scholarly journals Development of Elisa Test-Systems and Immune-Chromatographic Reagent Panel Designed for the Detection of Staphylococcal Enterotoxins of A and B Types

2021 ◽  
pp. 94-99
Author(s):  
A. V. Eremkin ◽  
S. S. Ipatov ◽  
G. V. Kuklina ◽  
D. V. Pechenkin ◽  
A. A. Kytmanov ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Elisa Test ◽  
Food Samples ◽  
Central Research Institute ◽  
Exchange Chromatography ◽  
Lateral Flow ◽  
Hybridoma Cells ◽  
Central Research ◽  
Staphylococcal Enterotoxins ◽  
The Russian Federation

Objective of the study was the development of experimental ELISA tests and lateral flow immunoassays for detection of staphylococcal enterotoxins, A and B types.Materials and methods. Hybridomas, producing monoclonal antibodies against staphylococcal enterotoxins A and B from State Collection of the Affiliated Branch of the “48th Central Research Institute” of the Ministry of Defense of the Russian Federation, BALB/c mice and staphylococcal enterotoxins A and B were used in the research. Hybridoma cells were incubated in culture flasks and in the peritoneal cavity of BALB/c mice. Monoclonal antibodies were isolated from ascitic fluids through precipitation with saturated ammonium sulfate subsequently purified using ion-exchange chromatography. Obtained preparations of monoclonal antibodies were used for the construction of ELISA tests and immune-chromatographic reagent panels for the detection of staphylococcal enterotoxins A and B. Specific components of ELISA tests were lyophilized in protective media.Results and discussion. ELISA tests and lateral flow immunoassays which allow for detecting staphylococcal enterotoxins A and B at concentrations of 0.5 ng/ml and higher, including in food samples, have been constructed. 

Development of enzyme immunoassay for detecting I and II types of shiga-like toxins

2021 ◽  
Vol 29 (5) ◽  
pp. 43-48
Author(s):  
Galina Viktorovna Kuklina ◽  
Denis Valerievich Pechenkin ◽  
Sergei Sergeevich Ipatov ◽  
Andrei Valentinovich Eremkin ◽  
Aleksei Aleksandrovich Kytmanov ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Enzyme Immunoassay ◽  
Solid Phase ◽  
Sandwich Elisa ◽  
Central Research Institute ◽  
Exchange Chromatography ◽  
Hybridoma Cells ◽  
Federal State ◽  
Central Research ◽  
Freeze Dried

Introduction. The aim of the work was development of enzyme immunoassay for detecting I and II types of shiga-like toxins and assessment of it diagnostic properties. Materials and methods. For the research, we used hybridomas producing monoclonal antibodies to shiga-like toxins of types I and II, obtained at the branch of the Federal State Budgetary Institution “48 Central Research Institute” of the Ministry of Defense of Russian Federation (Kirov); BALB/c mice; shiga-like toxins of types I and II. Hybridoma cells were cultured in culture flasks and in the peritoneal cavity of BALB/c mice. Monoclonal antibodies were isolated from ascitic fluids by precipitation with a saturated solution of ammonium sulfate, followed by purification by ion exchange chromatography. The obtained preparations of monoclonal antibodies were used to develop enzyme immunoassay for the detection of shiga-like toxins of types I and II. Specific components of enzyme immunoassay were freeze-dried in a protective environment. Results. As a result of research, preparative quantities of monoclonal antibodies against I and II types of shiga-like toxins were obtained and purified; selection of monoclonal antibodies for sorption on the solid phase and for the synthesis of immunoperoxidase conjugates was carried out. Conclusion. experimental enzyme immunoassay allowing to identify 1 ng/ml I and II types of shiga-like toxins in «sandwich»-ELISA was developed.

scholarly journals Development of Enzyme-Immunoassay for Bacillus anthracis Detection

2018 ◽  
pp. 78-82
Author(s):  
D. V. Pechenkin ◽  
O. O. Fomenkov ◽  
A. V. Eremkin ◽  
G. D. Elagin ◽  
G. V. Kuklina ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Bacillus Anthracis ◽  
Enzyme Immunoassay ◽  
Specific Activity ◽  
Hybrid Cell ◽  
Sandwich Elisa ◽  
Central Research Institute ◽  
Exchange Chromatography ◽  
Central Research ◽  
Test Kit

