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Electrical protein array chips for the detection of staphylococcal virulence factors

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Abstract

A new approach for the detection of virulence factors of Staphylococcus aureus and Staphylococcus epidermidis using an electrical protein array chip technology is presented. The procedure is based on an enzyme-linked sandwich immunoassay, which includes recognition and binding of virulence factors by specific capture and detection antibodies. Detection of antibody-bound virulence factors is achieved by measuring the electrical current generated by redox recycling of an enzymatically released substance. The current (measured in nanoampere) corresponds to the amount of the target molecule in the analyzed sample. The electrical protein chip allows for a fast detection of Staphylococcus enterotoxin B (SEB) of S. aureus and immunodominant antigen A homologue (IsaA homologue) of S. epidermidis in different liquid matrices. The S. aureus SEB virulence factor could be detected in minimal medium, milk, and urine in a concentration of 1 ng/ml within less than 23 min. Furthermore, a simultaneous detection of SEB of S. aureus and IsaA homologue of S. epidermidis in a single assay could be demonstrated.

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References

  • Albers J, Grunwald T, Nebling E, Piechotta G, Hintsche R (2003) Electrical biochip technology—a tool for microarrays and continuous monitoring. Anal Bioanal Chem 377:521–527

    Article  CAS  Google Scholar 

  • Alefantis T, Grewal P, Ashton J, Khan AS, Valdes JJ, Del Vecchio VG (2004) A rapid and sensitive magnetic bead-based immunoassay for the detection of staphylococcal enterotoxin B for high-through put screening. Mol Cell Prob 18:379–382

    Article  CAS  Google Scholar 

  • Alladyce RA, Hill AL, Murdoch DR (2006) The rapid evaluation of bacterial growth and antibiotic susceptibility in blood cultures by selected ion flow tube mass spectrometry. Diagn Microbiol Infect Dis 55:255–261

    Article  Google Scholar 

  • Archer GL (1998) Staphylococcus aureus: a well-armed pathogen. Clin Infect Dis 26(5):1179–1181

    Article  CAS  Google Scholar 

  • Asao T, Kumeda Y, Kawai T, Shibata T, Oda H, Haruki K, Nakazawa H, Kozaki S (2003) An extensive outbreak of staphylococcal food poisoning due to low-fat milk in Japan: estimation of enterotoxin A in the incriminated milk and powdered skim milk. Epidemiol Infect 130:33–40

    Article  CAS  Google Scholar 

  • Balaban N, Roosley A (2000) Staphylococcal enterotoxins. Int J Food Microbiol 61:1–10

    Article  CAS  Google Scholar 

  • Bryan CS, Kirckhart B, Brenner ER (1984) Staphylococcal bacteraemia: current patterns in nonuniversity hospitals. South Med J 77:693–696

    Article  CAS  Google Scholar 

  • Campbell GA, Medina MB, Mutharasan R (2007) Detection of Staphylococcus enterotoxin B at picogram levels using piezoelectric-excited millimeter-sized cantilever sensors. Sens Act B 126:354–360

    Article  Google Scholar 

  • Crossley KB, Archer GL (1997) The staphylococci in human disease. Churchill Livingstone, New York

    Google Scholar 

  • Elsholz B, Wörl R, Blohm L, Albers J, Feucht H, Grunwald T, Jürgen B, Schweder T, Hintsche R (2006) Automated detection and quantitation of bacterial RNA by using electrical microarrays. Anal Chem 78:4794–4802

    Article  CAS  Google Scholar 

  • Elsholz B, Nitsche A, Achenbach J, Ellerbrock H, Blohm L, Albers J, Pauli G, Hintsche R, Wörl R (2009) Electrical microarrays for highly sensitive detection of multiplex PCR products from biological agents. Biosens Bioelectron 24:1737–1743

    Article  CAS  Google Scholar 

  • Freeman J, Epstein MF, Smith NE, Platt R, Sidebottom DG, Goldman DA (1990) Extra hospital stay and antibiotic usage with nosocomial coagulase-negative staphylococcal bacteraemia in two neonatal intensive care unit populations. Am J Dis Child 144:324–329

