An enzyme-linked immunosorbent assay (ELISA) for detection of Clostridium aldrichii in anaerobiodigesters.
Monoclonal antibodies (Mabs) against Clostridium aldrichii were prepared by in vivo and in vitro immunization with whole cells and produced after fusion as ascites in BALB/c mice. An enzyme-linked immunosorbent assay (ELISA) was used to test for specificity and sensitivity of the Mabs to detect Cl. aldrichii. The lower limit for Cl. aldrichii detection in pure and mixed culture with ELISA was 10(5) cells ml-1. Twenty other species of bacteria, including 12 cellulolytic species, were tested for cross-reactivity with the ELISA, but none was detected. The ELISA was used for detection of Cl. aldrichii over a 16-month period in five mesophilic continuously-stirred tank reactors (CSTR) with wood, glucose, sludge or sorghum as substrates. The population of Cl. aldrichii in the poplar wood anaerobic digester effluent was 10(6)-10(7) cells ml-1 over that time. These numbers were confirmed by anaerobic microbiological methods. Results from the ELISA technique were obtained in 36 h vs 3 weeks for culture methods. It is concluded that the ELISA is a useful, time-saving method for identification, detection and quantification of Cl. aldrichii in axenic, mixed culture, and in complex undefined cultures such as those found in anaerobic digesters.
Brigmon
R L
RL
Department of Environmental Engineering Sciences, University of Florida, Gainesville.
Chynoweth
D P
DP
Yang
J C
JC
Zam
S G
SG
D000693:Anaerobiosis; D000818:Animals; D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000942:Antigens, Bacterial / Q000032:analysis; D001431:Bacteriological Techniques / Q000706:statistics & numerical data; D003013:Clostridium / Q000276:immunology / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000706:statistics & numerical data; D000939:Epitopes / Q000032:analysis; D005069:Evaluation Studies as Topic; D051379:Mice; D008807:Mice, Inbred BALB C; D012680:Sensitivity and Specificity
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D000939:Epitopes