Method for pre-operative prediction of complicated course of early post-transplantation period manifested by transplant dysfunction following living-related liver transplantation in infants with congenital hepatobiliary diseases
SUBSTANCE: preoperative prediction of the complicated course of the early post-transplantation period is ensured by measuring the pre-transplantation blood plasma concentration of insulin-like growth factor-1 (IGF-1) and growth hormone (GH), ng/ml. That is followed by calculating a prediction index for the post-transplantation course (K) by formula: K=lg (IGF-1/GH). If K is less than minus 1.7, the complicated course of the early post-transplantation period is predicted.
EFFECT: using the method enables the objective prediction in the infants suffering congenital hepatobiliary diseases accompanying the early post-transplantation period manifested by transplant dysfunction.
The invention relates to medicine, namely to organ transplantation and clinical laboratory diagnostics, can be used for pre-operative predicting the course of the early post-transplant period in young children after liver transplantation for congenital diseases of the hepatobiliary system.
The problem of predicting the course of the early postoperative period in children of early age after liver transplantation is essential due to the fact that this category of patients refers to patients with a high risk of complications and adverse outcome, as evidenced by the numerous publications of foreign and domestic authors.
The peculiarity of infants with congenital hepatobiliary disease, which is related liver transplantation is a combination of factors, multidirectional influence on the prognosis of treatment effectiveness, for example: due to the age of Hyper-reactivity of the immune system and at the same time, high ability to develop immunological tolerance to the graft. The combined effect of these factors is difficult to assess. Dysfunction of the graft in the early postoperative period may be due to several reasons: immunological, infectious, metabolic, SOS is done and other It is believed that young children after liver transplantation prevails the risk of infectious complications, against which there is a necessity of correction of the standard Protocol immunosuppressive therapy (deferred the appointment of immunosuppressants, their temporary cancellation, lower doses of immunosuppressants).
Safety minimization of immunosuppressive therapy in the early postoperative period, as the need for more careful preoperative preparation of patients (strengthening antibacterial, antiviral therapy, correction of metabolic status) can be estimated taking into account the risk of complications during the early postoperative period. That is why there is a need in the way of forecasting the preoperative stage of the complicated course of the early post-transplant period due to graft dysfunction in children of early age related liver transplantation.
A method for the prediction of 6-month survival of patient and graft after liver transplantation for children with congenital diseases of the hepatobiliary system (McDiarmid SV, Anand R, Martz K, et al. A multivariate analysis of pre-, peri-, and post-transplant factors affecting outcome after pediatric liver transplantation. Pediatric Transplant. 2011 Jun; 15 (4): 400-5) the Essence of the method lies in the fact that, with the development in the early post-transplantation per the ode complications such as vascular thrombosis, intestinal perforation, septicemia, predict a high risk of death or graft loss within the next 6 months. However, this method allows to predict the survival of recipient and graft on the basis of the analysis of postoperative events.
In the prior art there is also known a method of pre-operative prediction of the development of graft dysfunction in infants with congenital diseases of the hepatobiliary system (Shevchenko O. P., Hickon O. E., Kuntsevich N. In., and other Communication between sCD40L level and nature of the current post-transplant period in the recipient liver. Bulletin of Transplantology and artificial organs. App. The materials of the V all-Russian Congress of transplantologists / Moscow, 2010, S. 126-127), which is chosen as a prototype.
It consists in the following. In the blood plasma of children of candidates for liver transplantation, using the ELISA method to measure the concentration of a soluble form of CD40 ligand (sCD40L) and identifying the source of the high level of sCD40L (>3.3 ng/ml) predict the development of graft dysfunction.
However, this method allows to predict a narrow range of complications, such as graft dysfunction, manifested by immunological mechanisms, because of the co-stimulatory signal system CD40-CD40L plays important is Yu role in the mechanisms of immune regulation, in particular, activation of T-lymphocytes, antigen presenting cells and endothelial cells.
The present invention is a pre-operative prediction of complications during the early post-transplant period due to graft dysfunction in children of early age after liver transplantation for congenital diseases of the hepatobiliary system.
The technical result of the invention is the possibility of accurate preoperative prediction of complications during the early postoperative period, manifested by graft dysfunction of various etiologies in children of early age after liver transplantation for congenital diseases of the hepatobiliary system, which would take into account the integral indicator of metabolic and immune mechanisms in the development of dysfunction of the graft liver, while maintaining the simplicity, availability of study do not require a long time, unique reagents and instruments.
The essence of the proposed method for the preoperative prediction of complications during the early postoperative period, manifested by graft dysfunction after liver transplantation in young children with congenital diseases of the hepatobiliary system is as follows.
