Abstract
Background: Growing evidence in the literature shows the clinical importance of the calprotectin assay in faeces, especially for the differential diagnosis and monitoring of patients with inflammatory bowel diseases.
Methods: We developed a time-resolved fluorimetric immunoassay for calprotectin to extend the limited measuring range of the commercially available ELISA method currently used in our laboratory. Together with the introduction of a non-enzymatic label, this new method offers the advantages of better precision and higher sensitivity.
Results: The new assay shows a dynamic measuring range extended by a factor four, which reduces the number of samples with concentrations outside the measuring range from 30% to only 4%. The value of the assay was confirmed in various patient groups suffering from active and inactive gastrointestinal diseases. We suggest that the frequent coincidence of a high calprotectin concentration with intestinal blood loss is not the consequence of mere blood loss, but can be ascribed to neutrophil infiltration and subsequent shedding into the intestinal lumen as a result of intestinal inflammation or malignancy.
Conclusion: We expect that in the near future faecal calprotectin will be used as a non-invasive routine diagnostic marker and an effective laboratory parameter to monitor patients with inflammatory bowel disease.
References
1. Steinbakk M, Naess-Andresen CF, Lingaas E, Dale I, Brandtzaeg P, Fagerhol MK. Antimicrobial actions of calcium binding leucocyte L1 protein, calprotectin. Lancet 1990; 336:763–5.10.1016/0140-6736(90)93237-JSearch in Google Scholar
2. Van den Bos C, Roth J, Koch HG, Hartmann M, Sorg C. Phosphorylation of MRP14, and S100 protein expressed during monocytic differentiation, modulates Ca(2+)-dependent translocation from cytoplasm to membranes and cytoskeleton. J Immunol 1996; 156:1247–54.10.4049/jimmunol.156.3.1247Search in Google Scholar
3. Frosch M, Strey A, Vogl T, Wulffraat NM, Kuis W, Sunderkotter C, et al. Myeloid-related proteins 8 and 14 are specifically secreted during interaction of phagocytes and activated endothelium and are useful markers for monitoring disease activity in pauciarticular-onset juvenile rheumatoid arthritis. Arthritis Rheum 2000; 43:628–37.10.1002/1529-0131(200003)43:3<628::AID-ANR20>3.0.CO;2-XSearch in Google Scholar
4. Roseth AG, Fagerhol MK, Aadland E, Schjonsby H. Assessment of the neutrophil dominating protein calprotectin in faeces. A methodologic study. Scand J Gastroenterol 1992; 27:793–8.10.3109/00365529209011186Search in Google Scholar
5. Roseth AG, Schmidt PN, Fagerhol MK. Correlation between faecal excretion of indium-111-labelled granulocytes and calprotectin, a granulocyte marker protein, in patients with inflammatory bowel disease. Scand J Gastroenterol 1999; 34:50–4.10.1080/00365529950172835Search in Google Scholar
6. Fagerhol MK. Calprotectin, a faecal marker of organic gastrointestinal abnormality. Lancet 2000; 356:1783–4.10.1016/S0140-6736(00)03224-4Search in Google Scholar
7. Tibble JA, Teahon K, Thjodleifsson B, Roseth A, Sigthorsson G, Bridger S, et al. A simple method for assessing intestinal inflammation in Crohn's disease. Gut 2000; 47:506–13.10.1136/gut.47.4.506Search in Google Scholar
8. Summerton CB, Longlands MG, Wiener K, Shreeve DR. Faecal calprotectin: a marker of inflammation throughout the intestinal tract. Eur J Gastroenterol Hepatol 2002; 14:823–45.10.1097/00042737-200208000-00005Search in Google Scholar
