Time-resolved fluorimetric immunoassay of calprotectin: technical and clinical aspects in diagnosis of inflammatory bowel diseases

Veer, G. van der Sluijs; Hoven, Barbara van den; Rüssel, Maurice G.; Bergh, Frank A.J.T.M. van den

doi:10.1515/cclm.2006.051

Cited by 14 publications

(7 citation statements)

References 15 publications

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“…Calprotectin is a calcium binding protein of neutrophil granulocytes that correlates well with neutrophil infiltration of the intestinal mucosa when measured in feces, has antimicrobial activity, and is resistant to enzymatic degradation both in vivo and in vitro [9,40]. Accordingly, as a marker of neutrophilic intestinal inflammation, calprotectin values might reflect a composite endpoint for organic intestinal disease.…”

Section: Discussionmentioning

confidence: 99%

Value of fecal calprotectin in the evaluation of patients with abdominal discomfort: an observational study

Manz

Burri

Rothen³

et al. 2012

BMC Gastroenterol

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BackgroundThe evaluation of patients with abdominal discomfort is challenging and patient selection for endoscopy based on symptoms is not reliable. We evaluated the diagnostic value of fecal calprotectin in patients with abdominal discomfort.MethodsIn an observational study, 575 consecutive patients with abdominal discomfort referred for endoscopy to the Department of Gastroenterology & Hepatology at the University Hospital Basel in Switzerland, were enrolled in the study. Calprotectin was measured in stool samples collected within 24 hours before the investigation using an enzyme-linked immunosorbent assay. The presence of a clinically significant finding in the gastrointestinal tract was the primary endpoint of the study. Final diagnoses were adjudicated blinded to calprotectin values.ResultsMedian calprotectin levels were higher in patients with significant findings (N = 212, median 97 μg/g, IQR 43-185) than in patients without (N = 326, 10 μg/g, IQR 10-23, P < 0.001). The area under the receiver operating characteristics curve (AUC) to identify a significant finding was 0.877 (95% CI, 0.85-0.90). Using 50 μg/g as cut off yielded a sensitivity of 73% and a specificity of 93% with good positive and negative likelihood ratios (10.8 and 0.29, respectively). Fecal calprotectin was useful as a diagnostic parameter both for findings in the upper intestinal tract (AUC 0.730, 0.66-0.79) and for the colon (AUC 0.912, 0.88-0.94) with higher diagnostic precision for the latter (P < 0.001). In patients > 50 years, the diagnostic precision remained unchanged (AUC 0.889 vs. 0.832, P = 0.165).ConclusionIn patients with abdominal discomfort, fecal calprotectin is a useful non-invasive marker to identify clinically significant findings of the gastrointestinal tract, irrespective of age.

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Section: Discussionmentioning

confidence: 99%

Value of fecal calprotectin in the evaluation of patients with abdominal discomfort: an observational study

Manz

Burri

Rothen³

et al. 2012

BMC Gastroenterol

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Section: Methodsmentioning

confidence: 99%

“…A total of 100 mg faeces was weighed into a 15 mL tube, and 4.9 mL extraction buffer (0.1 M Tris, 0.15 M NaCl, 1.0 M urea, 10 mM CaCl 2 ·2H 2 O, 0.1 M citric acid, and 0.5% bovine serum albumin, pH 8.0) was added. 21 After 90 min of mixing, 1 mL of suspension was centrifuged at 10 000 g for 5 min at 4°C, and 700 μl supernatant was transferred into a fresh tube and stored at −80°C. Calprotectin concentrations were measured using a commercially available human faecal calprotectin enzyme‐linked immunosorbent assay (lower detection limit 2.56 μg/g) (Hycult Biotech, Uden, the Netherlands).…”

Section: Methodsmentioning

confidence: 99%

Gut microbiota and short‐chain fatty acid alterations in cachectic cancer patients

