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Applied and Environmental Microbiology, July 2002, p. 3634-3638, Vol. 68, No. 7
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.7.3634-3638.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Enrichment Double-Antibody Sandwich Indirect Enzyme-Linked Immunosorbent Assay That Uses a Specific Monoclonal Antibody for Sensitive Detection of Ralstonia solanacearum in Asymptomatic Potato Tubers

Paola Caruso,1 María Teresa Gorris,1 Mariano Cambra,1 José Luis Palomo,2 Jesús Collar,3 and María M. López1*

Instituto Valenciano de Investigaciones Agrarias, Apartado Oficial, 46113 Moncada, Valencia,1 Centro Regional de Diagnóstico, Apartado 61, 37080 Salamanca,2 Laboratorio Agrario e Fitopatolóxico de Galicia, 15318 Mabegondo, La Coruña, Spain3

Received 15 October 2001/ Accepted 24 March 2002

Sensitive and specific routine detection of Ralstonia solanacearum in symptomless potato tubers was achieved by efficient enrichment followed by a reliable double-antibody sandwich indirect enzyme-linked immunosorbent assay based on the specific monoclonal antibody 8B-IVIA. This monoclonal antibody reacted with 168 typical R. solanacearum strains and did not recognize 174 other pathogenic or unidentified bacteria isolated from potato. The optimized protocol included an initial enrichment step consisting of shaking the samples in modified Wilbrink broth for 72 h at 29°C. This step enabled specific detection by the enzyme-linked immunosorbent assay of 1 to 10 CFU of R. solanacearum per ml of initial potato extract. Analysis of 233 commercial potato lots by this method provided results that coincided with the results of conventional methods.

* Corresponding author. Mailing address: Departamento de Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias, Apartado Oficial, Carretera Moncada-Náquera Km 4.5, 46113 Moncada, Valencia, Spain. Phone: 34-96-1391000. Fax: 34-96-1390240. E-mail: mlopez{at}ivia.es.

Applied and Environmental Microbiology, July 2002, p. 3634-3638, Vol. 68, No. 7
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.7.3634-3638.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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  • Alvarez, B., Lopez, M. M., Biosca, E. G. (2007). Influence of Native Microbiota on Survival of Ralstonia solanacearum Phylotype II in River Water Microcosms. Appl. Environ. Microbiol. 73: 7210-7217 [Abstract] [Full Text]  
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  • Caruso, P., Palomo, J. L., Bertolini, E., Alvarez, B., Lopez, M. M., Biosca, E. G. (2005). Seasonal Variation of Ralstonia solanacearum Biovar 2 Populations in a Spanish River: Recovery of Stressed Cells at Low Temperatures. Appl. Environ. Microbiol. 71: 140-148 [Abstract] [Full Text]  


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