Preparation of a working seed lot of BCG and quality control by PCR genotyping
The bacillus Calmette-Guérin (BCG) was obtained in 1920 after successive passages leading to the attenuation of a Mycobacterium bovis strain. For the following 40 years, BCG had been replicated, resulting in substrains with genotypic and phenotypic differences. Several genomic studies have compared...
| Main Authors: | , , |
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| Format: | Artículo |
| Language: | Inglés |
| Published: |
Asociación Argentina de Microbiología
2019
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| Online Access: | http://www.scielo.org.ar/pdf/ram/v42n1/v42n1a02.pdf http://hdl.handle.net/20.500.12123/6362 |
| _version_ | 1855483797203582976 |
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| author | Trovero, A. Argüelles, Claudia Cataldi, Angel Adrian |
| author_browse | Argüelles, Claudia Cataldi, Angel Adrian Trovero, A. |
| author_facet | Trovero, A. Argüelles, Claudia Cataldi, Angel Adrian |
| author_sort | Trovero, A. |
| collection | INTA Digital |
| description | The bacillus Calmette-Guérin (BCG) was obtained in 1920 after successive passages leading to the attenuation of a Mycobacterium bovis strain. For the following 40 years, BCG had been replicated, resulting in substrains with genotypic and phenotypic differences. Several genomic studies have compared two BCG strains, M. bovis and Mycobacterium tuberculosis, and observed that deleted regions in the different strains could be related to differences in antigenic properties. In this work, a working seed lot was obtained from a lyophilized secondary seed lot from the BCG Pasteur strain 1173 P2 and genetically characterized. The genome was analyzed by PCR directed to five regions (RD1, RD2, RD14, RD15, DU2), using the seed lot and different available strains as templates. No genetic differences were found in the fragments studied as compared to the Pasteur strain. A total of 20 passages were carried out and no differences were found in the size of the fragments amplified by PCR. In conclusion, this method allows to control a working seed lot genotypically and to assess the stability of the BCG genome. |
| format | Artículo |
| id | INTA6362 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2019 |
| publishDateRange | 2019 |
| publishDateSort | 2019 |
| publisher | Asociación Argentina de Microbiología |
| publisherStr | Asociación Argentina de Microbiología |
| record_format | dspace |
| spelling | INTA63622019-11-21T17:04:38Z Preparation of a working seed lot of BCG and quality control by PCR genotyping Trovero, A. Argüelles, Claudia Cataldi, Angel Adrian Genomas PCR Mycobacterium Bovis Control de Calidad Genética Genomes Quality Controls Genetics Bacillus Calmette-Guérin Bacilo Calmette-Guérin The bacillus Calmette-Guérin (BCG) was obtained in 1920 after successive passages leading to the attenuation of a Mycobacterium bovis strain. For the following 40 years, BCG had been replicated, resulting in substrains with genotypic and phenotypic differences. Several genomic studies have compared two BCG strains, M. bovis and Mycobacterium tuberculosis, and observed that deleted regions in the different strains could be related to differences in antigenic properties. In this work, a working seed lot was obtained from a lyophilized secondary seed lot from the BCG Pasteur strain 1173 P2 and genetically characterized. The genome was analyzed by PCR directed to five regions (RD1, RD2, RD14, RD15, DU2), using the seed lot and different available strains as templates. No genetic differences were found in the fragments studied as compared to the Pasteur strain. A total of 20 passages were carried out and no differences were found in the size of the fragments amplified by PCR. In conclusion, this method allows to control a working seed lot genotypically and to assess the stability of the BCG genome. El bacilo de Calmette-Guérin (BCG) se obtuvo en 1920, después de sucesivos pasajes que llevaron a la atenuación de una cepa de Mycobacterium bovis. A lo largo de los 40 años subsiguientes la cepa BCG fue replicada y surgieron subcepas con diferencias fenotípicas y genotípicas. Se realizaron varios estudios de comparación genómica de diferentes cepas de BCG, M. bovis y Mycobacterium tuberculosis, y se observó que las deleciones de regiones en las diferentes cepas podrían estar relacionadas con diferencias en las propiedades antigénicas. En este trabajo se describe la preparación y caracterización genética de un lote semilla de trabajo obtenido a partir de un lote semilla secundaria liofilizado de la cepa BCG Pasteur 1173 P2. Se analizaron por PCR cinco regiones (RD1, RD2, RD14, RD15, DU2) en el lote semilla de trabajo utilizando como control las diferentes cepas disponibles. No se hallaron diferencias genéticas en los fragmentos estudiados al comparar el lote semilla de trabajo con la cepa BCG Pasteur 1173 P2. Asimismo, se efectuaron hasta 20 pasajes y no se encontraron diferencias en el tamaño de los fragmentos amplificados por PCR. En conclusión, se ha puesto a punto un método que permite controlar el genotipo de un lote semilla de trabajo y evaluar la estabilidad del genoma del BCG. Instituto de Biotecnología Fil: Trovero, A. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Producción de Biológicos; Argentina. Fil: Argüelles, Claudia. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Producción de Biológicos; Argentina. Fil: Cataldi, Angel Adrian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina 2019-11-21T17:02:38Z 2019-11-21T17:02:38Z 2010-04 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://www.scielo.org.ar/pdf/ram/v42n1/v42n1a02.pdf http://hdl.handle.net/20.500.12123/6362 0325-7541 1851-7617 eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Asociación Argentina de Microbiología Revista Argentina de Microbiología 42 (1) : 4–10. (Ene.-Abr. 2010) |
| spellingShingle | Genomas PCR Mycobacterium Bovis Control de Calidad Genética Genomes Quality Controls Genetics Bacillus Calmette-Guérin Bacilo Calmette-Guérin Trovero, A. Argüelles, Claudia Cataldi, Angel Adrian Preparation of a working seed lot of BCG and quality control by PCR genotyping |
| title | Preparation of a working seed lot of BCG and quality control by PCR genotyping |
| title_full | Preparation of a working seed lot of BCG and quality control by PCR genotyping |
| title_fullStr | Preparation of a working seed lot of BCG and quality control by PCR genotyping |
| title_full_unstemmed | Preparation of a working seed lot of BCG and quality control by PCR genotyping |
| title_short | Preparation of a working seed lot of BCG and quality control by PCR genotyping |
| title_sort | preparation of a working seed lot of bcg and quality control by pcr genotyping |
| topic | Genomas PCR Mycobacterium Bovis Control de Calidad Genética Genomes Quality Controls Genetics Bacillus Calmette-Guérin Bacilo Calmette-Guérin |
| url | http://www.scielo.org.ar/pdf/ram/v42n1/v42n1a02.pdf http://hdl.handle.net/20.500.12123/6362 |
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