Analytical evaluation of a fully automated immunoassay for faecal calprotectin in a paediatric setting.

INTRODUCTION: Faecal calprotectin (FC) is a routinely used marker for identifying and monitoring children with inflammatory bowel disease (IBD). This non-invasive test is useful for screening children with gastrointestinal symptoms to avoid unnecessary invasive procedures. In this study, we validated for the first time the performance of a fully automated particle-enhanced turbidimetric immunoassay (PETIA) on the VITROS® 5600 analyzer for measurement of FC in symptomatic children and adolescents. MATERIALS AND METHODS: For performance validation of the PETIA (fCAL® turbo, Bühlmann Laboratories, Switzerland) on the VITROS® 5600 analyzer (Ortho Clinical Diagnostics, USA) limit of quantitation (LoQ), linearity, precision data and calibration curve stability were defined. Additionally, 95 faecal samples were measured using the PETIA, an enzyme-linked immunosorbent assay (ELISA; fCAL®, Bühlmann Laboratories, Switzerland) and a semi-quantitative lateral flow assay (Quantum Blue Reader®, Bühlmann Laboratories, Switzerland) for agreement evaluation. RESULTS: The LoQ for calprotectin using PETIA on the VITROS® 5600 analyzer was 21 µg/g. The linearity range was 20 - 2100 µg/g and the precision study showed a total coefficient of variation (CV) between 2.3% and 8.9%. The calibration curve was stable for 4 weeks. Using the clinical samples quantifiable by PETIA, ELISA and the semi-quantitative lateral flow assay, Passing-Bablok regression analysis and Bland-Altman plots showed good agreement. CONCLUSIONS: Due to good performance characteristics and agreement with established methods, the fully automated PETIA on the routine chemistry analyzer VITROS® 5600 is a new analytical option for the rapid determination of FC.

Peritoneal transport kinetics of proteins in children on chronic peritoneal dialysis.

OBJECTIVE: This study describes a modified 4-hour peritoneal equilibration test (PET) for analyzing peritoneal transport characteristics of proteins with different molecular weights and predicting daily peritoneal protein losses in children on chronic peritoneal dialysis (PD). DESIGN: Cross-sectional study. SETTING: A single regional pediatric dialysis unit in a teaching hospital. PATIENTS: 9 stable pediatric dialysis patients; 4 were on continuous ambulatory PD, 5 were on continuous cycling PD. MAIN OUTCOME MEASURES: Serum and dialysate concentrations of IgG, albumin, beta2-microglobulin, and transferrin were determined during a PET. Changes in dialysate-to-plasma (D/P) ratios were determined hourly. Agreement between PET-derived and measured daily peritoneal protein losses was examined. RESULTS: The D/P ratio decreased with increased molecular radius (p < 0.0001). Many children had low plasma levels of IgG, albumin, and transferrin, but elevated levels of beta2-microglobulin. The D/P ratio increased linearly during the PET for all measured proteins, regardless of molecular weight. There was close correlation between 4-hour PET protein losses and 24-hour losses during routine PD. CONCLUSIONS: Proteins are lost through the peritoneum according to their size, demonstrating linear transport kinetics during a 4-hour PET. The PET-derived data predicted daily protein losses in children on chronic PD. This approach might help to eliminate inaccuracies due to incomplete dialysate collection.