An ELISA Test for Bacterial Ring Rot of Potato with a New Monoclonal Antibody

Detection of Clavibacter michiganensis subsp. sepedonicus by AmpliDet RNA, a new technology based on real time monitoring of NASBA amplicons with a molecular beacon

“…Direct plating generally results in an overgrowth of Cms by more rapidly-growing bacteria. The serological tests include immunofluorescence cell staining (De Boer and Copeman 1980), immunofluorescence colony staining (Roozen and Van Vuurde 1991) and ELISA (De Boer et al 1988). These tests are fairly sensitive, with a detection level of about 10 000 cells ml )1 , but they may occasionally give falsepositive results due to cross-reactions.…”

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“…118 Vol. 38, Plant Protection Science -2002 bodies because they are produced to single determinants (DE BOER et al 1988). They can replace polyclonal antisera in any one of the serological diagnostic procedures.…”

“…In contrast to data obtained with monoclonal antibodies, those gained with polyclonal antisera were often conflicting and un-convicting due to the occurrence of false positive cross-reactions. The use of monoclonal antibodies, in comparison with polyclonal antibodies, is also complicated by the appearence of cross-reactions with various pathogenic and saprophytic soil bacteria found in potato extracts.Research institutes and laboratories of plant health services dealing with the diagnosis of the ring rot pathogen buy antibodies for immunochemical diagnosis of Cms available on the market and/or prepare their own antibodies (DE BOER et al 1988;WESTRA et al 1994). The use of more than one antibody for detection of the target bacterium in plant samples excludes even more potential crossreactions.…”

“…The pellet was dissolved in 1% formaldehyde solution, incubated overnight at 24°C, and then the suspension was centrifuged. The concentration of antigen was adjusted for immunization and used for monoclonal antibody evaluation in the range 10 7 -10 3 cfu/ml.For purification of extracellular polysaccharides, the method previously reported by DE BOER et al (1988) was used. For immunisation and evaluation of monoclonal antibody, the concentration of the extracellular polysaccharides was adjusted in a range 0.1-2.0 mg/ml by the phenol-sulphuric acid assay using glucose as the standard (ANONYMOUS 1983).…”

“…Better antibodies and prior treatment of antigen did not result in a stronger ELISA reaction, and a more reliable determination and detection of Cms by polyclonal antibodies in ELISA did not seem feasible (KOKOŠKOVÁ et al 2000). The preparation of monoclonal antibodies rather than polyclonal antibodies for plant pathogenic bacteria of the genus Clavibacter has been definitely preferred in the last 10 years (ERWING 1990).Previous studies by WIECZOREK (1984) andDE BOER et al (1988) with polyclonal antibodies showed that Cms possesses a soluble antigen of diagnostic value that may perform better than cell wall antigens in some serological tests. They produced a monoclonal antibody to that soluble antigen of Cms and evaluated its usefulness in several serological procedures.…”