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Serological Detection of Citrus tristeza virus with Antibodies Developed to the Recombinant Coat Protein

January 2009 , Volume 93 , Number  1
Pages  11 - 16

M. M. Iracheta-Cárdenas, Instituto de Biotecnología, Facultad de Ciencias Biológicas, UANL, Pedro de Alba s/n, Cd. Universitaria, San Nicolás de los Garza, Nuevo León 66450 México; P. Metheney, Central California Tristeza Eradication Agency, and M. L. Polek, California Department of Food and Agriculture, Tulare 93274; K. L. Manjunath and R. F. Lee, United States Department of Agriculture--Agricultural Research Service, National Clonal Germplasm Repository for Citrus and Dates, Riverside, CA, 92507; and M. A. Rocha-Peña, INIFAP/UANL, Unidad de Investigación en Biología Celular y Molecular, Apartado Postal 128-F, Cd. Universitaria, San Nicolás de los Garza, Nuevo León 66450 México

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Accepted for publication 30 August 2008.

Antibodies specific for the recombinant coat protein (rCP) of the p25 gene of Citrus tristeza virus (CTV) were developed in goats and rabbits and further evaluated as a complete kit for the detection of the virus using healthy and CTV-infected tissue. The combination of goat T1 used as primary (coating) and rabbit C3 as intermediate (detecting) rCP antibodies reacted efficiently, with optical density at 405 nm (OD405) values between 0.250 and 2.000 with samples from an international collection of diverse CTV isolates. The CTV isolates tested cause a broad spectrum of disease syndromes in different citrus hosts. The OD405 values for healthy tissue were less than 0.100. Likewise, the combination of goat T1 and rabbit C3 rCP antibodies gave consistent results for CTV-positive and -negative sample discrimination when directly compared with the Central California Tristeza Eradication Agency (CCTEA) antibodies used for large-scale CTV detection and a commercially available CTV serological detection kit. The combination of goat T1 and rabbit C3 rCP antibodies showed its suitability for large-scale indexing with samples collected in commercial groves as part of the CCTEA's regular monitoring program. The evaluation included 41,195 samples from 301 commercial groves from districts 1, 2, and 3. In total, 26 trees (0.063%) were found to be CTV positive using the T1/C3 rCP antibody combination. Results of this research provide evidence that rCP antibodies can be efficiently used for both capturing and detecting CTV antigens in double-antibody sandwich indirect enzyme-linked immunosorbent assay.

© 2009 The American Phytopathological Society