Skip to main content
SpringerLink
Log in

Development of an indirect competitive ELISA for detection ofCampylobacter jejuni subsp.jejuni O:23 in foods

  • Published:
Folia Microbiologica Aims and scope Submit manuscript

Abstract

An indirect enzyme immunoassay for rapid detection ofCampylobacter jejuni subsp.jejuni O: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 µg/mL, and 8 µg/mL, respectively. Under such conditions, the detection limit reached 50 CFU/µL, limit of quantification being 480 CFU/µL. 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 ofC. sputorum subsp.sputorum (21.5 %) and with G+ bacteriumMicrococcus 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 toČSN ISO 10272 and commercially available Singlepath® Campylobacter GLISA-Rapid Test.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

ABS:

2,2′-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)

BAN:

bi(4,4′-anisidine) (3,3′-dimethoxybenzidine)

BDA:

1,2-benzenediamine

BSA:

bovine serum albumin

DAB:

3-dimethylaminobenzoic acid

HRP:

horseradish peroxidase

MBH:

3-methyl-2-benzothiazolinone hydrazone

PBS:

phosphate buffered saline

TMB:

3,3′,5,5′-tetramethylbenzidine

References

  • Blaser M.J., Cravens J., Powers B.W., Wang W.-L.L.:Campylobacter enteritidis associated with canine infection.Lancetii, 979–981 (1978).

    Article  Google Scholar 

  • Blaser M.J., Waldman R.J., Barrett T., Erlandson A.L.: Outbreaks ofCampylobacter enteritis in two extended families — evidence for person-to-person transmission.J.Pediatr.98, 254–257 (1981).

    Article  PubMed  CAS  Google Scholar 

  • Birkenhead D., Hawkey P.M., Heritage J., Gascoyne-Binzi D.M., Kite P.: PCR for detection and typing ofCampylobacters.Lett.Appl.Microbiol.17, 235–237 (1993).

    Article  PubMed  CAS  Google Scholar 

  • Bolton F.J., Hutchinson D.N., Coates D.: Blood-free selective medium for isolation ofCampylobacter jejuni from feces.J.Clin.Microbiol.19, 169–171 (1984).

    PubMed  CAS  Google Scholar 

  • Bryan F.L., Doyle M.P.: Health risks and consequences ofSalmonella andCampylobacter jejuni in raw poultry.J.Food Protect.58, 326–344 (1995).

    Google Scholar 

  • ČSN ISO 10272: Microbiology of Food and Animal Stuffs — Horizontal Method for Detection of ThermotolerantCampylobacter (1997).

  • Dedič K., Bockemuhl J., Kühn H., Volkmer K.-J., Weinke U.T.: Campylobacteriosen, pp. 49–65 inBakterielle Zoonosen Bei Tier und Mensch. Ferdinand-Enke-Verlag, Stuttgart 1993.

    Google Scholar 

  • Docherty L., Adams M.R., Patel P., McFadden J.: The magnetic immuno-polymerase chain reaction assay for the detection ofCampylobacter in milk and poultry.Lett.Appl.Microbiol.22, 288–292 (1996).

    Article  PubMed  CAS  Google Scholar 

  • Hochel I., Jeníková G., Dursi C.F., Pazlarová J., Girotti S., Demnerová K.: Application of mouse antibodies to somatic antigen for detection ofSalmonella enteritidis by competitive ELISA.Food Agric.Immunol.13, 115–126 (2001).

    Article  CAS  Google Scholar 

  • Hodinka R.L., Gilligan P.H.: Evaluation of the Campyslide agglutination test for confirmatory identification of selectedCampylobacter species.J.Clin.Microbiol.26, 47–49 (1988).

    PubMed  CAS  Google Scholar 

  • Humprey T.J.: Techniques for the optimum recovery of cold injuredCampylobacter jejuni from milk or water.J.Appl.Bacteriol.61, 125–132 (1986).

    Google Scholar 

  • Humprey T.J., Henley A., Lanning U.D.G.: The colonization of broiler chickens withCampylobacter jejuni; some epidemiological investigations.Epidemiol.Infect.110, 601–607 (1993).