9. Gaya DR, Lyon TD, Duncan A, Neilly JB, Han S, Howell J, et al. Faecal calprotectin in the assessment of Crohn's disease activity. QJM 2005; 98:435–41.10.1093/qjmed/hci069Search in Google Scholar
10. Wassell J, Dolwani S, Metzner M, Losty JH, Hawthorne A. Faecal calprotectin: a new marker for Crohn's disease? Ann Clin Biochem 2004; 41:230–2.10.1258/000456304323019613Search in Google Scholar
11. Tøn H, Brandsnes Ø, Dale S, Holtlund J, Skuibina E, Schjønsby H, et al. Improved assay for faecal calprotectin. Clin Chim Acta 2000; 292:41–54.10.1016/S0009-8981(99)00206-5Search in Google Scholar
12. Van der Sluijs Veer G, Soons JW. A time-resolved fluoroimmunoassay of the IgM-rheumatoid factor. Eur J Clin Chem Clin Biochem 1992; 30:301–5.Search in Google Scholar
13. Hemmilä I. Lanthanides as probes for time-resolved fluorometric immunoassays. Scand J Clin Lab Invest 1998; 48:389–400.10.1080/00365518809085747Search in Google Scholar
14. Hemmilä I. Applications of fluorescence in immuno-assays. New York: Wiley-Interscience, 1991.Search in Google Scholar
15. Hemmilä I, Dakabu S, Mukkala V-M, Siitari H, Lövgren T. Europium as a label in time-resolved immunofluorometric assays. Anal Biochem 1984; 137:335–43.10.1016/0003-2697(84)90095-2Search in Google Scholar
16. Hessian PA, Fisher L. The heterodimeric complex of MRP-8 (S100A8) and MRP-14 (S100A9). Antibody recognition, epitope definition and implications for structure. Eur J Biochem 2001; 268:353–63.10.1046/j.1432-1033.2001.01894.xSearch in Google Scholar PubMed
17. Goebeler M, Roth J, Teigelkamp S, Sorg C. The monoclonal antibody MAC387 detects an epitope on the calcium-binding protein MRP14. J Leukoc Biol 1994; 55:259–61.10.1002/jlb.55.2.259Search in Google Scholar PubMed
18. Holtund J, Dale S, Fagerhol MK. Process for the extraction of proteins. PCT International Publication Number WO 98/20355, 14 May 1998.Search in Google Scholar
19. Hanauer SB, Meyers S. Management of Crohn's disease in adults. Am J Gastroenterol 1997; 92:559–66.Search in Google Scholar
20. Kristinsson J, Armbruster CH, Ugstad M, Kriwanek S, Nygaard K, Ton H, et al. Faecal excretion of calprotectin in colorectal cancer: relationship to tumor characteristics. Scand J Gastroenterol 2001; 36:202–7.10.1080/003655201750065979Search in Google Scholar PubMed
21. Kronborg O, Ugstad M, Fuglerud P, Johne B, Hardcastle J, Scholefield JH, et al. Faecal calprotectin levels in a high risk population for colorectal neoplasia. Gut 2000; 46:795–800.10.1136/gut.46.6.795Search in Google Scholar PubMed PubMed Central
22. Van den Bergh FA, Kolkman JJ, Russel MG, Vlaskamp RT, Vermes I. Calprotectine: een fecale marker voor diagnostiek en follow-up bij patiënten met chronische inflammatoire darmafwijkingen. Ned Tijdschr Geneesk 2003; 147:2360–5.Search in Google Scholar
23. Tibble JA, Sigthorsson G, Foster R, Sherwood R, Fagerhol M, Bjarnason I. Faecal calprotectin and faecal occult blood tests in the diagnosis of colorectal carcinoma and adenoma. Gut 2001; 49:402–8.10.1136/gut.49.3.402Search in Google Scholar PubMed PubMed Central
24. Tibble JA, Sigthorsson G, Bridger S, Fagerhol MK, Bjarnason I. Surrogate markers of intestinal inflammation are predictive of relapse in patients with inflammatory bowel disease. Gastroenterology 2000; 119:15–22.10.1053/gast.2000.8523Search in Google Scholar PubMed
25. Costa F, Mumola MG, Ceccarelli L, Bellini M, Romano MR, Sterpi C, et al. Calprotectin is a stronger predictive marker of relapse in ulcerative colitis than in Crohn's disease. Gut 2005; 54:364–8.10.1136/gut.2004.043406Search in Google Scholar PubMed PubMed Central
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