    CAS  Google Scholar 

  • Gabig-Ciminska M, Holmgren A, Andresen H, Barken KB, Wümpelmann M, Albers J, Hintsche R, Breitenstein A, Neubauer P, Los M, Czyz A, Wegrzyn G, Silfversparre G, Jürgen B, Schweder T, Enfors SO (2004) Electric chips for rapid detection and quantification of nucleic acids. Biosens Bioelectron 19:537–546

    Article  CAS  Google Scholar 

  • Giletto A, Fyffe JG (1998) A novel ELISA format for the rapid and sensitive detection of staphylococcal enterotoxin A. Biosci Biotechnol Biochem 62(11):2217–2222

    Article  CAS  Google Scholar 

  • Gill SR, Fouts DE, Archer GL, Mongodin EF, Deboy RT, Ravel J, Paulsen IT, Kolonay JF, Brinkac L, Beanan M, Dodson RJ, Daugherty SC, Madupu R, Angiuoli SV, Durkin AS, Haft DH, Vamathevan J, Khouri H, Utterback T, Lee C, Dimitrov G, Jiang L, Qin H, Weidman J, Tran K, Kang K, Hance IR, Nelson KE, Fraser CM (2005) Insights on evolution of virulence and resistance from the complete genome analysis of an early methicillin-resistant Staphylococcus aureus strain and a biofilm-producing methicillin-resistant Staphylococcus epidermidis strain. J Bacteriol 7:2426–2438

    Article  Google Scholar 

  • Hwang SY, Kim SH, Jang EJ, Kwon NH, Park YK, Koo HC, Jung WK, Kim JM, Park YH (2007) Novel multiplex PCR for the detection of the Staphylococcus aureus superantigen and its application to raw mest isolates in Korea. Int J Food Microbiol 117(1):99–105

    Article  CAS  Google Scholar 

  • Jay JM (2000) Modern food microbiology, 6th edn. APAC, Singapore, pp 445–453

    Google Scholar 

  • Jürgen B, Barken KB, Tobisch S, Pioch D, Wümpelmann M, Hecker M, Schweder T (2005) Application of an electric DNA-chip for the expression analysis of bioprocess-relevant marker genes of Bacillus subtilis. Biotechnol Bioeng 92:299–307

    Article  Google Scholar 

  • Khan AS, Cao CJ, Thompson RG, Valdes JJ (2003) A simple and rapid fluorescence-based immunoassay for the detection of staphylococcal enterotoxin B. Mol Cell Prob 17:125–126

    Article  CAS  Google Scholar 

  • Khreich N, Lamourette P, Boutal H, Devilliers K, Creminon C, Volland H (2008) Detection of Staphylococcus enterotoxin B using fluorescent immunoliposomes as label for immunochromatographic testing. Anal Biochem 377:182–188

    Article  CAS  Google Scholar 

  • Lin HC, Tsai WC (2003) Piezoelectric crystal immunosensor for the detection of Staphylococcal enterotoxin B. Biosens Bioelectron 18:1479–1483

    Article  CAS  Google Scholar 

  • Lorenz U, Ohlsen K, Karch H, Hecker M, Thiede A, Hacker J (2000) Human antibody response during sepsis against targets expressed by methicillin resistant Staphylococcus aureus. FEMS Immunol Med Microbiol 29:145–153

    Article  CAS  Google Scholar 

  • Louie L, Majury A, Goodfellow J, Louie M (2001) Evaluation of a latex agglutination test (MRSA-Screen) for detection of oxacillin resistance in coagulase-negative staphylococci. J Cli Microbiol 39(11):4149–4151

    Article  CAS  Google Scholar 

  • Martin MC, Gonzalez-Hevia MA, Mendoza MC (2003) Usefulness of a two-step PCR procedure for detection and identification of enterotoxigenic staphylococci of bacterial isolates and food samples. Food Microbiol 20:605–610

    Article  CAS  Google Scholar 

  • Mc Cormick JK, Yarwood JM, Schlievert PM (2001) Toxic shock syndrome and bacterial superantigens: an update. Annu Rev Microbiol 55:77–104

