risk group for the development of atopic diseases.
Under the proposed method surveyed 57 infants. 20 children identified low levels of IL-18 (≤34 PG/ml), which gave rise to attribute them to the risk of development of atopic diseases. At the same time the mothers of these newborns in history registered simultaneously atopic dermatitis and clinical manifestations of allergic rhinitis (mother with the presence of atopic phenotype).
Example 1. Newborn female. Studied umbilical cord blood on the proposed method. ELISA determined the level of IL-18 of 22.44 PG/ml (below 34 PG/ml). Newborn baby, according to the invention, is classified as high risk for development of atopic diseases.
Example 2. A newborn male. Studied umbilical cord blood on the proposed method. ELISA determined the level of IL-18 - 47,22 PG/ml (above 34 PG/ml). Newborn baby, according to the invention, is classified as low risk developed by the Yu atopic diseases.
The use of the invention allows to determine the risk of development of atopic allergic diseases in infants and conduct early preventive measures for their prevention.
A method for predicting the development of atopic diseases in infants by study of umbilical cord blood enzyme-linked immunosorbent assay, characterized in that in the plasma of umbilical cord blood of a newborn determine the level of interleukin-18 and when the level is equal to 34 PG/ml or below, predict the development of atopic disease in the newborn.
SUBSTANCE: method under the invention provides that the complex immunoglobulin preparation containing a component of C3b complement is sorbed in microplate wells for the purpose of immunoassay. Then the microplate wells are added with a solution of an analysed sample containing human complement C1 inhibitor with the unknown activity. It is followed by incubation, drying and washing of the dish; the wells are added with a conjugate of enzyme with serine proteinase in the form of preparation of fibrinolysin and a substrate of this enzyme. The amount of the prepared enzymatic reaction product is used to derive the content of active C1 inhibitor. A kit for implementing the method comprises a flat-bottomed microplate with bound C3b, the conjugate of enzyme with serine proteinase, the substrate buffer of this enzyme and a reference for active C1 inhibitor.
EFFECT: use of the method under the invention enables determining activity of C1 inhibitor, simultaneously bound both with serine proteinase inhibition, and with binding inhibition of complement factor B and C3b complement component.
2 cl, 1 tbl, 1 dwg
SUBSTANCE: method under the invention provides that the preparation pyrogenal is sorbed in microplate wells for the purpose of immunoassay; then the microplate wells are added with a solution of an analysed samples containing complement human C1 complement inhibitor with the unknown concentration. Then the plates are incubated, and after washing and drying, a conjugate of enzyme with C1 inhibitor antibodies and a substratum of this enzyme are added into the wells. The amount of the prepared enzymatic reaction product enables calculating the content of the C1 inhibitor in the analysed sample. A kit for implementing the method comprises a flat-bottomed microplate with the sorbed preparation of pyrogenal, the conjugate of the enzyme with the human C1 inhibitor antibodies, a substrate buffer of this enzyme and a reference for the C1 inhibitor.
EFFECT: use of the method enables implementing quantitation of the C1 inhibitor using an ability of the latter to bind to lipopolysaccharide.
2 cl, 1 dwg, 2 ex
SUBSTANCE: method under the invention provides that the preparation heparin is sorbed in microplate wells for the purpose of immunoassay. Then the microplate wells are added with a solution of an analysed samples containing complement C1 inhibitor with the unknown concentration. It is followed by incubation, and after washing and drying of the dish, a conjugate of enzyme with C1 inhibitor antibodies and a substratum of this enzyme are added into the wells. The quantitation is enabled by the amount of the prepared enzymatic reaction product. A kit for implementing the method comprises a flat-bottomed microplate with the sorbed preparation heparin, the conjugate of the enzyme with the human C1 inhibitor antibodies, a substrate buffer of this enzyme and a reference for the C1 inhibitor.
EFFECT: use of an ability of the C1 inhibitor to bind to heparin to determine its content in the analysed samples.
2 cl, 2 dwg, 2 ex
SUBSTANCE: pharmaceutical preparation CIP (complex immunoglobulin preparation, consisting of IgG, IgA and IgM) is bound in cavities of a micropanel, said preparation containing a C3b complement component, samples containing the determined functionally active human C1 inhibitor are incubated in the cavities, and the bound C1 inhibitor is determined using a conjugate of an antibody against the human C1 inhibitor with an enzyme and a substrate for that enzyme based on the amount of the formed product of the enzymatic reaction. The set contains flat-bottomed micropanel with the sorbed CIP, the enzyme conjugate with antibodies against the human C1 inhibitor, a substrate buffer of that enzyme and a standard for the active C1 inhibitor.
EFFECT: use of the method enables to determine activity of the C1 inhibitor which regulates the complement alternative path.