Ubachs

Ziemons

Soons

et al. 2021

J cachexia sarcopenia muscle

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Background Cancer cachexia is characterized by a negative energy balance, muscle and adipose tissue wasting, insulin resistance, and systemic inflammation. Because of its strong negative impact on prognosis and its multifactorial nature that is still not fully understood, cachexia remains an important challenge in the field of cancer treatment. Recent animal studies indicate that the gut microbiota is involved in the pathogenesis and manifestation of cancer cachexia, but human data are lacking. The present study investigates gut microbiota composition, short‐chain fatty acids (SCFA), and inflammatory parameters in human cancer cachexia. Methods Faecal samples were prospectively collected in patients (N = 107) with pancreatic cancer, lung cancer, breast cancer, or ovarian cancer. Household partners (N = 76) of the patients were included as healthy controls with similar diet and environmental conditions. Patients were classified as cachectic if they lost >5% body weight in the last 6 months. Gut microbiota composition was analysed by sequencing of the 16S rRNA V4 gene region. Faecal SCFA levels were quantified by gas chromatography. Faecal calprotectin was assessed with enzyme‐linked immunosorbent assay. Serum C‐reactive protein and leucocyte counts were retrieved from medical records. Results Cachexia prevalence was highest in pancreatic cancer (66.7%), followed by ovarian cancer (25%), lung cancer (20.8%), and breast cancer (17.3%). Microbial α‐diversity was not significantly different between cachectic cancer patients (N = 33), non‐cachectic cancer patients (N = 74), or healthy controls (N = 76) (species richness P = 0.31; Shannon effective index P = 0.46). Community structure (β‐diversity) tended to differ between these groups (P = 0.053), although overall differences were subtle and no clear clustering of samples was observed. Proteobacteria (P < 0.001), an unknown genus from the Enterobacteriaceae family (P < 0.01), and Veillonella (P < 0.001) were more abundant among cachectic cancer patients. Megamonas (P < 0.05) and Peptococcus (P < 0.001) also showed differential abundance. Faecal levels of all SCFA tended to be lower in cachectic cancer patients, but only acetate concentrations were significantly reduced (P < 0.05). Faecal calprotectin levels were positively correlated with the abundance of Peptococcus, unknown Enterobacteriaceae, and Veillonella. We also identified several correlations and interactions between clinical and microbial parameters. Conclusions This clinical study provided the first insights into the alterations of gut microbiota composition and SCFA levels that occur in cachectic cancer patients and how they are related to inflammatory parameters. These results pave the way for further research examining the role of the gut microbiota in cancer cachexia and its potential use as therapeutic target.

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“…All faecal samples were collected within 7 hours after cycling. For faecal analysis, faeces were thawed, and 100 mg was added to 4.9 ml of extraction buffer (0.1 M Tris, 0.15 M NaCl, 1.0 M urea, 10 mM CaCl 2 2H 2 O, 0.1 M citric acid, 0.5% BSA, pH 8.0) was added [48]. After 30 minutes shaking, 1 ml of suspension was centrifuged at 10,000 rpm for 20 minutes at 4°C.…”

Section: Methodsmentioning

confidence: 99%

Exercise-Induced Splanchnic Hypoperfusion Results in Gut Dysfunction in Healthy Men

et al. 2011

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BackgroundSplanchnic hypoperfusion is common in various pathophysiological conditions and often considered to lead to gut dysfunction. While it is known that physiological situations such as physical exercise also result in splanchnic hypoperfusion, the consequences of flow redistribution at the expense of abdominal organs remained to be determined. This study focuses on the effects of splanchnic hypoperfusion on the gut, and the relationship between hypoperfusion, intestinal injury and permeability during physical exercise in healthy men.Methods and FindingsHealthy men cycled for 60 minutes at 70% of maximum workload capacity. Splanchnic hypoperfusion was assessed using gastric tonometry. Blood, sampled every 10 minutes, was analyzed for enterocyte damage parameters (intestinal fatty acid binding protein (I-FABP) and ileal bile acid binding protein (I-BABP)). Changes in intestinal permeability were assessed using sugar probes. Furthermore, liver and renal parameters were assessed. Splanchnic perfusion rapidly decreased during exercise, reflected by increased gapg-apCO2 from −0.85±0.15 to 0.85±0.42 kPa (p<0.001). Hypoperfusion increased plasma I-FABP (615±118 vs. 309±46 pg/ml, p<0.001) and I-BABP (14.30±2.20 vs. 5.06±1.27 ng/ml, p<0.001), and hypoperfusion correlated significantly with this small intestinal damage (rS = 0.59; p<0.001). Last of all, plasma analysis revealed an increase in small intestinal permeability after exercise (p<0.001), which correlated with intestinal injury (rS = 0.50; p<0.001). Liver parameters, but not renal parameters were elevated.ConclusionsExercise-induced splanchnic hypoperfusion results in quantifiable small intestinal injury. Importantly, the extent of intestinal injury correlates with transiently increased small intestinal permeability, indicating gut barrier dysfunction in healthy individuals. These physiological observations increase our knowledge of splanchnic hypoperfusion sequelae, and may help to understand and prevent these phenomena in patients.

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Time-resolved fluorimetric immunoassay of calprotectin: technical and clinical aspects in diagnosis of inflammatory bowel diseases

Abstract: 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.

Cited by 14 publications

References 15 publications

Value of fecal calprotectin in the evaluation of patients with abdominal discomfort: an observational study

Value of fecal calprotectin in the evaluation of patients with abdominal discomfort: an observational study

Gut microbiota and short‐chain fatty acid alterations in cachectic cancer patients

Exercise-Induced Splanchnic Hypoperfusion Results in Gut Dysfunction in Healthy Men

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