Skip to main content

Advertisement

Log in

Serological microarray for a paradoxical diagnostic of Whipple’s disease

  • Article
  • Published:
European Journal of Clinical Microbiology & Infectious Diseases Aims and scope Submit manuscript

Abstract

Whipple’s disease is a systemic chronic infection caused by Tropheryma whipplei. Asymptomatic people may carry T. whipplei in their digestive tract and this can be determined by PCR, making serological diagnosis useful to distinguish between carriers and patients. Putative antigenic proteins were selected by computational analysis of the T. whipplei genome, immunoproteomics studies and from literature. After expression, putative T. whipplei antigens were screened by microimmunofluorescence with sera of immunized rabbit. Selected targets were screened by microarray using sera from patients and carriers. Paradoxically, with 19 tested recombinant proteins and a glycosylated native protein of T. whipplei, a higher immune response was observed with asymptomatic carriers. In contrast, quantification of human IgA exhibited a higher reaction in patients than in carriers against 10 antigens. These results were used to design a diagnostic test with a cut-off value for each antigen. A blind test assay was performed and was able to diagnose 6/8 patients and 11/12 carriers. Among people with positive T. whipplei PCR of the stool, patients differ from carriers by having positive IgA detection and a negative IgG detection. If confirmed, this serological test will distinguish between carriers and patients in people with positive PCR of the stool.

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

Access this article

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

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. La Scola B, Fenollar F, Fournier PE, Altwegg M, Mallet MN, Raoult D (2001) Description of Tropheryma whipplei gen.nov., sp.nov., the Whipple’s disease bacillus. Int J Syst Evol Microbiol 51:1471–1479

    PubMed  Google Scholar 

  2. Wilson KH, Blitchington R, Frothingham R, Wilson JA (1991) Phylogeny of the WhippIe’s-disease-associated bacterium. Lancet 338:474–475

    Article  PubMed  CAS  Google Scholar 

  3. Relman DA, Schmidt TM, MacDermott RP, Falkow S (1992) Identification of the uncultured bacillus of Whipple’s disease. N Engl J Med 327:293–301

    PubMed  CAS  Google Scholar 

  4. Raoult D, Birg ML, La Scola B, Fournier PE, Enea M, Lepidi H, Roux V, Piette JC, Vandenesch F, Vital Durand D, Marrie TJ (2000) Cultivation of the bacillus of Whipple’s disease. New Engl J Med 342:620–625

    Article  PubMed  CAS  Google Scholar 

  5. Renesto P, Crapoulet N, Ogata H, La Scola B, Vestris G, Claverie JM, Raoult D (2003) Genome-based design of a cell-free culture medium for Tropheryma whipplei. Lancet 362:447–449

    Article  PubMed  Google Scholar 

  6. Bentley SD, Maiwald M, Murphy LD, Pallen MJ, Yeats CA, Dover LG, Norbertczak HT, Besra GS, Quail MA, Harris DE, Von Herbay A, Goble A, Rutter S, Squares R, Squares S, Barrell BG, Parkhill J, Relman DA (2003) Sequencing and analysis of the genome of the Whipple’s disease bacterium Tropheryma whipplei. Lancet 361:637–644

    Article  PubMed  CAS  Google Scholar 

  7. Raoult D, Ogata H, Audic S, Robert C, Suhre K, Drancourt M, Claverie JM (2003) Tropheryma whipplei twist: a human pathogenic actinobacteria with a reduced genome. Genome Res 13:1800–1809

    PubMed  CAS  Google Scholar 

  8. Kowalczewska M, Fenollar F, Lafitte D, Raoult D (2006) Identification of candidate antigen in Whipple’s disease using a serological proteomic approach. Proteomics 6:3294–3305