Before transplantation, pécs and in the blood plasma of candidates for transplantation to determine the concentration in ng/ml insulin-like growth factor-1 (IGF-1) and somatotropic hormone (STH). Next, calculate the index of forecasting according To the formula: K=lg (IGF-1/STH). When the value of K less minus 1.7, predict complications during the early post-transplant period, manifested by dysfunction of the graft.
We have empirically found that a certain ratio of the concentrations of IGF-1 and STG in plasma before the operation, expressed by the decimal logarithm, is an informative predictor of the course of the early post-transplant period after related liver transplantation in young children with congenital diseases of the hepatobiliary system.
Congenital liver diseases are accompanied by deficiency of IGF-1, increasing as the progression of the disease and reduce belkovosinteticescuu liver function. In response to the deficiency of IGF-1 on the mechanism of negative feedback there is an increase in the secretion of STH, one of whose functions is to stimulate the synthesis of IGF-1. Thus, as the progression of diseases of the hepatobiliary system increases the deficit IGF-1 and increases the concentration of STH. Both hormones have an effect on the metabolic status of the patient and the state of his immune system. It turned out that a pronounced imbalance between IGF-1 and STG before liver transplantation has an adverse influence on the course early in reoperations period and increases the risk of graft dysfunction, that may be associated with metabolic and immune mechanisms and is a reliable prognostic indicator. Thus, we found the index forecasting, assessing the imbalance in the system STH - IGF-1, allows us to predict a wider spectrum of complications in the early post-transplantation period.
The applicant is not aware of the application of the logarithm of IGF-1/STH with the aim of predicting the course of the early postoperative period after related liver transplantation.
The method is as follows.
Measurement of IGF-1 and STG conduct quantitative immunoassay method ELISA reagents IDS IGF-I ELISA, USA, and Diagnostic Biochem Canada Inc., respectively (see, for example, instructions to the kit reagents IDS IGF-I ELISA and assay kit Growth hormone, Diagnostic Biochem Canada Inc; B. C. Kamyshnikov. Handbook of clinical-biochemical laboratory and laboratory diagnosis / Moscow, Medpress-inform, 2009, 371 S.; Liss C. L., Nikolaev, L. C., Nagorno I. I. et al. Laboratory diagnostics and functional test in pediatric endocrinology. SPb.: Special literature, 1996. - 136 C.) on the photometer Zenyth 340 (Anthos Labtec Instruments GmbH, Austria).
If the index patient To less than minus 1.7 to operations, predict complications during the early postoperative period, manifested on what funkcia transplant.
The possibility of practical use of the method of achieving the specified purpose and the technical result is proved the following data.
Example 1. The patient W-VA, 7 months, (b 1892/2011) was admitted with a diagnosis of cirrhosis of the liver in the clinical outcome of bailer with syndromes of liver cell failure, portal hypertension (splenomegaly, ascites), cholestasis. When entering the weight 6.5 kg (5-15 percentile), height 67 cm (25-50 percentile). Score PELD 24,6. Before the surgery, the concentration of IGF-1 in plasma of 0.01 ng/ml, STH and 4.4 ng/ml, the index K=-2,7. The forecast adverse trends early postoperative period. The patient underwent an operation orthotopic transplantation of the left lateral sector of the liver from a living related donor, cousin, grandmother, 44 years. The postoperative period for 15 days was uneventful. Immunosuppressive therapy was assigned according to the standard scheme. On the 16th day post-operative period was complicated by formation of a partial external biliary fistula. In further noted the development of bacteremia, the formation of septic conditions that demanded the cancellation of immunosuppressive therapy. Laboratory indicated an increase in the activity of alanine aminotransferase and aspartate aminotransferase, increased concentrations of total and direct bilirubin and decrease the end of the ation of albumin and total protein in blood plasma. On the background of multiple changes, antibacterial Protocol normalized body temperature, decreased leukocytosis in the blood, blood cultures became sterile, however, continued to grow manifestations of dysfunction of the graft. Given the long-term abolition of immunosuppressive therapy on the background of the septic condition, the clinical picture was interpreted as a rejection of the graft liver. Conducted in vitro detoxification (low flow veno-venous ultradilute). Reported a temporary positive dynamics of biochemical parameters of blood. However, the patient's condition continued to deteriorate, there was a growing signs of respiratory, cardiovascular, hepatic, renal insufficiency. The patient died on the 35th day after the operation. According to the results of the postoperative period prediction was confirmed.
