Detection and Diagnosis of Xylella fastidiosa by Specific Monoclonal Antibodies
Monoclonal antibodies (MAb) specific to Xylella fastidiosa were obtained through hybridoma technology using heat-treated somatic O antigens from LMG 17159strain. Ten stable hybrydoma clones secreting MAb were selected and their isotype was determined. The MAbs 2G1/PPD, IgG1 showed specificity for X....
| Autores principales: | , , , , , |
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| Formato: | publishedVersion |
| Lenguaje: | Inglés |
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MDPI
2021
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| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.11939/6967 https://www.mdpi.com/2073-4395/11/1/48 |
| _version_ | 1855032522754228224 |
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| author | Gorris, María T. Sanz, Antonio Penyalver, Javier López, María M. Colomer, Mario Marco-Noales, Ester |
| author_browse | Colomer, Mario Gorris, María T. López, María M. Marco-Noales, Ester Penyalver, Javier Sanz, Antonio |
| author_facet | Gorris, María T. Sanz, Antonio Penyalver, Javier López, María M. Colomer, Mario Marco-Noales, Ester |
| author_sort | Gorris, María T. |
| collection | ReDivia |
| description | Monoclonal antibodies (MAb) specific to Xylella fastidiosa were obtained through hybridoma technology using heat-treated somatic O antigens from LMG 17159strain. Ten stable hybrydoma clones secreting MAb were selected and their isotype was determined. The MAbs 2G1/PPD, IgG1 showed specificity for X. fastidiosa, detecting all the analyzed strains representing different subspecies, STs and hosts. Polyclonal antibodies (PAb) against X. fastidiosa were also produced and antiserum 17159-O/IVIA was selected for the highest titre and its excellent detection capability. MAb 2G1/PPD was tested against strain IVIA 5235 in PBS and in spiked raw extract samples from almond, olive, citrus, and other hosts and its sensitivity by DAS-ELISA was 104 CFU mL−1. The MAb also reacted with high affinity and avidity against X. fastidiosa by DASI-ELISA and Tissue print-ELISA. The diagnostic parameters of DAS-ELISA based on MAb were calculated and compared with the gold standard real-time PCR. The diagnostic specificity of MAb2G1/PPD was 100%, the diagnostic sensitivity was 88.5% compared to Harper’s real-time PCR and 89.9% compared to Francis’ real-time PCR. The agreement between the techniques was almost perfect according to the estimated Cohen’s kappa-index, even in symptomless almond trees. The developed immunological techniques represent sustainable and low-cost analysis tools, based on specific, homogeneous, and well-characterized MAbs, which can be obtained in unlimited quantities in a reproducible way and constitute a guarantee for the standardization of commercial kits. They are a valuable option within a polyphasic strategy for the detection of X. fastidiosa. |
| format | publishedVersion |
| id | ReDivia6967 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2021 |
| publishDateRange | 2021 |
| publishDateSort | 2021 |
| publisher | MDPI |
| publisherStr | MDPI |
| record_format | dspace |
| spelling | ReDivia69672025-04-25T14:48:01Z Detection and Diagnosis of Xylella fastidiosa by Specific Monoclonal Antibodies Gorris, María T. Sanz, Antonio Penyalver, Javier López, María M. Colomer, Mario Marco-Noales, Ester DAS-ELISA DASI-ELISA Tissue print-ELISA Diagnostic parameters Cohen’s kappa index Sustainable diagnosis H20 Plant diseases ELISA Monoclonal antibodies Monoclonal antibodies (MAb) specific to Xylella fastidiosa were obtained through hybridoma technology using heat-treated somatic O antigens from LMG 17159strain. Ten stable hybrydoma clones secreting MAb were selected and their isotype was determined. The MAbs 2G1/PPD, IgG1 showed specificity for X. fastidiosa, detecting all the analyzed strains representing different subspecies, STs and hosts. Polyclonal antibodies (PAb) against X. fastidiosa were also produced and antiserum 17159-O/IVIA was selected for the highest titre and its excellent detection capability. MAb 2G1/PPD was tested against strain IVIA 5235 in PBS and in spiked raw extract samples from almond, olive, citrus, and other hosts and its sensitivity by DAS-ELISA was 104 CFU mL−1. The MAb also reacted with high affinity and avidity against X. fastidiosa by DASI-ELISA and Tissue print-ELISA. The diagnostic parameters of DAS-ELISA based on MAb were calculated and compared with the gold standard real-time PCR. The diagnostic specificity of MAb2G1/PPD was 100%, the diagnostic sensitivity was 88.5% compared to Harper’s real-time PCR and 89.9% compared to Francis’ real-time PCR. The agreement between the techniques was almost perfect according to the estimated Cohen’s kappa-index, even in symptomless almond trees. The developed immunological techniques represent sustainable and low-cost analysis tools, based on specific, homogeneous, and well-characterized MAbs, which can be obtained in unlimited quantities in a reproducible way and constitute a guarantee for the standardization of commercial kits. They are a valuable option within a polyphasic strategy for the detection of X. fastidiosa. 2021-01-14T08:58:56Z 2021-01-14T08:58:56Z 2021 publishedVersion Gorris, M. T., Sanz, A., Peñalver, J., López, M. M., Colomer, M., & Marco-Noales, E. (2021). Detection and Diagnosis of Xylella fastidiosa by Specific Monoclonal Antibodies. Agronomy, 11(1), 48 http://hdl.handle.net/20.500.11939/6967 10.3390/agronomy11010048 https://www.mdpi.com/2073-4395/11/1/48 en Atribución-NoComercial-SinDerivadas 3.0 España http://creativecommons.org/licenses/by-nc-nd/3.0/es/ openAccess MDPI electronico |
| spellingShingle | DAS-ELISA DASI-ELISA Tissue print-ELISA Diagnostic parameters Cohen’s kappa index Sustainable diagnosis H20 Plant diseases ELISA Monoclonal antibodies Gorris, María T. Sanz, Antonio Penyalver, Javier López, María M. Colomer, Mario Marco-Noales, Ester Detection and Diagnosis of Xylella fastidiosa by Specific Monoclonal Antibodies |
| title | Detection and Diagnosis of Xylella fastidiosa by Specific Monoclonal Antibodies |
| title_full | Detection and Diagnosis of Xylella fastidiosa by Specific Monoclonal Antibodies |
| title_fullStr | Detection and Diagnosis of Xylella fastidiosa by Specific Monoclonal Antibodies |
| title_full_unstemmed | Detection and Diagnosis of Xylella fastidiosa by Specific Monoclonal Antibodies |
| title_short | Detection and Diagnosis of Xylella fastidiosa by Specific Monoclonal Antibodies |
| title_sort | detection and diagnosis of xylella fastidiosa by specific monoclonal antibodies |
| topic | DAS-ELISA DASI-ELISA Tissue print-ELISA Diagnostic parameters Cohen’s kappa index Sustainable diagnosis H20 Plant diseases ELISA Monoclonal antibodies |
| url | http://hdl.handle.net/20.500.11939/6967 https://www.mdpi.com/2073-4395/11/1/48 |
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