    Article  PubMed  CAS  Google Scholar 

  9. Fenollar F, Puechal X, Raoult D (2007) Whipple’s disease. N Engl J Med 356:55–66

    Article  PubMed  CAS  Google Scholar 

  10. Fenollar F, Raoult D (2001) Whipple’s disease. Clin Diagn Lab Immunol 8:1–8

    Article  PubMed  CAS  Google Scholar 

  11. Misbah SA, Mapstone NP (2000) Whipple’s disease revisited. J Clin Pathol 53:750–755

    Article  PubMed  CAS  Google Scholar 

  12. Maiwald M, Relman D (2001) Whipple’s disease and Tropheryma whippelii: secrets slowly revealed. Clin Infect Dis 32:457–463

    Article  PubMed  CAS  Google Scholar 

  13. Mahnel R, Marth T (2004) Progress, problems, and perspectives in diagnosis and treatment of Whipple’s disease. Clin Exp Med 4:39–43

    Article  PubMed  CAS  Google Scholar 

  14. Black-Schaffer B (1949) The tinctoral demonstration of a glycoprotein in Whipple’s disease. Proc Soc Exp Biol 72:225–227

    CAS  Google Scholar 

  15. Marth T, Raoult D (2003) Whipple’s disease. Lancet 361:239–246

    Article  PubMed  Google Scholar 

  16. Schoniger-Hekele M, Petermann D, Weber B, Muller C (2007) Tropheryma whipplei in the environment: survey of sewage plant influxes and sewage plant workers. Appl Environ Microbiol 73:2033–2035

    Article  PubMed  Google Scholar 

  17. Fenollar F, Trani M, Davoust B, Salle B, Birg ML, Rolain JM, Raoult D (2008) Carriage of Tropheryma whipplei in stools of sewer workers and human controls, but not in monkeys and apes. J Infect Dis In press

  18. Li W, Fenollar F, Rolain JM, Fournier PE, Feurle GE, Muller C, Moos A, Marth T, Altwegg M, Calligaris-Maibach R, Schneider T, Biagi F, La Scola B, Raoult D (2008) Genotyping reveals a wide heterogeneity of Tropheryma whipplei. Microbiology 154:527

    Google Scholar 

  19. Desnues B, Raoult D, Mege JL (2005) IL-16 is critical for Tropheryma whipplei replication in Whipple’s disease. J Immunol 175:4575–4582

    PubMed  CAS  Google Scholar 

  20. Desnues B, Ihrig M, Raoult D, Mege JL (2006) Whipple’s disease: a macrophage disease. Clin Vaccine Immunol 13:170–178

    Article  PubMed  CAS  Google Scholar 

  21. Benoit M, Fenollar F, Raoult D, Mege JL (2007) Increased levels of circulating IL-16 and apoptosis markers are related to the activity of Whipple’s Disease. PLoS ONE 2:e494

    Article  PubMed  Google Scholar 

  22. Kowalczewska M, Fenollar F, Villard C, Azza S, Roux M, Raoult D (2008) An immunoproteomic approach for identification of clinical biomarkers of Whipple’s disease. Proteomics Clin Appl In press

  23. Bacarese-Hamilton T, Bistoni F, Crisanti A (2002) Protein microarrays: from serodiagnosis to whole proteome scale analysis of the immune response against pathogenic microorganisms. Biotechniques Suppl:S24-S29

  24. Mezzasoma L, Bacarese-Hamilton T, Di Cristina M, Rossi R, Bistoni F, Crisanti A (2002) Antigen microarrays for serodiagnosis of infectious diseases. Clin Chem 48:121–130

    PubMed  CAS  Google Scholar 

  25. Steller S, Angenendt P, Cahill DJ, Heuberger S, Lehrach H, Kreutzberger J (2005) Bacterial protein microarrays for identification of new potential diagnostic markers for Neisseria meningitidis infections. Proteomics 5:2048–2055

    Article  PubMed  CAS  Google Scholar 

  26. Bonhomme CJ, Nappez C, Raoult D (2007) Microarray for serotyping of Bartonella species. BMC Microbiol 7:59

    Article  PubMed  Google Scholar 

  27. Gouriet F, Drancourt M, Raoult D (2006) Multiplexed serology in atypical bacterial pneumonia. Ann N Y Acad Sci 1078:530–540

