A monoclonal antibody-based ELISA was developed to detect alpha-toxin present in Clostridium perfringens bacterial cell lysates and cell-free culture supernatants, Monoclonal antibodies against C. perfringens alpha-toxin were produced in hybridoma tissue culture supernatants and in BALB/c mice ascites fluid, The monoclonal antibodies obtained from hybridoma culture supernatant and ascites fluid showed identical antigen specificity, but the latter showed a higher titer, with a 50% endpoint at 1/4,000. The monoclonal antibodies were specific for phospholipase C produced by C. perfringens, but not by Bacillus cereus. The lower limit of phospholipase C detection was 16 ng/ml. The dose-dependent relationship between absorption at 490 nm and concentration of phospholipase C diluted in HEPES (N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid) or Trypticase glucose yeast broth fit a four-parameter and a quadratic model, respectively. The monoclonal antibody-based ELISA developed is a rapid, sensitive and specific detection method and can be used for quantitative characterization of C. perfringens alpha-toxin.
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Research Article| June 01 1996
Monoclonal Antibody-Based ELISA for Detection of Clostridium perfringens Alpha-Toxin
ZONGLIN L. LIU ;
HANS P. BLASCHEK
HANS P. BLASCHEK *
Department of Food Science & Human Nutrition, University of Illinois, 580 Bevier Hall, 905 S. Goodwin Ave., Urbana, Illinois 61801, USA
* Author for correspondence. Tel: 217-333-8224; Fax: 217-244-2517; Email: email@example.com.
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ZONGLIN L. LIU, HANS P. BLASCHEK; Monoclonal Antibody-Based ELISA for Detection of Clostridium perfringens Alpha-Toxin. J Food Prot 1 June 1996; 59 (6): 621–625. doi: https://doi.org/10.4315/0362-028X-59.6.621
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