Obtaining immunodiagnostic preparations from <i>Toxocara canis</i> antigens for serodiagnostics of toxocariasis in human
Toxocariasis is a human helminthic invasion that has a wide range of hosts with epizootic distribution. The populational seropositivity in countries with a temperate climate comprises about 37%, whereas in regions with tropical climate up to 92%. Almost all age groups of the population are at risk...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | Russian |
| Published: |
Sankt-Peterburg : NIIÈM imeni Pastera
2022-05-01
|
| Series: | Инфекция и иммунитет |
| Subjects: | |
| Online Access: | https://iimmun.ru/iimm/article/viewFile/1804/1448 |
| _version_ | 1811186707124453376 |
|---|---|
| author | Irina V. Kanina A. I. Novak M. D. Novak O. V. Evdokimova |
| author_facet | Irina V. Kanina A. I. Novak M. D. Novak O. V. Evdokimova |
| author_sort | Irina V. Kanina |
| collection | DOAJ |
| description | Toxocariasis is a human helminthic invasion that has a wide range of hosts with epizootic distribution. The populational seropositivity in countries with a temperate climate comprises about 37%, whereas in regions with tropical climate up to 92%. Almost all age groups of the population are at risk of invasion by toxocars. The immunological method currently retains diagnostic significance for toxocariasis, because the reaction of immune system against helminths is accompanied by developing sensitization, and techniques as well as methods of collecting clinical material to identify the migratory form of toxocarosis in humans are time-consuming and invasive. The purpose of this study is to develop an immunobiological preparation based on excretory-secretory antigens from Toxocara canis larvae for serodiagnostics of larva migrans syndrome in human by using enzyme-linked immunoassay (ELISA). Antigens were obtained from Toxocara canis larvae by culturing its eggs isolated from the uterus of female nematodes in the glutamine-supplemented medium. The preparation was purified from ballast substances by centrifugation at 8000 g for 20 minutes followed by filtration through microfiltration membrane with a pore diameter of 0,050,15 microns, type MFAS-P-1 (manufactured by CJSC STC Vladipor). Blood serum of student volunteers from countries with high incidence rate of toxocariasis were tested to detect T. canis-specific antibodies. ELISA with commercial antigens Toxocara-IgG-ELISA-BEST kit was used for selection of seropositive and seronegative sera also used as a control. Diagnostic antigenic preparations with different protein concentrations were prepared to examine seropositive sera for determining optimal dose of the antigenic drugs. Results. ELISA with standard test kit allowed to detect Toxocara-IgG antibodies in 20 sera (8%) from 250 samples, what indicates about antigenic stimulation of the immune system by helminths and suspected former disease. Analyzing experimental samples allowed to find the number of seropositive data correlated with antigenic protein concentration in preparations: at a concentration of 1,96 g/ml 5,2% of positive results were detected, 1,71 g/ml 3.2%, 0.33 g/ml 0.8% (Pearsons correlation coefficient 0,94). The number of positive sera detected with commercial antigen in ELISA kit was identical to that of positive sera detected by an experimental antigen sample with a protein concentration of 2,49 g/ml. Thus, the immunodiagnostic preparations obtained by the experimental method based on the excretory-secretory antigens from Toxocara canis are characterized by high sensitivity and can be used to detect Toxocara-IgG antibodies in ELISA. The antigen preparation protein concentration of at least 2.49 g/ml is optimal and correlates whit sensitivity of the commercial antigen of the standard ELISA. |
| first_indexed | 2024-04-11T13:50:02Z |
| format | Article |
| id | doaj.art-d4c4400e43fc4813a15332cdffa16ab0 |
| institution | Directory Open Access Journal |
| issn | 2220-7619 2313-7398 |
| language | Russian |
| last_indexed | 2024-04-11T13:50:02Z |
| publishDate | 2022-05-01 |
| publisher | Sankt-Peterburg : NIIÈM imeni Pastera |
| record_format | Article |
| series | Инфекция и иммунитет |