Example 2. The patient On s, 12 months (and/b 1659/2010), was admitted with a diagnosis of cirrhosis in the outcome of congenital malformation of the biliary tract with symptoms of cholestasis, hepatic cell failure, portal hypertension (splenomegaly, ascites, varices esophageal 1-2 degrees), protein-energy malnutrition (underweight 18%); after portoenterostomy by Kasai, recurrent cholangitis. The PELD score of 21.7. Before the surgery, the concentration of IGF-1 in plasma is 0.001 ngml, STH - 12,25 ng/ml, the index K=-4,1. Was made unfavorable prognosis of early post-operative period. The patient performed orthotopic transplantation of the left lateral sector of the liver from a living related donor, father, 24 years. Immunosuppressive therapy is assigned according to the standard scheme. During the early postoperative period was complicated by formation of a partial external biliary fistula. In connection with the occurrence of fever was canceled tacrolimus, and then, after 7 days, became a marked increase of transaminase activity. Immunosuppressive therapy was resumed in full, which led to normalization of biochemical parameters of liver function. Incomplete outer fistula independently closed at 53-and the day after surgery. The patient was discharged with satisfactory graft function on the 56th day after the operation. The prediction was confirmed in relation to the current early postoperative period.
Example 3. The patient In society, 27 months (and/b 1479/2011), was admitted with a diagnosis of cirrhosis in the outcome of biliary atresia with syndromes of liver cell failure, cholestasis. Portal hypertension (splenomegaly, varices esophageal 3-4 St); after portoenterostomy by Kasai; physical development below average, harmonic. Score PELD 7(forecast 3-month survival without transplantation favorable). However, before the surgery, the concentration of IGF-1 in plasma - 0,0068 ng/ml, STH - 31.5 ng/ml, the index K=was 3.7. The prediction was made adverse trends early post-transplant period. The patient performed orthotopic transplantation of the left lateral sector of the liver from a living related donor, grandmother, 53 years. During the early postoperative period was complicated by formation of a partial external biliary fistula, right upper lobe pneumonia, CMV infection, thrombosis of the portal vein. Against the background of these complications were temporary elimination of immunosuppressive therapy, which was accompanied by increased activity of transaminases, increased coagulopathy. Was renewed monotherapy immunosuppressants tacrolimus (therapy with methylprednisolone was not renewed due to long-lasting outer incomplete biliary fistula). On the background skorrigirovanna therapy showed improvement of the child's condition. The patient was discharged on the 70th day after the operation with satisfactory performance of the functions of the liver transplant. For during the early postoperative period, the prediction was confirmed.
Example 4. Patient G s, 7 months (and/b 1821/2010), was admitted with a diagnosis of biliary cirrhosis in the outcome of atresia extrahepatic bile ducts, hepatic syndromes-cell failure, then what happens hypertension (varicose veins of the esophagus 2-3 article, splenomegaly, ascites); after portoenterostomy on Kasai. Score PELD 21. Before the surgery, the concentration of IGF-1 in plasma or 0.035, STH with 4.65 ng/ml, the index K=of-2.1. The prediction was made adverse trends early post-transplant period. The patient performed orthotopic transplantation of the left lateral sector of the liver from a living related donor, mother, 34 years. Immunosuppressive therapy was carried out according to the standard scheme, tacrolimus assigned on day 7 after surgery. To the 15 th day after the operation - smooth postoperative period. With a 15-day was marked increase of transaminase activity, according to the survey, there were no vascular, biliary, or infectious complications. Was held pulse therapy with corticosteroids, with a positive effect, therefore, a conclusion was made about the immunological character of transient dysfunction of the graft. For during the early postoperative period, the prediction was confirmed.
Example 5. The patient-s, 7 months (and/b 613/2010), was admitted with a diagnosis of cirrhosis in the outcome of biliary atresia, with the syndrome of portal hypertension (ascites, hepatosplenomegaly), hepatic cell failure; status post cholecystectomy, portoenterostomy on Kasai. The patient noted a pronounced retardation times the development (weight 6,7 kg less than 3 percentile, growth of 55 cm, less than 3 percentile). Score PELD at entry 36. On the eve before surgery in plasma were measured concentration of IGF-1 is 13.4 ng/ml and STG - 1,67 ng/ml, index To 0.9. The patient was taken to the favorable prognosis of early postoperative period, despite the severity of liver failure. The patient was performed orthotopic transplantation of the left lateral sector of the liver from a living related donor, mother, 26 years. During the early postoperative period was without complications. The drains are removed within the first three weeks after surgery. Immunosuppressive therapy was carried out on a standard two-part Protocol. The child was discharged on the 23rd day after the operation. For during the early postoperative period, the prediction was confirmed.
