Validation of an ELISA-type immune-enzymatic test to quantify levels of diphtheria antitoxin in human serum
<p><strong>Introduction:</strong> Diphtheria still persists in many countries. In Cuba, studies conducted in different age groups have demonstrated that there are non-protective levels of diphtheria antitoxin in the population, so it is necessary to have methods that allow the sero...
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| Format: | Article |
| Language: | English |
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Universidad de Ciencias Médicas de La Habana
2018-09-01
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| Series: | Revista Habanera de Ciencias Médicas |
| Online Access: | http://www.revhabanera.sld.cu/index.php/rhab/article/view/2113 |
| _version_ | 1828794665964404736 |
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| author | Cira Virgen Rodríguez Pelier Yaíma Zúñiga Rosales Bárbara Torres Rives Minerva Matarán Valdéz |
| author_facet | Cira Virgen Rodríguez Pelier Yaíma Zúñiga Rosales Bárbara Torres Rives Minerva Matarán Valdéz |
| author_sort | Cira Virgen Rodríguez Pelier |
| collection | DOAJ |
| description | <p><strong>Introduction:</strong> Diphtheria still persists in many countries. In Cuba, studies conducted in different age groups have demonstrated that there are non-protective levels of diphtheria antitoxin in the population, so it is necessary to have methods that allow the serologic survey of population immunity. The quantification of antibodies against vaccine antigens such as diphtheria toxin is also a useful, rapid and economic method to evaluate the immune response.</p><p><strong>Objective:</strong> To validate an ELISA-type immune-enzymatic test to quantify the levels of diphtheria antitoxin in human serum.</p><p><strong>Material and Method:</strong> An experimental study of technological development was carried out in the Immunology Laboratory of the National Medical Genetics Center, Havana, Cuba. The optimal values of the variables that influence on the result of the indirect heterogeneous immune-enzymatic test for the quantification of diphtheria antitoxin were determined. The calibration curve obtained was evaluated against the WHO standard (Diphtheria Antitoxin Human Serum 00/496).<strong> </strong>The analytical validation of the standardized method was performed.</p><p><strong>Results:</strong> The intra-assay and inter-assay coefficients of variation were less than 10% and 20%, respectively. Recovery values between 90 and 110% were found in accuracy and selectivity. The parallelism between the standard curve and the samples studied showed a coefficient of variation lower or equal to 10%. The limit of quantification was 0,015 IU/mL and the one of detection was 0,0039 IU/mL. </p><p><strong>Conclusions:</strong> The result obtained in the precision, accuracy and selectivity of the ELISA-type immune-enzymatic test developed and validated in the National Medical Genetics Center demonstrated that it can be used in the clinical practice to quantify the values of diphtheria antitoxin in human serum.</p><p><strong>Keywords:</strong> diphtheria antitoxin, immune-enzymatic test, ELISA, validation.</p> |
| first_indexed | 2024-12-12T03:50:15Z |
| format | Article |
| id | doaj.art-3f0c0f270dc040d8a6ae486a02707487 |
| institution | Directory Open Access Journal |
| issn | 1729-519X |
| language | English |
| last_indexed | 2024-12-12T03:50:15Z |
| publishDate | 2018-09-01 |
| publisher | Universidad de Ciencias Médicas de La Habana |
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| series | Revista Habanera de Ciencias Médicas |
| spelling | doaj.art-3f0c0f270dc040d8a6ae486a027074872022-12-22T00:39:25ZengUniversidad de Ciencias Médicas de La HabanaRevista Habanera de Ciencias Médicas1729-519X2018-09-011745275391388Validation of an ELISA-type immune-enzymatic test to quantify levels of diphtheria antitoxin in human serumCira Virgen Rodríguez Pelier0Yaíma Zúñiga Rosales1Bárbara Torres Rives2Minerva Matarán Valdéz3Universidad de Ciencias Médicas de La Habana. Centro Nacional de Genética Médica. La Habana.Universidad de Ciencias Médicas de La Habana. Centro Nacional de Genética Médica. La Habana.Universidad de Ciencias Médicas de La Habana. Centro Nacional de Genética Médica. La Habana.Centro Nacional de Genética Médica. La Habana.<p><strong>Introduction:</strong> Diphtheria still persists in many countries. In Cuba, studies conducted in different age groups have demonstrated that there are non-protective levels of diphtheria antitoxin in the population, so it is necessary to have methods that allow the serologic survey of population immunity. The quantification of antibodies against vaccine antigens such as diphtheria toxin is also a useful, rapid and economic method to evaluate the immune response.</p><p><strong>Objective:</strong> To validate an ELISA-type immune-enzymatic test to quantify the levels of diphtheria antitoxin in human serum.</p><p><strong>Material and Method:</strong> An experimental study of technological development was carried out in the Immunology Laboratory of the National Medical Genetics Center, Havana, Cuba. The optimal values of the variables that influence on the result of the indirect heterogeneous immune-enzymatic test for the quantification of diphtheria antitoxin were determined. The calibration curve obtained was evaluated against the WHO standard (Diphtheria Antitoxin Human Serum 00/496).<strong> </strong>The analytical validation of the standardized method was performed.</p><p><strong>Results:</strong> The intra-assay and inter-assay coefficients of variation were less than 10% and 20%, respectively. Recovery values between 90 and 110% were found in accuracy and selectivity. The parallelism between the standard curve and the samples studied showed a coefficient of variation lower or equal to 10%. The limit of quantification was 0,015 IU/mL and the one of detection was 0,0039 IU/mL. </p><p><strong>Conclusions:</strong> The result obtained in the precision, accuracy and selectivity of the ELISA-type immune-enzymatic test developed and validated in the National Medical Genetics Center demonstrated that it can be used in the clinical practice to quantify the values of diphtheria antitoxin in human serum.</p><p><strong>Keywords:</strong> diphtheria antitoxin, immune-enzymatic test, ELISA, validation.</p>http://www.revhabanera.sld.cu/index.php/rhab/article/view/2113 |
| spellingShingle | Cira Virgen Rodríguez Pelier Yaíma Zúñiga Rosales Bárbara Torres Rives Minerva Matarán Valdéz Validation of an ELISA-type immune-enzymatic test to quantify levels of diphtheria antitoxin in human serum Revista Habanera de Ciencias Médicas |
| title | Validation of an ELISA-type immune-enzymatic test to quantify levels of diphtheria antitoxin in human serum |
| title_full | Validation of an ELISA-type immune-enzymatic test to quantify levels of diphtheria antitoxin in human serum |
| title_fullStr | Validation of an ELISA-type immune-enzymatic test to quantify levels of diphtheria antitoxin in human serum |
| title_full_unstemmed | Validation of an ELISA-type immune-enzymatic test to quantify levels of diphtheria antitoxin in human serum |
| title_short | Validation of an ELISA-type immune-enzymatic test to quantify levels of diphtheria antitoxin in human serum |
| title_sort | validation of an elisa type immune enzymatic test to quantify levels of diphtheria antitoxin in human serum |
| url | http://www.revhabanera.sld.cu/index.php/rhab/article/view/2113 |
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