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Novel and Highly Specific Monoclonal Antibody to Acidovorax citrulli and Development of ELISA-Based Detection in Cucurbit Leaves and Seed
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Metadata
Document Title
Novel and Highly Specific Monoclonal Antibody to Acidovorax citrulli and Development of ELISA-Based Detection in Cucurbit Leaves and Seed
Author
Himananto O, Thummabenjapone P, Luxananil P, Kumpoosiri M, Hongprayoon R, Kositratana W, Gajanandana O
Name from Authors Collection
Scopus Author ID
23110409300
Affiliations
National Science & Technology Development Agency - Thailand; National Center Genetic Engineering & Biotechnology (BIOTEC); Kasetsart University; Kasetsart University; Khon Kaen University
Type
Article
Source Title
PLANT DISEASE
ISSN
0191-2917
Year
2011
Volume
95
Issue
9
Page
1172-1178
Open Access
hybrid
Publisher
AMER PHYTOPATHOLOGICAL SOC
DOI
10.1094/PDIS-12-10-0889
Format
Abstract
Himananto, O., Thummabenjapone, P., Luxananil, P., Kumpoosiri, M., Hongprayoon, R., Kositratana, W., and Gajanandana, O. 2011. Novel and highly specific monoclonal antibody to Acidovorax citrulli and development of ELISA-based detection in cucurbit leaves and seed. Plant Dis. 95:1172-1178. A novel monoclonal antibody (MAb) specific to the seedborne bacterium Acidovorax citrulli was produced. MAb 11E5 reacted specifically with 19 strains of A. citrulli but not with three closely related bacteria in the family Comamonadaceae (i.e., A. facilis. Comamonas acidovorans, and C. testosteroni) and another seven phytopathogenic bacteria. Moreover, this MAb detected a strain of A. citrulli that was not detected by a commercial enzyme-linked immunosorbent assay (ELISA)-based kit and a commercial immunochromatographic strip test. In Western blot analysis, MAb 11E5 reacted with an A. citrulli protein of a molecular mass >170 kDa. MAb 11E5 was employed to develop two sandwich ELISA systems: MAb captured-sandwich ELISA (MC-sELISA) and polyclonal antibody captured-sandwich ELISA (PC-sELISA). MC-sELISA was 10 times more sensitive than PC-sELISA for detection of A. citrulli in cucurbit leaf and seed extracts. The detection limit of the MC-sELISA was 5 x 10(4) CFU/ml. Detection of A. citrulli in naturally infected cucurbit leaves, fruit, and seed was also feasible using MC-sELISA. The newly established MC-sELISA provides another alternative for specific detection of A. citrulli in cucurbits and can be applied for routine field inspection.
Funding Sponsor
National Center for Genetic Engineering and Biotechnology (BIOTEC, Thailand); Center for Agricultural Biotechnology, Kasetsart University; Kamphaeng Sean Campus/Center of Excellence on Agricultural Biotechnology: (AG-BIO/PERDO-CHE) Thailand
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WOS