Example 6. Patient T-VA, 4 months (and/b 376/2010), was admitted with a diagnosis of cirrhosis in the outcome of biliary atresia with syndromes of liver cell failure, portal hypertension (hepatosplenomegaly, ascites, varices esophageal 3-4 degrees). The PELD score of 15.7. Before the operation, the concentration of IGF-1 in plasma - 79 ng/ml, STH - to 33.9 ng/ml, the index K=0,37. Was made a favorable prognosis early postoperative course. The patient was performed orthotopic transplantation of the left lateraling the sector liver from a living related donor, mother, 37 years. During the early postoperative period smooth. The patient was discharged on the 32nd day after surgery with satisfactory function of the graft liver.
The proposed method is tested in clinical practice in 39 patients, and the prognosis of early post-transplant period, made by the claimed method was confirmed in 33 cases, which amounted to 84.6%.
Use in clinical practice patentable method allows a rational approach to the selection and more thorough preparation of patients for surgery, as well as the establishment of appropriate therapy protocols for selected risk groups, which suggests the possibility of increasing the effectiveness of treatment.
The way the preoperative prediction of the complicated course of the early post-transplant period, manifested by dysfunction of the graft, after a related liver transplantation in young children with congenital diseases of the hepatobiliary system, including the study of blood plasma of the recipient before the operation, characterized in that in the blood plasma was measured concentration in ng/ml insulin-like growth factor-1 (IGF-1) and somatotropic hormone (STH), and complications during the early post-transplant period, manifested by dysfunction of the graft, predict when meant and lg (IGF-1/STH) less minus 1.7.
FIELD: veterinary medicine.
SUBSTANCE: method of diagnostics of leptospirosis in farm animals, comprising determining the presence of antibodies to antigens of seven serogroups L.hebdomadis, L.pomona, L.tarassovi, L.grippotyphosa, L.canicola, L.sejroe and L.icterohaemorrhagiae in blood serum by the method of enzyme immunoassay, characterised in that the antigen is used as the antigenic proteins of Leptospira of three serogroups mixed in equal quantities - L.tarassovi, L.grippotyphosa and L.hebdomadis. The invention significantly simplifies and minimizes the diagnostics of leptospirosis, enables to implement the operational control of epizootic condition of the animals on leptospirosis.
EFFECT: method has a high specificity, sensitivity and commonality, provides creation of a safe labour conditions for the staff of diagnostic laboratories.
4 tbl, 4 ex
SUBSTANCE: technique according to the invention involves the two-stage immobilisation of S and R brucellosis antigens on a tray by a non-specifically adsorbed mouse's monoclonal antibodies 2H2 and 2H8 in wells of the immunological tray (the first stage), specific binding S and R brucellosis antigens (the second stage) thereto, introduction and incubation of the analysed material (blood serum or milk) combined with introduction of mixed mouse's monoclonal antibodies 2H2 and 2H8 marked by horseradish peroxidase. The brucellosis agent antibodies are detected by being competitive with the antigen in binding on the well surfaces of the tray between the antibodies of the analysed sampled and fragment-marked monoclonal antibodies. A decreased level of the enzymatic signal in the analysed sample as compared with the negative reference testifies to contamination of the animal. Introducing the analysed sample into the two wells of the tray with S and R brucellosis antigens enables differentiation thereof for antibody specificity to any given form of the disease agent.
EFFECT: method involving the enzyme immunoassay enables increasing the effectiveness of health-improving measures, reducing the length of health-improvement in livestock farms with the negative brucellosis situation, dropping the disease incidence.
3 tbl, 3 ex
SUBSTANCE: in order to predict severity of postoperative period course in patients with calculous cholecystitis level of hormone grelin is determined in patients' blood serum before operation. If value of grelin level is 0.8-1.5 ng/ml mild course of postoperative period is predicted, if value of grelin level is >1.5-10.0 ng/ml course of medium severity is predicted. And if grelin level in blood serum increases to >10.0 ng/ml, severe course of postoperative period is predicted.
EFFECT: method makes it possible to increase accuracy of prediction of postoperative period course severity in patients with calculous cholecyctitis in cholecystectomy in order to perform treatment correction.
FIELD: veterinary medicine.
SUBSTANCE: method includes the interaction of antigens with antibodies, with anti-species antibodies labeled with horseradish peroxidase, adding the substrate mixture and recording the results of the reaction on the colour intensity of the complex formed. In the reaction the plates are used with antigens of rotavirus, coronavirus, the virus of diarrhea preliminary sorbed on them, and after applying the test samples of serum 0.10-0.12 ml each. The plates were incubated for 1.5-2 h at 36-37°C and washed. Then total anti-species immunosorbent conjugate is applied of 0.10-0.12 ml consisting of enzyme-labeled antibodies against globulins of blood serum of cattle and the results of the reaction are recorded.
EFFECT: invention provides simultaneous diagnosis of rotavirus, coronavirus enteritis, viral diarrhea in cattle, sensitivity, specificity, and ease of the assay, and the ability to automate the process of research.
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.
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: 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.