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The use of monoclonal antibodies for the identification and quantification of potato cyst nematodes 1
Author(s) -
EVANS K.,
CURTIS R. H.,
ROBINSON M. P.,
YEUNG M.
Publication year - 1995
Publication title -
eppo bulletin
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.327
H-Index - 36
eISSN - 1365-2338
pISSN - 0250-8052
DOI - 10.1111/j.1365-2338.1995.tb01478.x
Subject(s) - monoclonal antibody , globodera rostochiensis , antigenicity , antigen , biology , cyst , heterodera , nematode , globodera pallida , immunoassay , potato cyst nematode , sonication , urea , antibody , chromatography , veterinary medicine , chemistry , biochemistry , immunology , pathology , ecology , medicine , solanaceae , gene
Experiments were conducted on methods of preparation of samples of cysts, eggs and larvae of Globodera rostochiensis and G. pallida in order to improve the efficiency of immunoassay as a tool for identification and quantification of nematodes in soil. Monoclonal antibodies, which specifically recognize each of the two species of potato cyst nematode and do not cross‐react with other species of soil nematodes, were used in ELISA tests. It was shown that free larvae react less well to monoclonal antibodies than cysts and eggs and that a large proportion of the relevant antigens comes from the vitelline fluid of eggs. Dead eggs and non‐viable components of the cyst (e.g. cyst wall, egg shell) have little or no antigenicity. Of 13 different chemical extractants tested for release of antigens from cysts, CTAB and especially 1% urea were shown to be more effective than the standard extractant PBS. Sonication of cysts was the best of a number of chemical, physical and antigen release procedures. There was a good quantitative relationship between optical density readings, in indirect and DAS‐ELISA, and numbers of nematodes extracted from soil, but further work is needed in order for the test to work directly on soil samples.