STUDYING THE POSSIBILITY OF USING IMMUNO-ENZY TEST SYSTEM FOR EVALUATING ANTI-PERTUSSIS IMMUNITY

2020 ◽  
Vol 65 (2) ◽  
pp. 90-94
Author(s):  
O. Yu. Sosnina ◽  
A. M. Nikolaeva ◽  
T. V. Vyaznikova ◽  
I. V. Feldblum ◽  
K. A. Subbotina

A research objective - to study the possibility of using the ELISA Anti-K enzyme immunoassay system to evaluate anti-pertussis immunity. A comparative assessment of the content of co-crank antibodies in the blood serum of adults, pregnant women and children 6 years old in the agglutination test, in the test system “Anti-K ELISA” and test systems of foreign production was carried out. The “Anti-K” IFA test system makes it possible to detect the level of specific antibodies to both the whole cell and cell-free pertussis component of the vaccine at any stage of the vaccination cycle. This diagnostic test can be used to determine the tactics of immunization, and to assess population immunity.

2021 ◽  
Vol 74 (10) ◽  
pp. 2585-2587
Author(s):  
Vitaliy V. Maliar

The aim: To study the features of the course of gestation and perinatal outcomes of delivery in women with vitamin D lack. Materials and methods: The article presents the results of studies of the characteristics of the course of pregnancy and delivery outcomes in 50 patients with vitamin D lack compared with a group of 50 somatically healthy pregnant women with normal level of 25 (OH) D. In order to establish a lack of vitamin D in pregnant women in the 10-12, 20-22, 30-32 weeks of gestation electrochemiluminiscence method by using a test system EURIMMUN (Germany) in the blood serum level of 25-hydroxycalciferol (25 (OH) D) in pregnant women. Results: When analyzing the structure of complications in women with vitamin D lack during pregnancy and childbirth we found out that risk of premature birth and premature births dominated among all the complications, respectively (58.0% and 36.0%) against (12.0% and 16.0%), p <0.05. Vitamin D lack in pregnant women is often associated with a wide range of obstetric and perinatal complications, namely: preeclampsia, gestational diabetes, bacterial vaginosis , premature rupture of membranes, placental abruption, abnormal labor activity, fetal distress that required delivery by Caesarean section. Conclusions: An analysis of the course of pregnancy and childbirth in women of thematic groups proved the expediency of an individual approach to the therapy of obstetric pathology among women with vitamin D lack. Despite the level of 25 (OH) D in the blood serum of a pregnant woman of 30 ng / ml and below, it is advisable to prescribe vitamin D for prophylaxes and treatment of Vitamin D deficiency in mother and fetus.


2018 ◽  
Vol 20 (5) ◽  
pp. 647-656 ◽  
Author(s):  
A. N. Glushkov ◽  
K. S. Krasilnikova ◽  
E. G. Polenok ◽  
M. V. Kostyanko ◽  
R. V. Olennikova ◽  
...  

Specific antibodies against environmental chemical gene toxicants and endogenous steroid hormones are shown to modulate concentrations of these compounds in blood serum and their biological effects in experimental models. However, probable hazards of such antibodies in human teratogenesis are still unknown. In particular, potential correlations between specific serum antibodies, sex hormone levels in pregnant women, and congenital malformations in newborns are not clear. The aim of this study was to identify possible associations between occurrence of antibodies to benzo[a]pyrene, estradiol and progesterone (Bp, Es and Pg, respectively), and congenital malformations, and effects of these antibodies upon Es and Pg concentrations in blood serum of pregnant women. We have included into the study 182 women with normal pregnancy and 101 females with congenital malformations of fetus. A non-competitive solid phase immunoassay was performed using Bp, Es and Pg conjugated to bovine serum albumin as antigens. Es and Pg serum concentrations were measured using immunoassay test-systems of “Immunotech” (Moscow). Results: strong positive correlations were revealed between the levels of studied antibodies in the both groups. High IgA-Bp/IgA-Es (> 3) and IgA-Bp/IgA-Pg ratios (> 3) were associated with congenital malformations (OR = 2.2, p = 0.013 and OR = 6.8, p < 0.0001). Positive correlations were revealed between Pg/Es and IgA-Bp/IgA-Es (rS = 0.62, p < 0.0001), and IgA-Bp/IgA-Pg ratios (rS = 0.77, p < 0.0001) in cases with inborn malformations. Similar correlations were found for the women who had normal pregnancy (rS = 0.4, p = 0.0001, and rS = 0.23, p = 0.026, respectively). The Pg/Es proportion correlated with IgG-Bp/IgG-Es (rS = 0.46, p = 0.002), and with IgG-Bp/IgG-Pg ratio (rS = 0.5, p = 0.0009) in cases of malformations, but not in women with normal pregnancy. Conclusion: we have revealed novel associations between congenital malformations of fetus and ratios of IgA-Bp/IgA-Es, as well as IgA-Bp/IgA-Pg, like as positive correlations between hormonal Pg/Es proportions, and ratios of specific antibodies in pregnant women.