Objective of the study was to develop enzyme-immunoassay test-kit for the detection of Bacillus anthracis spores. Materials and methods. Microbial cultures from the State Collection of Microorganisms at the premises of Affiliated Branch of the «48th Central Research Institute» of the Ministry of Defense of the Russian Federation and BALB/c mice were used in the research. Hybridization of B-lymphocytes with SP2/0-Ag14 myeloma cells was performed according to G. Kohler and C. Milstein procedure in De St. Fazekas and P. Scheidegger modification. Hybridomas were cultured in the peritoneal cavity of BALB/c mice. Ascitic fluids were isolated from mice, precipitated with ammonium sulfate and purified by means of ion-exchange chromatography for preparation of monoclonal antibodies. Specific activity of hybridoma’s supernatants, ascitic fluids, purified monoclonal antibodies was studied by «sandwich» ELISA. Specific components of test-kit were lyophilized in suitable cryoprotective medium. Results and conclusions.We have obtained new hybrid cell lines producing specific monoclonal antibodies against Bacillus anthracisspore antigens and ascitic fluids from which immunoglobulins were isolated. Optimum combinations of monoclonal antibodies as a sensitizer and a component of immunoperoxidase conjugates have been selected. Monoclonal antibodies 272E10G1-272F7A10 provide the highest sensitivity of ELISA for the detection of anthrax microbe spore antigens. Our enzyme-immunoassay test allows for identification of Bacillus anthracis spores in concentrations up to 5,0·105 spores per milliliter. No cross reaction with closely related saprophytes and other heterologous microorganisms in concentrations of 1,0·108 CFU per milliliter is observed.

scholarly journals Development of Immuno-Enzymatic Monoclonal Tests-Systems for the Detection of Glanders and Melioidosis Agents

2018 ◽  
pp. 60-65
Author(s):  
A. A. Kytmanov ◽  
G. D. Elagin ◽  
G. V. Kuklina ◽  
D. V. Pechenkin ◽  
O. O. Fomenkov ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Hybrid Cell ◽  
Cross Reactivity ◽  
Central Research Institute ◽  
Hybridoma Cells ◽  
Central Research ◽  
Test Systems ◽  
Test Kit

Objective of the study was the development of immune-enzymatic monoclonal test-kit for detecting glanders and melioidosis agents. Materials and methods. We used microbial cultures and hybrid cell lines obtained from the collection of the «48th Central Research Institute» of the Ministry of Defense of the Russian Federation. Hybridoma cells were incubated in the peritoneal cavity of BALB/c mice. Preparations of glanders and melioidosis monoclonal antibodies were isolated from the ascetic fluids through precipitation with ammonium sulfate and purification by means of ion-exchange chromatography. Specific components of the test-kits were subjected to freeze drying in corresponding protective media. Study of diagnostic properties of the developed test systems was performed using ELISA. Results and conclusions. We have obtained preparations of monoclonal antibodies in vivo, as well as isolated and purified immunoglobulins from ascetic fluids. We also selected the pairs of monoclonal antibodies for manufacturing specific components. Experimental series of immune-enzymatic monoclonal test-systems allowing for specific detection of glanders and melioidosis causative agents in concentrations ranging from 0.5·106 CFU/ml and higher were made. The absence of cross-reactivity with closely related saprophytes and heterologous microorganisms in concentrations of 1,0·108 CFU/ml was shown. Demonstrated was the possibility in principle to differentiate between Burkholderia malleiand Burkholderia pseudomallei using ELISA. Test systems are promising for follow up state registration as medical products for in vitro diagnostics.

scholarly journals INCIDENCE OF PSORIASIS DEPENDING ON THE REGION OF THE RUSSIAN FEDERATION

2018 ◽  
Vol 21 (2) ◽  
pp. 85-89
Author(s):  
Zh. V Gudinova ◽  
V. A Okhlopkov ◽  
Ekaterina Ivanovna Poleshchuk ◽  
G. N Zhernakova
Keyword(s):  
Cluster Analysis ◽  
Russian Federation ◽  
Komi Republic ◽  
Central Research Institute ◽  
Central Research ◽  
Incidence Data ◽  
The North ◽  
Class 1 ◽  
The Russian Federation ◽  
Health Organization

