Purificación del virus del síndrome de la hoja amarilla de la caña de azúcar (Sugarcane yellow leaf luteovirus ScYLV) y producción de antisuero
Plants of SP 71-6163 variety positive for the virus of the sugarcane yellow leaf syndrome (ScYLV) were sowed under greenhouse conditions to purify the causal agent and to produce specific antibodies. The methodology used for purification was that of Lockhart (1999) modified by CENICAÑA, adding the C...
| Main Authors: | , , , , , , |
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| Format: | Journal Article |
| Language: | Español |
| Published: |
2002
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| Subjects: | |
| Online Access: | https://hdl.handle.net/10568/44374 |
| _version_ | 1855515162870546432 |
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| author | Ángel Sanchez, JC Avellaneda Barbosa, MC Victoria Kafure, JI Betancourt Vasquez, M. Díaz Granados, C. Arroyave, José A. Castaño, M. |
| author_browse | Arroyave, José A. Avellaneda Barbosa, MC Betancourt Vasquez, M. Castaño, M. Díaz Granados, C. Victoria Kafure, JI Ángel Sanchez, JC |
| author_facet | Ángel Sanchez, JC Avellaneda Barbosa, MC Victoria Kafure, JI Betancourt Vasquez, M. Díaz Granados, C. Arroyave, José A. Castaño, M. |
| author_sort | Ángel Sanchez, JC |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | Plants of SP 71-6163 variety positive for the virus of the sugarcane yellow leaf syndrome (ScYLV) were sowed under greenhouse conditions to purify the causal agent and to produce specific antibodies. The methodology used for purification was that of Lockhart (1999) modified by CENICAÑA, adding the Celluclast 1,5 L to 2% enzyme to the extraction buffer. The purified of the obtained virus were evaluated by electronic microscopy in the laboratory of the Virology Unit at CIAT inn order to verify viral particles concentration. Once obtained a high concentration of virus particles, the purified were put on a sucrose gradient with density of 10-40 % to increase the purity of the virus which was determined later in the spectrophotometer. The purified obtained was used as antigen in the production of inmunoglobulin by injecting New Zealand rabbits during four weeks by a concentration of 1mg ScYLV/ml. The antiserum was evaluated by ISEM and TBIA. High concentration of virus particles was found by adding 2 % Celluclast 1,5 L. The absorbance A200/A280 was of 1.49 and the approximate viral concentration of 1.87 mg/ml. The antibody produced will be used for serological tests of ISEM and TBIA in the diagnosis of the ScYLV in the sugarcane area of the Cauca Valley. |
| format | Journal Article |
| id | CGSpace44374 |
| institution | CGIAR Consortium |
| language | Español |
| publishDate | 2002 |
| publishDateRange | 2002 |
| publishDateSort | 2002 |
| record_format | dspace |
| spelling | CGSpace443742025-11-12T05:53:16Z Purificación del virus del síndrome de la hoja amarilla de la caña de azúcar (Sugarcane yellow leaf luteovirus ScYLV) y producción de antisuero Ángel Sanchez, JC Avellaneda Barbosa, MC Victoria Kafure, JI Betancourt Vasquez, M. Díaz Granados, C. Arroyave, José A. Castaño, M. saccharum officinarum sugarcane plant viruses antibodies caña de azúcar virus de las plantas anticuerpos Plants of SP 71-6163 variety positive for the virus of the sugarcane yellow leaf syndrome (ScYLV) were sowed under greenhouse conditions to purify the causal agent and to produce specific antibodies. The methodology used for purification was that of Lockhart (1999) modified by CENICAÑA, adding the Celluclast 1,5 L to 2% enzyme to the extraction buffer. The purified of the obtained virus were evaluated by electronic microscopy in the laboratory of the Virology Unit at CIAT inn order to verify viral particles concentration. Once obtained a high concentration