Author(s):  
E. Krieger ◽  
O. Samodova ◽  
L. Titova

Despite high levels of vaccination coverage, pertussis remains a serious problem. Pertussis cases are registered among infants, adolescents, and adults. Infants younger than three months of age have the highest rate of serious clinical course of pertussis. Transplacental transfer of pertussis-specific antibodies induce protection against infection. The available data on antibody level against pertussis among pregnant women in Russia are fragmentary. To evaluate the humoral immunity to Bordetella pertussis in pregnant women and factors associated with seronegative status we performed cross-sectional study including 388 participants. “SeroPertussis IgG” (Israel) enzyme linked immunosorbent assay test-system was used for quantitative measurement of antibodies to pertussis toxin / hemagglutinin. Binary logistic regression analysis was performed to assess factors associated with seronegative status. The median age of the participants was 30 years. The majority of them (51,3%) did not provide documented vaccination against pertussis. A positive (protective) level of anti-pertussis antibodies (10-50 BU/ml) was revealed in the venous blood of 46,9% of pregnant women. In 25,8% of seropositive women, concentration of specific antibodies was high (> 50 BU/ml). That may indirectly indicate recent Bordetella pertussis infection. More than half of the women surveyed (53,1%) did not have a protective titer of antibodies and were considered susceptible to pertussis. Their children will not receive transplacental immunity to infection. Gestational age was significantly associated with seronegative status. Compared to women tested during the first trimester, participants in their third trimester of pregnancy were more likely to be seronegative to pertussis. The odds of being susceptible increased with increased gestational period (p for linear trend < 0,01). There were no significant differences between the seropositive and seronegative participants with regard to age, gravidity and vaccination status. Pertussis booster vaccinations for preschool children, adolescence and healthcare workers dealing with pregnant women and newborns, as well as cocoon vaccination strategy and vaccination during pregnancy, are required to be implemented to protect infants against pertussis.


2022 ◽  
Vol 20 (6) ◽  
pp. 72-80
Author(s):  
L. N. Lukhverchyk ◽  
G. L. Alatortseva ◽  
L. N. Nesterenko ◽  
V. Y. Kabargina ◽  
V. V. Dotsenko ◽  
...  

Relevance. The introduction of Varicella vaccine prophylaxis explains the need to develop a methodology for monitoring the vaccination effectiveness and the intensity of population immunity. This problem can be solved using quantitative immunoassay methods. Aim. Development of an enzyme-linked immunosorbent assay for the concentration of class G immunoglobulins (AB) to Varicella zoster virus (VZV) determining and assessing its functional characteristics and diagnostic efficiency. Materials and methods. Recombinant antigen GE VZV. WHO International Standard for Antibodies to VZV W1044. Blood serum samples from healthy people and patients with Chickenpox and Herpes zoster, blood serum samples containing IgG antibodies to herpes simplex viruses of the first and second types, cytomegalovirus, Epstein-Barr virus. Anti-VZV ELISA (IgG) reagent kit (Euroimmun, Germany). Indirect enzyme-linked immunosorbent assay. Immunization of animals with recombinant antigen GE, isolation, and purification of specific antibodies. Conjugation of monoclonal antibodies to human IgG with antibodies to antigen GE and with horseradish peroxidase. Results. An enzyme-linked immunosorbent assay in «an indirect» format has been developed to determine the specific antibodies to VZV concentration (IU/ml) in human serum/plasma. An artificial calibrator for determining the concentration of AB-VZV had been synthesized and standardized according to the International WHO-standard W1044. The main functional characteristics of the developed enzyme-linked immunosorbent assay are determined in accordance with GOST 51352-2013. The diagnostic kit was tested on blood serum samples from children with chickenpox (n = 43), adults with Herpes zoster (n = 158), healthy individuals (n = 781). The diagnostic sensitivity of the test system was 85%, the diagnostic specificity was 87% according to the ROC analysis. The absence of cross-reactivity of the test system was shown on samples with serological markers of other herpesvirus infections (n = 94). Comparative trials of the developed test system and its commercial analog, the Anti-VZV ELISA (IgG) reagent kit, did not reveal statistically significant differences between their functional characteristics. Conclusions. The developed test system for determining of the AB-VZV concentration in human serum/plasma in terms of its functional characteristics meets the GOST requirements, is characterized by high diagnostic efficiency, can be used to monitor the effectiveness of vaccine prophylaxis and strength of population immunity, as well as to assess the immune response in chickenpox and Herpes zoster.