The article presents the results of a study on assessment the differences of psoriasis incidence in adults in regions of the Russian Federation. The indicators of general and primary incidence of psoriasis were used as the material for a study according to the data from Central Research Institute of Health Organization and Informatics of the Ministry of Public Health of the Russian Federation for the period 2009-2013. The comparative results of psoriasis prevalence among adults population in various regions in Russia were presented. Higher incidence rate was detected in the northern and northeastern regions of the North-Western, Volga, Ural, and Far Eastern regions. The results of classification for the regions of the Russian Federation according to the data of primary and general morbidity were presented. In order to establish the peculiarities of pathology formation and assess the quality of information in the regions of the Russian Federation, a multidimensional classification of the incidence of psoriasis was carried out using the cluster analysis method, using the comparison of prevalence (general morbidity) with primary incidence data for the period 2009-2011. As a result of the cluster analysis, four classes of the country regions are distinguished, the differences between which are statistically significant for all classification characteristics. The average indices of both primary incidence and prevalence of psoriasis are registered in regions of Class 1 (27 subjects of the Russian Federation). The minimum indices of primary and general incidence of psoriasis were noted in class 2 (24 subjects of the Russian Federation), which comprised regions predominantly of the Central, Southern, Siberian FD. The highest rates of primary incidence and prevalence of psoriasis were found in the group of regions that made up Class 4 (14 subjects of the Russian Federation), in a third of the regions of this class - Orel and Sverdlovsk regions, Udmurt Republic, Komi Republic and Sakha (Yakutia), data, unsatisfactory dispensary observation of patients with psoriasis. A certain decrease in the availability of specialized dermatological care can be expected in Class 3 regions, where the primary incidence is low, and the prevalence is above the average (18 subjects of the Russian Federation).

scholarly journals The Level of Meningococcal Carriage and Genotyping of N. meningitidis Strains in the Group of Labor Migrants

2020 ◽  
Vol 19 (5) ◽  
pp. 25-33
Author(s):  
M. A. Koroleva ◽  
M. I. Gritsay ◽  
K. O. Mironov ◽  
N. N. Fomkina ◽  
I. S. Koroleva ◽  
...  
Keyword(s):  
Local Level ◽  
Federal Budget ◽  
Central Research Institute ◽  
Dynamic Monitoring ◽  
Meningococcal Infection ◽  
Central Research ◽  
Labor Migrants ◽  
Country Level ◽  
The Russian Federation ◽  
Biological Methods

Relevance. Population migration can play a crucial role in the spread of invasive strains of meningococcus, initiating outbreaks of meningococcal infection, and changing the incidence at the local level.Aim. To assess the prevalence of meningococcal carriage among migrants arriving in Moscow and to characterize the antigenic and genetic properties of carrier strains of meningococcus.Materials and methods. The study was conducted in March 2020 at the bases of the Multifunctional Migration Center of Moscow and the Federal Budget Institution of Science «Central Research Institute of Epidemiology». Samples of nasopharyngeal mucus were collected from 352 people. Nasopharyngeal strains of meningococcus were identified and identified using microbiological, serological, and molecular biological methods.Results. The overall level of the carriage was 5.7%. Of the twenty selected strains, 10 have a serogroup defined: Y – 5 strains, W - 3, A, and B – 1 each. The obtained genetic and antigenic characteristics do not allow talking about the import into the RF of representatives of known hypervirulent clonal complexes. In this study, strains were identified that are part of the clonal complex ST-175 complex, which has not been previously described in the Russian Federation.Conclusion. It seems promising to continue the dynamic monitoring of carriage of meningococcus in various groups, including among people entering the country to obtain a migration patent, as well as identifying risk factors for acquiring carriage. The data obtained will supplement current information on the incidence of the generalized form of meningococcal infection and will be crucial for determining the epidemiology at the country level, the population groups responsible for the transmission of the disease, and the need for targeted vaccination.