of virus particles, the purified were put on a sucrose gradient with density of 10-40 % to increase the purity of the virus which was determined later in the spectrophotometer. The purified obtained was used as antigen in the production of inmunoglobulin by injecting New Zealand rabbits during four weeks by a concentration of 1mg ScYLV/ml. The antiserum was evaluated by ISEM and TBIA. High concentration of virus particles was found by adding 2 % Celluclast 1,5 L. The absorbance A200/A280 was of 1.49 and the approximate viral concentration of 1.87 mg/ml. The antibody produced will be used for serological tests of ISEM and TBIA in the diagnosis of the ScYLV in the sugarcane area of the Cauca Valley. Plantas de la variedad SP 71-6163 afectadas por el virus del síndrome de la hoja amarilla de la cana de azúcar (ScYLV), se sembraron en condiciones de invernadero para purificar el agente causal y producir anticuerpos específicos. La metodología de purificación utilizada fue la de Lockhart (1999) modificada, agregando la enzima Celluclast 1,5 L al 2% a la solución de extracción. Las preparaciones parcialmente purificadas fueron evaluados por microscopia electrónica en el laboratorio de la unidad de virología del CIAT para verificar la concentración de partículas virales. Una vez obtenida una alta concentración el semipurificado se ultracentrifugó en un gradiente de densidad de sucrosa de 10-40% para incrementar la pureza del virus. El purificado obtenido se empleó como antígeno en la producción de immunoglobulinas especificas inyectando conejos de la raza Nueva Zelandia durante cuatro semanas en una concentración de 1mg ScYLV/ml, para luego evaluar el antisuero mediante técnicas de microscopia electrónica y TBIA. Se encontró una alta concentración de partículas virales en semipurificados empleando Celluclast 1,5 L al 2%. La relación de absorbancia A260/A280 fue de 1,49 y la concentración viral aproximada de 1,87 mg/ml. El anticuerpo producido se utilizará en pruebas serológicas de ISEM y tissue-blot para el diagnóstico del ScYLV en la zona azucarera del Valle del Cauca. 2002 2014-10-02T08:33:40Z 2014-10-02T08:33:40Z Journal Article https://hdl.handle.net/10568/44374 es Open Access application/pdf |
| spellingShingle | saccharum officinarum sugarcane plant viruses antibodies caña de azúcar virus de las plantas anticuerpos Ángel Sanchez, JC Avellaneda Barbosa, MC Victoria Kafure, JI Betancourt Vasquez, M. Díaz Granados, C. Arroyave, José A. Castaño, M. Purificación del virus del síndrome de la hoja amarilla de la caña de azúcar (Sugarcane yellow leaf luteovirus ScYLV) y producción de antisuero |
| title | Purificación del virus del síndrome de la hoja amarilla de la caña de azúcar (Sugarcane yellow leaf luteovirus ScYLV) y producción de antisuero |
| title_full | Purificación del virus del síndrome de la hoja amarilla de la caña de azúcar (Sugarcane yellow leaf luteovirus ScYLV) y producción de antisuero |
| title_fullStr | Purificación del virus del síndrome de la hoja amarilla de la caña de azúcar (Sugarcane yellow leaf luteovirus ScYLV) y producción de antisuero |
| title_full_unstemmed | Purificación del virus del síndrome de la hoja amarilla de la caña de azúcar (Sugarcane yellow leaf luteovirus ScYLV) y producción de antisuero |
| title_short | Purificación del virus del síndrome de la hoja amarilla de la caña de azúcar (Sugarcane yellow leaf luteovirus ScYLV) y producción de antisuero |
| title_sort | purificacion del virus del sindrome de la hoja amarilla de la cana de azucar sugarcane yellow leaf luteovirus scylv y produccion de antisuero |
| topic | saccharum officinarum sugarcane plant viruses antibodies caña de azúcar virus de las plantas anticuerpos |
| url | https://hdl.handle.net/10568/44374 |
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