Mathijs van de Vrie1, Jeroen K Deegens1, Johan van der Vlag1, Luuk B Hilbrands2. 1. Department of Nephrology, Radboud University Nijmegen Medical Center, Nijmegen, the Netherlands. 2. Department of Nephrology, Radboud University Nijmegen Medical Center, Nijmegen, the Netherlands. Electronic address: l.hilbrands@nier.umcn.nl.
Abstract
BACKGROUND: Levels of urinary biomarkers, such as kidney injury molecule 1 (KIM-1) and neutrophil gelatinase-associated lipocalin (NGAL), can determine the risk of renal damage and predict the outcome of multiple kidney diseases, including transplant rejection. Studies of urinary biomarkers frequently use urine that has been stored frozen before analysis without taking into account the potential of biomarker degradation during storage. STUDY DESIGN: Observational study. SETTING & PARTICIPANTS: 39 kidney transplant patients visiting an outpatient clinic. PREDICTOR: Urine storage conditions (4°C and -80°C with or without protease inhibitors) and centrifugation after thawing of samples. OUTCOMES: Urinary concentrations of NGAL and KIM-1. MEASUREMENTS: NGAL and KIM-1 were measured with commercially available enzyme-linked immunosorbent assay kits. RESULTS: Urinary NGAL and KIM-1 concentrations were stable in urine up to 48 hours when stored at 4°C and up to 6 months when stored at -80°C, independent of the addition of protease inhibitors. A centrifugation step prior to biomarker measurement did not change urinary biomarker concentrations of urine stored at -80°C. LIMITATIONS: Only KIM-1 and NGAL were measured, which possibly limits the relevance of the findings when making determinations about other urinary biomarkers. CONCLUSIONS: This study recommends that urine samples collected for the assessment of KIM-1 or NGAL are immediately cooled to 4°C and subsequently frozen at -80°C within 2 days. At -80°C, both KIM-1 and NGAL concentrations are stable for at least 6 months.
BACKGROUND: Levels of urinary biomarkers, such as kidney injury molecule 1 (KIM-1) and neutrophil gelatinase-associated lipocalin (NGAL), can determine the risk of renal damage and predict the outcome of multiple kidney diseases, including transplant rejection. Studies of urinary biomarkers frequently use urine that has been stored frozen before analysis without taking into account the potential of biomarker degradation during storage. STUDY DESIGN: Observational study. SETTING & PARTICIPANTS: 39 kidney transplant patients visiting an outpatient clinic. PREDICTOR: Urine storage conditions (4°C and -80°C with or without protease inhibitors) and centrifugation after thawing of samples. OUTCOMES: Urinary concentrations of NGAL and KIM-1. MEASUREMENTS: NGAL and KIM-1 were measured with commercially available enzyme-linked immunosorbent assay kits. RESULTS: Urinary NGAL and KIM-1 concentrations were stable in urine up to 48 hours when stored at 4°C and up to 6 months when stored at -80°C, independent of the addition of protease inhibitors. A centrifugation step prior to biomarker measurement did not change urinary biomarker concentrations of urine stored at -80°C. LIMITATIONS: Only KIM-1 and NGAL were measured, which possibly limits the relevance of the findings when making determinations about other urinary biomarkers. CONCLUSIONS: This study recommends that urine samples collected for the assessment of KIM-1 or NGAL are immediately cooled to 4°C and subsequently frozen at -80°C within 2 days. At -80°C, both KIM-1 and NGAL concentrations are stable for at least 6 months.
Authors: Meredith P Schuh; Edward Nehus; Qing Ma; Christopher Haffner; Michael Bennett; Catherine D Krawczeski; Prasad Devarajan Journal: Am J Kidney Dis Date: 2015-05-29 Impact factor: 8.860
Authors: Chi-Yuan Hsu; Dawei Xie; Sushrut S Waikar; Joseph V Bonventre; Xiaoming Zhang; Venkata Sabbisetti; Theodore E Mifflin; Josef Coresh; Clarissa J Diamantidis; Jiang He; Claudia M Lora; Edgar R Miller; Robert G Nelson; Akinlolu O Ojo; Mahboob Rahman; Jeffrey R Schelling; Francis P Wilson; Paul L Kimmel; Harold I Feldman; Ramachandran S Vasan; Kathleen D Liu Journal: Kidney Int Date: 2016-10-28 Impact factor: 10.612
Authors: Adriano César Carneiro Loureiro; Gabriella Fontenele Nocrato; André Luis Lima Correia; Robson Salviano de Matos; Júlio César Chaves Nunes Filho; Elisabeth De Francesco Daher; Flávio Henrique Macedo Pinto; Ariclécio Cunha de Oliveira; Vania Marilande Ceccatto; Rodrigo Soares Fortunato; Denise Pires de Carvalho Journal: Front Physiol Date: 2022-03-17 Impact factor: 4.566
Authors: Kathleen D Liu; Edward D Siew; W Brian Reeves; Jonathan Himmelfarb; Alan S Go; Chi-Yuan Hsu; Michael R Bennett; Prasad Devarajan; T Alp Ikizler; James S Kaufman; Paul L Kimmel; Vernon M Chinchilli; Chirag R Parikh Journal: PLoS One Date: 2016-10-27 Impact factor: 3.240
Authors: Gunnar Schley; Carmen Köberle; Ekaterina Manuilova; Sandra Rutz; Christian Forster; Michael Weyand; Ivan Formentini; Rosemarie Kientsch-Engel; Kai-Uwe Eckardt; Carsten Willam Journal: PLoS One Date: 2015-12-15 Impact factor: 3.240