    Article  CAS  Google Scholar 

  • Mc Lauchlin J, Narayananan GL, Mithani V, O’Neill G (2000) The detection of enterotoxins and toxic shock syndrome toxin genes in Staphylococcus aureus by polymerase chain reaction. J Food Prot 63(4):479–488

    CAS  Google Scholar 

  • Medina MB (2006) Development of a fluorescent latex microparticle immunoassay for the detection of staphylococcal enterotoxin B (SEB). J Agric Food Chem 54:4937–4942

    Article  CAS  Google Scholar 

  • Muraille E, De Smedt T, Andris F, Pajak B, Armant M, Urbain J, Moser M, Leo O (1997) Staphylococcal enterotoxin B induces an early and transient state of immunosuppression characterized by V beta-unrestricted T cell unresponsiveness and defective antigen-presenting cell functions. J Immunol 158:2638–2647

    CAS  Google Scholar 

  • Niwa O, Morita M, Tabei H (1990) Electrochemical behavior of reversible redox species at interdigitated array electrodes with different geometries: consideration of redox cycling and collection efficiency. Anal Chem 62(5):447–452

    Article  CAS  Google Scholar 

  • Novick RP (1967) Penicillinase plasmids of Staphylococcus aureus. Fed Proc 26:29–38

    CAS  Google Scholar 

  • Ohlsen K, Lorenz U (2007) Novel targets for antibiotics in Staphylococcus aureus. Future Microbiol 2:655–666

    Article  CAS  Google Scholar 

  • Paeschke M, Wollenberger U, Köhler C, Lisec T, Schnakenberg U, Hintsche R (1995) Properties of interdigital electrode arrays with different geometries. Anal Chim Acta 305:126–136

    Article  CAS  Google Scholar 

  • Palomares C, Torres MJ, Torres A, Aznar J, Polamares JC (2003) Rapid detection and identification of Staphylococcus aureus from blood culture speciemens using real time fluorescence PCR. Diagn Microbiol Infect Dis 45:183–189

    Article  CAS  Google Scholar 

  • Park CE, Warburton D, Laffey PJ, Collaberators (1996) A collaborative study on the detection of Staphylococcal enterotoxins in food with an enzyme immunoassay Kit (TECRA). J Food Protect 59(4):390–397

    CAS  Google Scholar 

  • Peruski AH, Johnsonn LH III, Peruski LF Jr (2002) Rapid and sensitive detection of biological warfare agents using time-resolved fluorescence assays. J Immunol Method 263:35–41

    Article  CAS  Google Scholar 

  • Petti CA, Fowler VG (2003) Staphylococcus aureus bacteremia and endocarditis. Cardiol Clin 2:219–233

    Article  Google Scholar 

  • Pioch D, Jürgen B, Evers S, Maurer KH, Hecker M, Schweder T (2008) Improved sandwich-hybridization assay for an electrical DNA-chip-based monitoring of bioprocess-relevant marker genes. Appl Microbiol Biotechnol 78:719–728

    Article  CAS  Google Scholar 

  • Proft T, Fraser JD (1998) Superantigens: just like peptides only different. J Exp Med 187(6):819–821

    Article  CAS  Google Scholar 

  • Proft T, Fraser JD (2007) Streptococcal superantigens. Chem Immunol Allergy 93:1–23

    Article  CAS  Google Scholar 

  • Rucker VC, Havenstrite KL, Herr AE (2005) Antibody microarrays for native toxin detection. Anal Biochem 339:262–270

    Article  CAS  Google Scholar 

  • Sabet NS, Subramaniam G, Navaratnam P, Sekaran SD (2007) Detection of methicillin- and aminoglycoside-resistant genes and simultaneous identification of S. aureus using triplex real-time PCR Taqman assay. J Microbiol Method 68:157–162

    Article  CAS  Google Scholar 

  • Sellman BR, Howell AP, Kelly-Boyd C, Baker SM (2005) Identification of immunogenic and serum binding proteins of Staphylococcus epidermidis. Infect & Immun 73(10):6591–6600