| spelling | doaj.art-d4c4400e43fc4813a15332cdffa16ab02022-12-22T04:20:41ZrusSankt-Peterburg : NIIÈM imeni PasteraИнфекция и иммунитет2220-76192313-73982022-05-0112239139610.15789/2220-7619-OID-18041150Obtaining immunodiagnostic preparations from <i>Toxocara canis</i> antigens for serodiagnostics of toxocariasis in humanIrina V. Kanina0https://orcid.org/0000-0002-8419-1697A. I. Novak1https://orcid.org/0000-0003-0345-7316M. D. Novak2https://orcid.org/0000-0002-1421-0039O. V. Evdokimova3https://orcid.org/0000-0002-5035-7302Ryazan State Medical University named after academician I.P. PavlovRyazan State Medical University named after academician I.P. PavlovRyazan State Medical University named after academician I.P. PavlovRyazan State Medical University named after academician I.P. PavlovToxocariasis is a human helminthic invasion that has a wide range of hosts with epizootic distribution. The populational seropositivity in countries with a temperate climate comprises about 37%, whereas in regions with tropical climate up to 92%. Almost all age groups of the population are at risk of invasion by toxocars. The immunological method currently retains diagnostic significance for toxocariasis, because the reaction of immune system against helminths is accompanied by developing sensitization, and techniques as well as methods of collecting clinical material to identify the migratory form of toxocarosis in humans are time-consuming and invasive. The purpose of this study is to develop an immunobiological preparation based on excretory-secretory antigens from Toxocara canis larvae for serodiagnostics of larva migrans syndrome in human by using enzyme-linked immunoassay (ELISA). Antigens were obtained from Toxocara canis larvae by culturing its eggs isolated from the uterus of female nematodes in the glutamine-supplemented medium. The preparation was purified from ballast substances by centrifugation at 8000 g for 20 minutes followed by filtration through microfiltration membrane with a pore diameter of 0,050,15 microns, type MFAS-P-1 (manufactured by CJSC STC Vladipor). Blood serum of student volunteers from countries with high incidence rate of toxocariasis were tested to detect T. canis-specific antibodies. ELISA with commercial antigens Toxocara-IgG-ELISA-BEST kit was used for selection of seropositive and seronegative sera also used as a control. Diagnostic antigenic preparations with different protein concentrations were prepared to examine seropositive sera for determining optimal dose of the antigenic drugs. Results. ELISA with standard test kit allowed to detect Toxocara-IgG antibodies in 20 sera (8%) from 250 samples, what indicates about antigenic stimulation of the immune system by helminths and suspected former disease. Analyzing experimental samples allowed to find the number of seropositive data correlated with antigenic protein concentration in preparations: at a concentration of 1,96 g/ml 5,2% of positive results were detected, 1,71 g/ml 3.2%, 0.33 g/ml 0.8% (Pearsons correlation coefficient 0,94). The number of positive sera detected with commercial antigen in ELISA kit was identical to that of positive sera detected by an experimental antigen sample with a protein concentration of 2,49 g/ml. Thus, the immunodiagnostic preparations obtained by the experimental method based on the excretory-secretory antigens from Toxocara canis are characterized by high sensitivity and can be used to detect Toxocara-IgG antibodies in ELISA. The antigen preparation protein concentration of at least 2.49 g/ml is optimal and correlates whit sensitivity of the commercial antigen of the standard ELISA.https://iimmun.ru/iimm/article/viewFile/1804/1448toxocariasisimmunodiagnosticsexcretory-secretory antigenlinked immunosorbent assaylarvaediagnosticum |
| spellingShingle | Irina V. Kanina A. I. Novak M. D. Novak O. V. Evdokimova Obtaining immunodiagnostic preparations from <i>Toxocara canis</i> antigens for serodiagnostics of toxocariasis in human Инфекция и иммунитет toxocariasis immunodiagnostics excretory-secretory antigen linked immunosorbent assay larvae diagnosticum |
| title | Obtaining immunodiagnostic preparations from <i>Toxocara canis</i> antigens for serodiagnostics of toxocariasis in human |
| title_full | Obtaining immunodiagnostic preparations from <i>Toxocara canis</i> antigens for serodiagnostics of toxocariasis in human |
| title_fullStr | Obtaining immunodiagnostic preparations from <i>Toxocara canis</i> antigens for serodiagnostics of toxocariasis in human |
| title_full_unstemmed | Obtaining immunodiagnostic preparations from <i>Toxocara canis</i> antigens for serodiagnostics of toxocariasis in human |
| title_short | Obtaining immunodiagnostic preparations from <i>Toxocara canis</i> antigens for serodiagnostics of toxocariasis in human |
| title_sort | obtaining immunodiagnostic preparations from i toxocara canis i antigens for serodiagnostics of toxocariasis in human |
| topic | toxocariasis immunodiagnostics excretory-secretory antigen linked immunosorbent assay larvae diagnosticum |
| url | https://iimmun.ru/iimm/article/viewFile/1804/1448 |
| work_keys_str_mv | AT irinavkanina obtainingimmunodiagnosticpreparationsfromitoxocaracanisiantigensforserodiagnosticsoftoxocariasisinhuman AT ainovak obtainingimmunodiagnosticpreparationsfromitoxocaracanisiantigensforserodiagnosticsoftoxocariasisinhuman AT mdnovak obtainingimmunodiagnosticpreparationsfromitoxocaracanisiantigensforserodiagnosticsoftoxocariasisinhuman AT ovevdokimova obtainingimmunodiagnosticpreparationsfromitoxocaracanisiantigensforserodiagnosticsoftoxocariasisinhuman |