PRODUCTION, CHARACTERIZATION AND APPLICATION OF MONOCLONAL ANTIBODIES AGAINST XANTHOMONAS CAMPESTRIS PV. MANGIFERAE-INDICAE, X. C. PV. VESICATORIA AND RALSTONIA SOLANACEARUM
K TSUCHIHA, C D'URSEL, F IFTIKHAR, M HORITA and Y NOZU
National Institute of Agrobiological Resources (NIAR), 2-1-1, Kannondai, Tsukuba,305-0856, Japan
Background and objectives
Materials and methods
Results and conclusions
Three representative MABs, 7AH10, 5HB3 and 4AD2, were selected from a total of 90 hybridomas raised against XCV strains UPB141, MAFF 301294 and 2625. All of them were IgM. The dilution limit of 7AH10 and 5HB3 was 125,000, and that of 4AD2 was 24,000. For all the MABs the detection limit was 10 times lower when a competitive ELISA format was used, and then comprised between 104 and 103 c.f.u./ml. With the 32 XCV strains isolated from Japan, Guadeloupe, Maroc, Senegal, Reunion or France, 30 strains (94%) reacted with at least two of three MABs. Particularly, the 17 from Japan were all recognized, and strains of tomato and pepper reacted specificaly with 4AD2 and 7AH10 , respectively. The MAB 7AH10, obtained against the UPB141, an Amy- strain reacted with all the Amy- (n=20) and 15% (n=19) of the Amy+ strains. And against the 2625 strain, an Amy- strain, the MAB 5HB3 recognized 94% of all the XCV (n=39). For all the MABs, cross-reactions with other pathovars or species were less than 4% (n=67). In immuno-blotting, the three MABs reacted with typical LPS bands between 41 and 120 kDa from EDTA extract and water phase after phenol extraction. Thirteen MABs were established from an immunization of live cells of an RS strain MAFF 301520 (race 1, biovar 4) . They were all IgM. They were divided into groups based on reactivity to the strains from tobacco, potato, ginger and eggplant.