Evaluation of the Efficiency of Protein A Affinity Chromatography to Purify a Monoclonal Antibody for Cancer Treatment and its Purity Analysis

2020 ◽  
Vol 7 (2) ◽  
pp. 121-133
Author(s):  
Ayesha Akhtar ◽  
Shivakumar Arumugam ◽  
Shoaib Alam
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Affinity Chromatography ◽  
Protein A ◽  
Exchange Chromatography ◽  
Size Exclusion ◽  
High Yield ◽  
Staphylococcal Protein A ◽  
Purification Step ◽  
Purity Analysis

Background:: Protein A affinity chromatography is often employed as the most crucial purification step for monoclonal antibodies to achieve high yield with purity and throughput requirements. Introduction:: Protein A, also known as Staphylococcal protein A (SPA) is found in the cell wall of the bacteria staphylococcus aureus. It is one of the first discovered immunoglobulin binding molecules and has been extensively studied since the past few decades. The efficiency of Protein A affinity chromatography to purify a recombinant monoclonal antibody in a cell culture sample has been evaluated, which removes 99.0% of feed stream impurities. Materials and Method:: We have systematically evaluated the purification performance by using a battery of analytical methods SDS-PAGE (non-reduced and reduced sample), Cation Exchange Chromatography (CEX), Size-exclusion chromatography (SEC), and Reversed phased-Reduced Chromatography for a CHO-derived monoclonal antibody. Results and Discussion:: The analytical test was conducted to determine the impurity parameter, Host Cell Contaminating Proteins (HCP). It was evaluated to be 0.015ng/ml after the purification step; while initially, it was found to be 24.431ng/ml. Conclusion:: The tests showed a distinct decrease in the level of different impurities after the chromatography step. It can be concluded that Protein A chromatography is an efficient step in the purification of monoclonal antibodies.

scholarly journals Regional differences in general practitioners’ behaviours regarding influenza vaccination: a cross-sectional study

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Jonathan Arlt ◽  
Kristina Flaegel ◽  
Katja Goetz ◽  
Jost Steinhaeuser
Keyword(s):  
Influenza Vaccination ◽  
General Practitioners ◽  
Personal Characteristics ◽  
Cross Sectional Study ◽  
Central Research Institute ◽  
World Health ◽  
Central Research ◽  
Cross Sectional ◽  
Vaccination Rates ◽  
Health Organization

Abstract Background The World Health Organization recommends vaccination rates of 75% against seasonal influenza for patients over 65 years old. In the 2013/2014 season, the German vaccination rates ranged between 14 and 65%. This study aimed to compare the attitudes, personal characteristics and vaccination behaviours of general practitioners (GPs) in regions with high and low vaccination rates in Germany. Methods In May 2016, a questionnaire was sent to 1594 GPs practising in 16 districts with the highest and the lowest vaccination rates in Western and Eastern Germany as described by the Central Research Institute of Ambulatory Health Care in Germany for the 2013/2014 season. Descriptive statistics and multiple regression analyses were computed to identify potential factors associated with high vaccination rates. Results A total response rate of 32% (515/1594 participants) was observed in the study. GPs reported their attitudes towards vaccination in general and vaccination against influenza as mostly ‘very positive’ (80%, n = 352 and 65%, n = 288, respectively). GPs practising in regions with low vaccination rates reported their attitudes towards vaccinations in general (p = 0.004) and towards influenza vaccination (p = 0.001) more negatively than their colleagues from regions with high vaccination rates. Multiple logistic regression identified an increasing influence of year-dependent changing efficiency on GPs’ influenza rates as the strongest factor for predicting GPs from highly vaccinating regions (OR = 4.31 [1.12–16.60]), followed by the patient’s vaccination refusal despite GP advice due to already receiving a vaccination from another physician (OR = 3.20 [1.89–5.43]) and vaccination information gathering through medical colleagues (OR = 2.26 [1.19–4.29]). Conclusions The results of this study suggest a correlation between GPs’ attitudes and regional vaccination rates. Beneath GPs’ individual attitudes, the regional attitude patterns of patients, colleagues and medical assistants surrounding those GPs seem decisive and should be integrated into future campaigns to increase vaccination rates at a regional level.

scholarly journals On the experimental production of congenital goitre

1916 ◽  
Vol 89 (616) ◽  
pp. 322-327
Keyword(s):  
Central Research Institute ◽  
Central Research ◽  
Experimental Production ◽  
The Past ◽  
White Rats ◽  
Military Duty ◽  
Active Service ◽  
Large Animals ◽  
Research Institute

The experiment herein described was undertaken at the Central Research Institute, Kasauli, India. It commenced on September 6, 1913, and terminated, owing to my recall to military duty for active service, on December 24, 1914. Having been on service for the past 18 months I have not hitherto had an opportunity to report it. Object of the Experiment . Its object was to determine the cause of congenital goitre and the conditions under which it developed in large animals, and to confirm and amplify the results I had obtained by previous experimentation on white rats.

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