    Google Scholar 

  • Jackson T.M., Marshall N.J., Etkins R.P.: Optimization of immunoradiometric (labelled antibody) assays, pp. 557–575 in W.M. Hunter, J.E.T. Corrie (Eds):Immunoassays for Clinical Chemistry. Churchill Livingstone, Edinburgh 1983.

    Google Scholar 

  • Jackson C.J., Fox A.J., Jones D.M.: A novel polymerase chain reaction assay for the detection and specification of thermophilicCampylobacter spp.J.Appl.Bacteriol.81, 467–473 (1996).

    PubMed  CAS  Google Scholar 

  • Jacob L., Schmitt E., Bruemmer W.: Purification of antibodies by new chromatography techniques.Am.Biotechnol.Lab.12, 44–45 (1994).

    PubMed  CAS  Google Scholar 

  • Karmali M.A., Simor A.E., Roscoe M., Fleming P.C., Smith S.S., Lane J.: Evaluation of blood-free, charcoal-based, selective medium for the isolation ofCampylobacter organisms from feces.J.Clin.Microbiol.23, 456–459 (1986).

    PubMed  CAS  Google Scholar 

  • Karpinsky K.F.: Optimality assessment in the enzyme-linked immunosorbent assay (ELISA).Biometrics46, 381–390 (1990).

    Article  Google Scholar 

  • Lior H., Woodward D.L., Edgar J.A., Laroche L.J., Gill P.: Serotyping ofCampylobacter jejuni by slide agglutination based on heat-labile antigenic factors.J.Clin.Microbiol.15, 761–768 (1982).

    PubMed  CAS  Google Scholar 

  • Lu P., Brooks B.W., Robertson R.H., Nielsen K.H., García M.M.: Characterization of monoclonal antibodies for the rapid detection of food borne campylobacters.Internat.J.Food Microbiol.37, 87–91 (1997).

    Article  Google Scholar 

  • Mentzing L.O.: Waterborne outbreaks ofCampylobacter enteritis in central Sweden.Lancetii, 352–354 (1981).

    Article  Google Scholar 

  • Mills S.D., Congi R.V., Hennessy J.N., Penner J.L.: Evaluation of a simplified procedure for serotypingCampylobacter jejuni andCampylobacter coli which is based on the O antigen.J.Clin.Microbiol.29, 2093–2098 (1991).

    PubMed  CAS  Google Scholar 

  • Ng L.K., Kingombe I.B., Yan W., Taylor D.E., Hiratsuka K., García M.M.: Specific detection and confirmation ofCampylobacter jejuni by DNA hybridization and PCR.Appl.Environ.Microbiol.63, 4558–4563 (1997).

    PubMed  CAS  Google Scholar 

  • Nielsen E.M., Engberg J., Madsen M.: Distribution of serotypes ofCampylobacter jejuni andC. coli from Danish patients, poultry, cattle and swine.FEMS Immunol.Med.Microbiol.19, 47–56 (1997).

    PubMed  CAS  Google Scholar 

  • Park C.E., Stankiewitz Z.K., Lovett J., Hunt J.: Incidence ofCampylobacter jejuni in fresh eviscerated whole market chickens.Can.J.Microbiol.27, 841–842 (1981).

    Article  PubMed  CAS  Google Scholar 

  • Penner J.L., Hennessy J.N.: Passive hemagglutination technique for serotypingCampylobacter fetus subsp.jejuni on the basis of soluble heat-stable antigens.J.Clin.Microbiol.12, 732–737 (1980).

    PubMed  CAS  Google Scholar 

  • Penner J.L., Hennessy J.N., Congi R.V.: Serotyping ofCampylobacter jejuni andCampylobacter coli on the basis of thermostable antigens.Eur.J.Clin.Microbiol.2, 378–383 (1983).

    Article  PubMed  CAS  Google Scholar 

  • Porstmann T., Porstmann B.: Chromogenic substrates for enzyme immunoassay, pp. 57–84 inNonisotopic Immunoassay (T.T. Ngo, Ed.). Plenum Press, New York 1988.

