MONOCLONAL-ANTIBODIES FOR USE IN DETECTION OF BACILLUS AND CLOSTRIDIUM SPORES

Five monoclonal antibodies against bacterial spores of Bacillus cereus T and Clostridium sporogenes PA3679 were developed. Two antibodies (B 48 and B183) mere selected for their reactivity with B. cereus T spore s, two (C33 and C225) were selected for their reactivity with C. sporo genes spores, and one (D89) was selected for its reactivity with both B. cereus and C. sporogenes spores. The isotypes of the antibodies wer e determined to be immunoglobulin G2a (IgG2a) (B48), IgG1 (B183), and IgM (C33, C225, and D89). The antibodies reacted with spores of B. cer eus T, Bacillus subtilis subsp, globigii, Bacillus megaterium, Bacillu s stearothermophilus, C. sporogenes, Clostridium perfringens, and Desu lfotomaculum nigrificans. Antibody D89 also reacted with vegetative ce lls of B. cereus and C. sporogenes. Analysis of B. cereus spore extrac ts showed that two of the antigens with which the anti-Bacillus antibo dies reacted had molecular masses of 76 kDa and approximately 250 kDa, Immunocytochemical localization indicated that antigens with which B4 8, B183, and D89 react are on the exosporium of the B, cereus T spore, Antibody D89 reacted with the exosporium and outer cortex of C. sporo genes spores in immunocytochemical localization studies but did not re act with extracts of C. sporogenes or B, cereus spores in Western blot ting. Some C. sporogenes antigens were not stable during long-term sto rage at -20 degrees C. Antibodies B48, B183, and D89 should prove to b e useful tools for developing immunological methods for the detection of bacterial spores.