J Vet Sci.
2005 Mar;6(1):41-46.
Development of a sandwich ELISA for the detection of Listeria spp. using specific flagella antibodies
- Affiliations
-
- 1National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry, Anyang 430-824, Korea.
- 2Department of Veterinary Medicine, Cheju National University, Jeju 690-756, Korea. yklim@cheju.ac.kr
- 3Faculty of Animal Science and Veterinary Medicine, Thainguyen University of Agriculture and Forestry, Thainguyen, Vietnam.
- 4Department of Veterinary Medicine, Seoul National University, Seoul 151-742, Korea.
- 5Department of Biology, College of Natural Science, Kyonggi University, Suwon 442-760, Korea.
Abstract
- Five monoclonal antibodies (MAbs) and chicken immunoglobulin (IgY) were developed by immunizing with flagella purified from Listeria monocytogenes 4b and the five MAbs have been confirmed to be specific against three different epitopes of flagellin. The antibodies showed specific reaction to Listeria genus and no cross-reactivity with other bacteria tested in this experiment including E.coli O157:H7 and Salmonella enteritidis. Sandwich enzyme-linked immunosorbent assays (ELISA) using the MAbs and IgY were developed to detect Listeria species and the sensitivity and specificity of the developed ELISA have been analyzed. The detection limit of ELISA using MAb 2B1 and HRP labeled IgY was 1 x105cells/0.1 ml at 22degrees C and 1x106 cells/0.1 ml at 30degrees C. ELISA using the pair of MAbs (MAbs 2B1 and HRP labeled MAbs 7A3) detected up to 104cells/0.1 ml at 22degrees C and 30degrees C. Detection limit of sandwich ELISA using IgY was 10 times lower than MAb pair. Using the developed ELISA, we could detect several Listeria contaminated in food samples after 48 h-culturing. In conclusion, both MAbs and IgY have been proved to be highly specific to detect Listeria flagella and the developed sandwich ELISA using these antibodies would be useful tool for screening Listeria spp. in food.
Keyword
MeSH Terms
-
Animals
Antibodies, Bacterial/*chemistry
Antibodies, Monoclonal
Antibody Specificity
Antigens, Bacterial/analysis
Enzyme-Linked Immunosorbent Assay/*methods
Flagella/*genetics
Food Microbiology
Immunoglobulins/analysis
Listeria/*classification/immunology/*isolation&purification
Meat/microbiology
Milk/microbiology
Sensitivity and Specificity
Swine
- Full Text Links
-
- Actions
-
Cited
- CITED
-
- Close
- Share
-
- Similar articles
-
- Detection of Infectious Bursal Disease Virus by Double Antibody Sandwich ELISA
- Development of Enzyme Linked Immunosorbent Assay for Erythropoietin
- Production of monoclonal antibodies to lipoarabinomannan-B and use in the detection of mycobacterial antigens in sputum
- Development of a Quantitative Sandwich Enzyme-Linked Immunosorbent Assay for Detecting the MPT64 Antigen of Mycobacterium tuberculosis
- Western blot analysis of stray cat sera against Toxoplasma gondii and the diagnostic availability of monoclonal antibodies in sandwich-ELISA