    Google Scholar 

  • Porstmann T., Porstmann B., Wietschke R., von Baehr R., Egger E.: Stabilization of the substrate reaction of horseradish peroxidase witho-phenylenediamine in the immunoassay.J.Clin.Chem.Clin.Biochem.23, 41–44 (1985).

    PubMed  CAS  Google Scholar 

  • Prusak-Sochaczewski E., Luong J.H.T.: An improved ELISA method for detection ofSalmonella typhimurium.J.Appl.Bacteriol.66, 127–135 (1989).

    PubMed  CAS  Google Scholar 

  • Rice B.E., Rollins D.M., Mallinson E.T., Carr L., Joseph S.W.:Campylobacter jejuni in broiler chickens: colonization and humoral immunity following oral vaccination and experimental infection.Vaccine15, 1922–1932 (1997).

    Article  PubMed  CAS  Google Scholar 

  • Robinson D.A., Jones D.M.: Milk-borneCampylobacter infection.Brit.Med.J.282, 1374–1376 (1981).

    Article  CAS  Google Scholar 

  • Skirrow M.B.:Campylobacter enteritis: a “new” disease.Brit.Med.J.2, 9–11 (1977).

    PubMed  CAS  Google Scholar 

  • Skirrow M.B., Turnbull G.L., Walker R.E., Young S.E.J.:Campylobacter jejuni enteritis transmitted from cat to man.Lanceti, 1188 (1980).

    Article  Google Scholar 

  • Steinhauserová I., Fojtíková K., Klimeš J.: The incidence and PCR detection ofCampylobacter upsaliensis in dogs and cats.Lett.Appl.Microbiol.31, 209–212 (2000).

    Article  PubMed  Google Scholar 

  • Studahl A., Andersson Y.: Risk factors for indigenousCampylobacter infections: a Swedish case-control study.Epidemiol.Infectol.125, 269–275 (2000).

    Article  CAS  Google Scholar 

  • Svedhem A., Norkrans G.:Campylobacter jejuni enteritis transmitted from cat to man.Lanceti, 713–714 (1980).

    Google Scholar 

  • Tijjsen P., Su D.-M., Kurstak E.: Rapid and sensitive heterologous enzyme immunoassays for densonucleosis virus (Parvoviridae).Arch.Virol.74, 277–291 (1982).

    Article  Google Scholar 

  • Wedderkopp A., Gradel K.O., Jørgensen J.C., Madsen M.: Pre-harvest surveillance ofCampylobacter andSalmonella in Danish broiler flocks: 2-year study.Internat.J.Food Microbiol.68, 53–59 (2001).

    Article  CAS  Google Scholar 

  • Wegmüller B., Lüthy J., Candrian U.: Direct polymerase chain reaction detection ofCampylobacter jejuni andCampylobacter coli in raw milk and dairy products.Appl.Environ.Microbiol.59, 2961–2965 (1993).

    Google Scholar 

  • Willis W.L., Murray C.:Campylobacter jejuni seasonal recovery observations of retail market broilers.Poultry Sci.76, 314–317 (1997).

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biochemistry and Microbiology, Institute of Chemical Technology, 166 28, Prague 6, Czechia

    I. Hochel, D. Viochna & M. Musil

  2. Department of Anthropology and Human Genetics, Faculty of Science, Charles University, Prague 2, Czechia

    J. Škvor

Authors
  1. I. Hochel
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. D. Viochna
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. Škvor
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M. Musil
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

The project no. 525/02/0287 (Development of Screening ELISA and PCR Methods for Detection of Campylobacteria in Foods) was supported byGrant Agency of the Czech Republic.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hochel, I., Viochna, D., Škvor, J. et al. Development of an indirect competitive ELISA for detection ofCampylobacter jejuni subsp.jejuni O:23 in foods. Folia Microbiol 49, 579–586 (2004). https://doi.org/10.1007/BF02931537

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02931537

Keywords

Search

Navigation