2 cl, 1 dwg, 1 tbl, 2 ex
SUBSTANCE: complex antigen of measles virus used as a component of immunoenzymometric test system for diagnostics of antibodies to measles virus was produced from culture liquid containing measles virus stain Leningrad 16 with titer no less than 105.0 50% tissue cytopathic dose/ml, Edmonston - no less than 106.0 50% tissue cytopathic dose/ml, NovO/96 no less than 108.0 50% tissue cytopathic dose/ml by inactivation of its infectious activity by detergent and separation of protein from cellular lysate by chromatographic purification in amount no less than 2 mcg/ml with the purity no less than 70%. Culture virus-containing liquid was processed using separate cultivation of measles virus stains Leningrad 16, Edmonston and NovO/96 on monolayer of Vero cells culture with subsequent mixing of culture virus containing liquid in proportion 1:1:1 v/v.
EFFECT: Usage of invention provides for increased sensitivity and specificity of a complex antigen possessing the property to detect antibodies in blood serum.
2 cl, 4 tbl, 6 ex
FIELD: chemistry; biochemistry.
SUBSTANCE: invention concerns medical virology and microbiology. Strain is deposited in culture collection of Federal State Research Institution State Research Centre of Virology and Microbiology "Vektor" of Rospotrebnadzor, under registration No VB-05. Strain features higher productivity. More sensitive immunoenzyme test system for hepatitis virus antibody diagnostics is created on the basis of this strain. Invention can be applied in virology.
EFFECT: production of more sensitive immunoenzyme test system for hepatitis virus antibody diagnostics.
2 cl, 1 dwg, 1 tbl, 4 ex
FIELD: medicine; ophthalmology.
SUBSTANCE: invention is intended for simultaneous verification of uveal melanoma and forecast of metastasis development. Immunohistochemical analysis of protein S-100 and melanin-A expression within tumour cell is accompanied with simultaneous estimation of S-100-positive cells number within visual field. Uveal melanoma is verified if reaction of S-100 and melanin-A is positive with forecasted high possibility of tumour dissemination in case number of S-100-positive cells number is less than 50.
EFFECT: simplified examination associated with high specificity and sensibility of simultaneously verified uveal melanoma and forecasted metastasis.
1 tbl, 1 dwg, 2 ex
FIELD: medicine, veterinary.
SUBSTANCE: specific antibodies to virus of goose enteritis are determined by IEA where virus received following injection of epizootic strain "P-75" of enteritis virus, is used as an antigen, and macro porous glass with diameter not less than 700 is used for purification of virus-containing material, the goose Ig G specific immunoperoxidase conjugate is used as an anti-specific agent, and the IEA results are calculated by use of the formula: titer = antiIg[2.02(lg S/P)+3.51], where S is for optical density of tested serum; P is for optical density of the positive control.
EFFECT: method reduces the test time and economical costs.
2 tbl, 2 ex
SUBSTANCE: method involves determining total IgA quantity using immunoenzyme method based on polyclonal IgA antibodies as binding antibodies on the micropanel and the same antibodies in conjugate with peroxidase for detecting bound IgA in sample before and after treatment with specific IgA1-protease. Total IgA1 and IgA2 content is determined before being treated with enzyme. IgA2 content only is determined after being treated with enzyme. IgA1 is not determined after being disintegrated with IgA1-protease. Kit has microplate containing absorbed polyclonal IgA antibodies, conjugate of the same antibodies with peroxidase, reference sample having given IgA concentration, IgA1-protease and substrate buffer.
EFFECT: simplified method and kit for determining IgA1 and IgA2; improved results reproducibility; no specific monoclonal antibodies being used.
2 cl, 1 tbl
SUBSTANCE: method involves absorbing myeloma IgA1 on micro panel, incubating solutions containing IgA1-protease and determining enzyme activity from IgA1 sorbate quantity reduction determined by conjugate with polyclonal IgA antibodies peroxidase. Determining IgA1-protease activity with inhibitors available in various concentrations enables one to calculate its inhibition constants. Kit has microplate containing absorbed myeloma IgA1 preparation, peroxidase conjugate with antibodies against human IgA and substrate buffer.
EFFECT: simplified method and kit for determining IgA1-protease activity and studying its inhibition process.
2 cl, 1 dwg, 1tbl
FIELD: medicine, ophthalmology.
SUBSTANCE: in lacrimal liquid one should detect the content of interleukin 8 (IL-8) and that of interleukin 1 beta (IL-1β) to calculate prognostic coefficient (PC) due to dividing the first value by the second one by the following formula: At PC value being below 10.0 one should predict favorable disease flow, and at PC value being above 10.0 - unfavorable flow.
EFFECT: higher accuracy of prediction.
FIELD: medicine, ophthalmology.
SUBSTANCE: one should detect the content of tumor necrosis factor alpha in acute period, moreover, the above-mentioned factor should be determined in lacrimal liquid on the 11th - 15th d against the onset of herpetic ophthalmoinspection, at its content ranged 176-250 pcg/ml prediction is considered to be favorable, and from the value of 300 pcg/ml and higher - as unfavorable.
