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Clinical and Diagnostic Laboratory Immunology, May 2004, p. 446-451, Vol. 11, No. 3
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.3.446-451.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Use of Monoclonal Antibodies That Recognize p60 for Identification of Listeria monocytogenes

Kang-Y. Yu,1,{dagger} Youngsoon Noh,1,{dagger} Minsub Chung,1,{dagger} Hong-J. Park,1 Namseok Lee,1 Moonyeon Youn,1 Byeong Y. Jung,2 and Byung-S. Youn1*

KOMED Institute for Life Science, Graduate School of Biotechnology, Korea University,1 Bacteriology and Parasitology Division, National Veterinary Research and Quarantine Service, Gyonggido, Korea2

Received 25 August 2003/ Returned for modification 19 November 2003/ Accepted 13 January 2004

Listeria monocytogenes causes major food-borne outbreaks of disease worldwide. Specific identification of this microorganism is of utmost importance to public health and industry. Listeria species are known to secrete a 60-kDa protein collectively termed p60, which is encoded by the iap (invasion-associated protein) gene and secreted in large quantities into the growth media. p60 is a highly immunogenic murein hydrolase that is essential for cell division. Due to these properties, p60 is an ideal diagnostic target for the development of immunological detection systems for L. monocytogenes. We report here two independent lines of monoclonal antibody (MAb): p6007, which specifically recognizes L. monocytogenes p60, and p6017, which reacts with a wide range of Listeria p60 proteins. By combining these antibodies with a polyclonal antibody, we developed efficient sandwich enzyme-linked immunosorbent assay (ELISA) systems which can specifically identify L. monocytogenes or generally detect Listeria species. Since an excess amount of the peptide corresponding to PepA or PepD did not interfere with the ELISA, and direct ELISAs were unable to detect both peptides, we concluded that the epitope presumed to be recognized by p6007 or p6017 could be distinguished from PepA and PepD as described by Bubert et al. (Appl. Environ. Microbiol. 60:3120-3127, 1997). To our best knowledge, this is the first example of an immunological identification system that uses p60-recognizing MAbs.

* Corresponding author. Mailing address: KOMED Institute for Life Science, Graduate School of Biotechnology, Korea University, Rm. 640, 1,5-ka, Anam-dong, Sungbuk-ku, Seoul 136-701, Korea. Phone: 82-2-927-1470. Fax: 82-2-926-1670. E-mail: bsyoun{at}komed.com.

{dagger} K.-Y.Y., Y.N., and M.C. contributed equally to the work.

Clinical and Diagnostic Laboratory Immunology, May 2004, p. 446-451, Vol. 11, No. 3
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.3.446-451.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.





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