Development of an indirect competitive ELISA for detection of Campylobacter jejuni subsp.jejuni O:23 in foods
Language English Country United States Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
15702549
DOI
10.1007/bf02931537
Knihovny.cz E-resources
- MeSH
- Campylobacter jejuni immunology isolation & purification pathogenicity MeSH
- Enzyme-Linked Immunosorbent Assay methods statistics & numerical data MeSH
- Enteritis microbiology MeSH
- Campylobacter Infections microbiology MeSH
- Rabbits MeSH
- Chickens MeSH
- Humans MeSH
- Food Microbiology * MeSH
- Antibodies, Bacterial MeSH
- Sensitivity and Specificity MeSH
- Cross Reactions MeSH
- Animals MeSH
- Check Tag
- Rabbits MeSH
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Czech Republic MeSH
- Names of Substances
- Antibodies, Bacterial MeSH
An indirect enzyme immunoassay for rapid detection of Campylobacter jejuni subsp. jejuni 0:23 has been developed. Optimum concentrations of immobilized cells, polyclonal chicken IgY, and rabbit anti-IgY antibody-horseradish peroxidase conjugate were 3.1 CFU/nL, 10 microg/mL, and 8 microg/mL, respectively. Under such conditions, the detection limit reached 50 CFU/microL, limit of quantification being 480 CFU/microL. By testing 5 chromogens, viz. 1,2-benzenediamine, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid), 3,3',5,5'-tetramethylbenzidine, bi(4,4'-anisidine) and 3-methyl-2-benzothiazolinone hydrazone, in horseradish peroxidase substrate, 1,2-benzenediamine or 3,3',5,5'-tetramethylbenzidine as H-donors in the enzyme substrate provided the highest ELISA sensitivity. The applied polyclonal antibody was specific for homogeneous antigen. The cross-reactions were observed only with one strain of C. sputorum subsp. sputorum (21.5 %) and with G+ bacterium Micrococcus luteus (6.1 %). Preliminary tests have been performed with a limited number of artificially contaminated food samples. No matrix effects on the ELISA sensitivity were observed. The results (by means of ELISA) were comparable with those given by both a standard cultivation method performed according to CSN ISO 10272 and commercially available Singlepath Campylobacter GLISA-Rapid Test.
Department of Biochemistry and Microbiology Institute of Chemical Technology 166 28 Prague 6 Czechia
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