Author(s):  
V. V Pisarev ◽  
Maria M Ulyashova ◽  
Gelia N Gildeeva

An integral part of preclinical pharmacokinetic studies is the development of a bioanalytical method for determination of the drug in a biological fluid.The aim of the research was to assess the suitability of the test system based on enzyme-linked immunosorbent assay (ELISA) for quantitative determination of rituximab in the blood serum of laboratory animals after intravenous administration of rituximab at a dose corresponding to the therapeutic dose in humans. Th test system was developed by the Scientific and Production Center Probiotech.Materials and methods: the determination of rituximab in biological samples was carried out using a two-stage sandwich-type ELISA, followed by detection based on horseradish peroxidase. The ELISA results were recorded using a microplate photometer at a wavelength of 450 nm.Results: the experiments helped to establish the detection limit (0.24 ng/mL) and the lower limit of quantitation (1.00 ng/mL) of rituximab in rabbit blood serum, they also demonstrated high selectivity of analyte determination in a multicomponent biological matrix. The mean rituximab concentration was within 14 % of the nominal value in the entire working range of the method. The within-run and between-run precision of the assay did not exceed 7.4 %, the total error of the method did not exceed 20.1 %. The linearity of dilution makes it possible to use the assay for the analysis of biological samples with a wide range of rituximab concentrations. The stability of the analyte in the rabbit blood serum was confirmed by storing samples for 6 hours at room temperature, for 50 days at —35 °C, and after 3 freeze-thaw cycles. The validated immunoassay was successfully used to determine the rituximab concentration in biological samples obtained in the rituximab pharmacokinetic trial in rabbits. The accuracy of the results was confirmed for the entire range of the determined concentrations; parallelism was demonstrated between the calibration curve and the results of analysis of serially diluted rabbit serum samples with the maximum concentration of rituximab.Conclusions: the proposed enzyme immunoassay test system can be used for quantitative determination of rituximab in the blood serum of laboratory animals, as it meets acceptance criteria for all validation parameters described in the international guidelines on validation of bioanalytical methods.


2019 ◽  
Author(s):  
Vilius Floreskul ◽  
Fatima Juma ◽  
Anjali Daniel ◽  
Imran Zamir ◽  
Zulf Mughal ◽  
...  

1980 ◽  
Vol 45 (4) ◽  
pp. 1099-1108 ◽  
Author(s):  
Mikuláš Chavko ◽  
Michal Bartík ◽  
Evžen Kasafírek

A polarographic study of the hydrolysis of [8-lysine]vasopressin and some hormonogens of the vasopressin series with the blood serum of women in the last week of pregnancy was studied. The dependence of hydrolysis on pH (pH optimum: 7.4-7.50, substrate concentration (Km 1.2 . 10-5M), pH stability and thermal stability were determined. The rate of hydrolysis of individual vasopressin analogues decreases in the order: [8-lysine]vasopressin > Nα-glycyl-prolyl[8-lysine]-vasopressin > Nα-leucyl-[8-lysine]vasopressin > Nα-alanyl-[8-lysine]vasopressin > Nα-phenyl alanyl-[8-lysine]vasopressin > Nα-diglycyl-[8-lysine]vasopressin > Nα-prolyl-[8-lysine]vasopressin > Nα-triglycyl-[8-lysine]vasopressin > Nα-sarcosyl-glycyl-[8-lysine]vasopressin. The degree of hydrolysis gradually increases to a multiple with the length of the pregnancy in consequence of the presence of oxytocine. However, vasopressin is also hydrolysed to a small extent with the enzymes from the blood sera of non-pregnant women. Under similar analytical conditions oxytocin was not hydrolysed with the sera of non-pregnant women and therefore oxytocin is a more suitable substrate than vasopressin for polarographic determination of serum oxytocinase.


1918 ◽  
Vol 28 (4) ◽  
pp. 449-474 ◽  
Author(s):  
Frederick L. Gates

1. A meningococcus vaccine suspended in salt solution has been given subcutaneously as a prophylactic to about 3,700 volunteers in three injections of 2,000 million, 4,000 million, and 4,000 or 8,000 million cocci at weekly intervals. 2. These doses rarely caused more than the mildest local and general reactions. Exceptionally a more severe reaction emphasized the presence of an unusual individual susceptibility to the vaccine. In such instances the symptoms were in part those of meningeal irritation and sometimes simulated the onset of meningitis. 3. Specific meningococcus agglutinins have been demonstrated in the blood serum of vaccinated men as compared with normal controls. 4. Moreover, agglutinins have been demonstrated in the blood serum of chronic carriers of the meningococcus. Evidence is thus brought forward that the relative immunity of chronic carriers to epidemic meningitis may be due to the presence of specific antibodies in the blood stream.


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