    Article  CAS  Google Scholar 

  • Shafer WM, Iandolo JJ (1979) Genetics of staphylococcal enterotoxin B in methicillin-resistant isolates of Staphylococcus aureus. Infect Immun 25:902–11

    CAS  Google Scholar 

  • Soloaga R, Corso A, Gagetti P, Faccone D, Galas M (2004) Methicillin resistance detection in Staphylococcus aureus: comparison between conventional methods and MRSA-screen latex agglutination technique. Rev Argent Microbiol 36(1):36–40

    CAS  Google Scholar 

  • Stuhlmeier R, Stuhlmeier KM (2003) Fast, simultaneous, and sensitive detection of staphylococci. J Clin Pathol 56:782–785

    Article  CAS  Google Scholar 

  • Summers WC, Brookings ES, Waites KB (1998) Identification of oxacillin-susceptible and oxacillin-resistant Staphylococcus aureus using commercial latex agglutination tests. Diagn Microbiol Infect Dis 30:131–134

    Article  CAS  Google Scholar 

  • Tenover FC, Gorwitz RJ (2006) The Epidemiology of Staphylococcus infections. In: Fischetti VA, Novick RP, Ferretti JJ (eds) Gram positive pathogens. ASM, Washington, pp 526–534

    Google Scholar 

  • Todd EC (1997) Epidemiology of foodborne diseases: a worldwide review. World Health Stat Q 50:30–50

    CAS  Google Scholar 

  • Vuong C, Otto M (2002) Staphylococcus epidermidis infections. Microbes Infect 4(4):481–489

    Article  Google Scholar 

  • Wellinghausen N, Wirths B, Essig A, Wassill L (2004) Evaluation of the hyplex bloodscreen multiplex PCR-linked immunosorbent assay system for direct identification of gram-positive cocci and gram-negative bacilli from positive blood cultures. J Clin Microbiol 42(7):3147–3152

    Article  CAS  Google Scholar 

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Acknowledgments

This work was financed by the European Union “eBIOSENSE” Electrical Bio Sensor Arrays for Analyses of Harmful Microorganisms and Microbial Toxins (no. 512009). We thank all co-workers of AJ eBiochip GmbH for their support with the eMicroLISA biosensor.

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Authors and Affiliations

  1. Pharmazeutische Biotechnologie, Institut für Pharmazie, Ernst-Moritz-Arndt-Universität, Friedrich-Ludwig-Jahn-Str. 17, 17487, Greifswald, Germany

    Annett Quiel, Britta Jürgen, Daniela Köster & Thomas Schweder

  2. Fraunhofer Institut für Siliziumtechnologie, Fraunhoferstr. 1, 25524, Itzehoe, Germany

    Gundula Piechotta

  3. Institut Scientifique d’Hygiène et d’Analyse (ISHA), Rue du Chemin Blanc-Champlan, 91165, Longjumeau, France

    Anne-Pascale Le Foll

  4. Institut für Mikrobiologie und Molekularbiologie, Ernst-Moritz-Arndt-Universität, Friedrich-Ludwig-Jahn-Str. 15, 17487, Greifswald, Germany

    Anne-Kathrin Ziebandt, Christian Kohler, Susanne Engelmann & Michael Hecker

  5. Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124, Braunschweig, Germany

    Christian Erck & Jürgen Wehland

  6. AJ eBiochip GmbH, Fraunhoferstr. 1, 25524, Itzehoe, Germany

    Rainer Hintsche

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  1. Annett Quiel
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  2. Britta Jürgen
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  4. Anne-Pascale Le Foll
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  6. Christian Kohler
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  12. Michael Hecker
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Correspondence to Thomas Schweder.

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Quiel, A., Jürgen, B., Piechotta, G. et al. Electrical protein array chips for the detection of staphylococcal virulence factors. Appl Microbiol Biotechnol 85, 1619–1627 (2010). https://doi.org/10.1007/s00253-009-2347-3

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  • DOI: https://doi.org/10.1007/s00253-009-2347-3

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