EFFECT: higher accuracy of prediction.
FIELD: medicine, surgery.
SUBSTANCE: one should carry out virological testing patient's blood serum and hepatic bioptates. At detecting TTVDNA and HGVRNA it is necessary to perform ultrasound survey, and at availability of biliary sludge one should conclude upon early stage of cholelithiasis.
EFFECT: higher accuracy of diagnostics.
FIELD: medicine, cardiology.
SUBSTANCE: one should detect the level of activity of IgM and IgG immunoglobulins to cytomegalovirus on the 5th and 15th d of large-focus myocardial infarction. At increased diagnostically valuable result of specific immunoglobulins of type M by 0.10-0.20 times and for type G by 0.73-2.09 times it is possible to predict favorable clinical flow of large-focus myocardial infarction without exudative pericarditis. At the value of specific immunoglobulins exceeding diagnostically valuable result for type M - by 0.86-1.67 times and for type G by 2.42-3.01 times one should predict early clinical flow of post-infarction pericarditis. The method enables to carry out prophylactic measures in due time.
EFFECT: higher accuracy of prediction.
5 ex, 2 tbl
SUBSTANCE: one should test blood serumal samples in immunoenzymatic assay by applying a test system ELI-N-1, the components of which are being antigens of nervous tissue or their immunochemical analogs specifically binding antibodies with tendency to antigens of nervous tissue, and, also, idiotypical antibodies to them, or their variable parts. Prediction should be performed according to the level of idiotypical and anti-idiotypical autoantibodies to antigens of nervous tissue and, also, according to their ratio. Thus, a new test system has been elaborated that enables to predict the flow of available nervous-psychic diseases.
EFFECT: higher accuracy of prediction.
2 cl, 7 ex
FIELD: medicine, obstetrics.
SUBSTANCE: in pregnant women at pregnancy terms being after 39 wk in average portion of morning urine one should detect due to a solid-phase immunoenzymatic assay the concentration of pregnancy-associated protein-A (PAPP-A). At the level of PAPP-A being 768.2 ng/ml and more it is possible to conclude upon physiological mature pregnancy. The innovation is noninvasive and enables to increase accuracy in predicting true over-mature pregnancy.
EFFECT: higher efficiency and accuracy of diagnostics.
2 ex, 1 tbl
FIELD: medicine, biotechnology.
SUBSTANCE: the present innovation deals with elaborating diagnostic reagents for testing prionic protein in mammalian cerebral tissue due to IEA technique and refers to antibodies specifically reacting with prionic protein PrP or its fragment. The innovation includes polyclonal antibodies obtained to synthetic peptides including amino acid sequences of bovine prionic protein 143-168, 101-134 and 211-241, their conjugates with horseradish peroxidase and diagnostic reagents obtained upon their basis that enable to detect prionic protein in mammalian cerebral tissues.
EFFECT: higher specificity of detection.
15 cl, 8 ex, 8 tbl
FIELD: veterinary and medicine.
SUBSTANCE: invention, in particular, relates to production and use of biological preparations intended for differential diagnostics of brucellosis and to a method of differentially diagnosing brucellosis. Method involves serologic analysis of sera using antigenic "IFK", which is horse-radish peroxidase-labeled electrophoretically purified polypeptide fraction of virulent strain B.arborus 54. Diagnosis of brucellosis is stated when anti-brucellosis antibodies in sick animal sera diluted to 1/100 and higher are revealed at CSP reaction intensity 2.1 and higher.
EFFECT: elaborated method increasing immuno-enzymatic test specificity and allowing performing differentiation of postvaccinal immunological reactions and postinfectious reactions induced by microorganisms having antigenic affinity with brucellas, especially Yersinia enteriocolitica.
2 cl, 4 ex
FIELD: biotechnology, analytical chemistry.
SUBSTANCE: claimed method includes providing of complexes between antigen molecules and specific antibodies on carrier surface, wherein said complexes are disclosed by addition of enzyme label thereto followed detection thereof based on formation of enzyme reaction product. Enzyme label addition is carried out by two protein interaction, namely bacterial ribonucleaze barnase and barstare, which is inhibitor thereof, wherein either of the two is added to immunoreagent, and the other one is added to enzyme label. Abovementioned complexes have high affinity, specificity, and binding constant of 1014 M-1.
EFFECT: new method for antigen detection.
6 dwg, 2 ex
FIELD: veterinary virology.
SUBSTANCE: the present innovation deals with interaction of antibodies with an antigen, with antibodies labeled with horseradish peroxidase, addition of substrate mixture and registration of reaction results. Moreover, one should apply plotting boards with presorbed antibodies and general immunoenzymatic conjugate. The innovation enables to shorten terms for diagnosis and obtain more significant results of diagnostics.
EFFECT: higher efficiency.