    Article  PubMed  CAS  Google Scholar 

  28. Saha S, Raghava GPS (2004) BcePred: Prediction of continuous B-cell epitopes in antigenic sequences using physico-chemical properties. In: Nicosia G, Cutello V, Bentley PJ, Timmis J (eds) ICARIS, LNCS. Springer, pp 197–204

  29. Rolain JM, Lecam C, Raoult D (2003) Simplified serological diagnosis of endocarditis due to Coxiella burnetii and Bartonella. Clin Diagn Lab Immunol 10:1147–1148

    Article  PubMed  CAS  Google Scholar 

  30. Eshaghi S, Hedren M, Nasser MI, Hammarberg T, Thornell A, Nordlund P (2005) An efficient strategy for high-throughput expression screening of recombinant integral membrane proteins. Protein Sci 14:676–683

    Article  PubMed  CAS  Google Scholar 

  31. Sagara K, Matsuyama S, Mizushima S (1994) SecF stabilizes SecD and SecY, components of the protein translocation machinery of the Escherichia coli cytoplasmic membrane. J Bacteriol 176:4111–4116

    PubMed  CAS  Google Scholar 

  32. Gouriet F, Levy PY, Samson L, Drancourt M, Raoult D (2008) Comparison of multiplexed slides InoDiag automate immunofluorescence to reference serology in the diagnosis of atypical bacterial pneumonia. Clin Microbiol Infect submitted

  33. Bowie JU (2005) Solving the membrane protein folding problem. Nature 438:581–589

    Article  PubMed  CAS  Google Scholar 

  34. Walhout AJ, Temple GF, Brasch MA, Hartley JL, Lorson MA, van den Heuvel S, Vidal M (2000) GATEWAY recombinational cloning: application to the cloning of large numbers of open reading frames or ORFeomes. Methods Enzymol 328:575–592

    Article  PubMed  CAS  Google Scholar 

  35. Lepidi H, Fenollar F, Gerolami R, Mege JL, Bonzi MF, Chappuis M, Sahel J, Raoult D (2003) Whipple’s disease: Immunospecific and quantitative immunohistochemical study of intestinal biopsy specimens. Hum Pathol 34:589–596

    Article  PubMed  Google Scholar 

  36. Eshaghi S, Hedren M, Nasser MI, Hammarberg T, Thornell A, Nordlund P (2005) An efficient strategy for high-throughput expression screening of recombinant integral membrane proteins. Protein Sci 14:676–683

    Article  PubMed  CAS  Google Scholar 

  37. Kalt A, Schneider T, Ring S, Hoffmann J, Zeitz M, Stallmach A, Persing DH, Marth T (2006) Decreased levels of interleukin-12p40 in the serum of patients with Whipple’s disease. Int J Colorectal Dis 21:114–120

    Article  PubMed  CAS  Google Scholar 

  38. Mahnel R, Kalt A, Ring S, Stallmach A, Strober W, Marth T (2005) Immunosuppressive therapy in Whipple’s disease patients is associated with the appearance of gastrointestinal manifestations. Am J Gastroenterol 100:1167–1173

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgment

We thank Inodiag for technical help and for use of their technology.

We thank Isabelle Combe for helping in formatting the manuscript.

Conflicts of interest statement

Didier RAOULT is a cofounder of INODIAG, a start-up localized in LA CIOTAT France.

Financial support

This work was supported by a grant from the fifth Framework Program of the European Commission (QL G1-CT-2002–01049) and a grant for Programme Hospitalier de Recherche Clinique from the French Ministry of Health.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to D. Raoult.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bonhomme, C.J., Renesto, P., Nandi, S. et al. Serological microarray for a paradoxical diagnostic of Whipple’s disease. Eur J Clin Microbiol Infect Dis 27, 959–968 (2008). https://doi.org/10.1007/s10096-008-0528-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10096-008-0528-0

Keywords

Navigation