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Copyright ©The Author(s) 2004. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Sep 15, 2004; 10(18): 2747-2749
Published online Sep 15, 2004. doi: 10.3748/wjg.v10.i18.2747
Development of an ELISA kit using monoclonal antibody to Clostridium difficile toxin A
Si-Wu Fu, Ya-Li Zhang, Dian-Yuan Zhou
Si-Wu Fu, Ya-Li Zhang, Dian-Yuan Zhou, PLA Institute for Digestive Medicine, Nanfang Hospital, First Military Medical University, Guangzhou 510515, Guangdong province, China
Author contributions: All authors contributed equally to the work.
Correspondence to: Dr. Si-Wu Fu, MD. PLA Institute for Digestive Medicine, Nanfang Hospital, First Military Medical University, Guangzhou 510515, Guangdong Province, China. fusiwu2001@sina.com
Telephone: +86-20-61641544
Received: December 28, 2003
Revised: January 5, 2004
Accepted: January 12, 2004
Published online: September 15, 2004
Abstract

AIM: To establish an ELISA kit using monoclonal antibodies against Clostridium difficile (C. difficile) toxin A.

METHODS: An indirect sandwich ELISA was described using the purified rabbit monospecific antiserum as capturing antibody. After the polystyrene microtitre plates with 96 flat-bottomed wells were coated with rabbit antiserum, the wells were blocked with 100 g/L BSA in PBS-T. C. difficile toxin A or culture filtrates were added to each well and then monoclonal antibodies IgG-horseradish peroxidase conjugate was added as detecting antibody, tetramethylbenzidine was used as substrate and A450 of the stopped reacting product was recorded in an automated plate reader.

RESULTS: The tested specimens included culture filtrates of 2 strains of toxigenic C. difficile, 2 strains of non-toxigenic C. difficile, 26 strains of E. coli, 2 strains of S. dysenteriae, 1 strain of Bif. infantis, 5 strains of V. cholera, 2 strains of S. typhi, 7 strains of C. botulinum, 1 strain of toxigenic C. sordllii, and 1 strain of C. butyricum. A total of 47 strains of culture filtrates were all negative except for 2 strains of toxigenic C. difficile. The detective limitation of toxin A was 0.1 ng/mL.

CONCLUSION: An ELISA kit with high specificity and excellent sensitivity for the rapid detection of C. difficile toxin A was established. It will be a useful tool for diagnostic test of C. difficile toxin A.

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