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Impact of processing methods on urinary biomarkers analysis in neonates

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Abstract

Background

In neonates, the validation of urinary biomarkers to diagnose acute kidney injury is a rapidly evolving field. The neonatal population poses unique challenges when assessing the collection, storage, and processing of urinary samples for biomarker analysis. Given this, establishing optimal and consistent sample processing in this population for meaningful use in ongoing clinical trials is important.

Methods

Urine from a cohort of 19 hospitalized neonatal intensive care unit patients enrolled in the Preterm Erythropoietin Neuroprotection Trial (Clinical Trial NCT01378273) was collected for biomarker analysis by indirect techniques using Fisher-brand cotton balls placed in the diapers. Fourteen urinary biomarkers were measured using commercially available kits via electrochemiluminescence on multiarray plates and compared between paired samples processed with centrifugation prior to storage versus prior to analysis.

Results

None of the biomarker concentrations differed between samples undergoing centrifugation prior to storage versus prior to analysis. The difference between samples was within 2% of the estimated concentration for the protein in 12 of 14 biomarkers (86%), and all paired biomarker concentrations were within 4%. The percentage error analysis did not show a difference between paired samples, with biomarker percentage errors smaller than the stated immunoassay coefficient of variance.

Conclusions

The urinary concentrations of biomarkers were comparable between paired samples, demonstrating that indirectly collected neonatal urine samples do not require centrifugation after collection and before storage. The ability to use routine urine collection and storage methods to obtain samples for subsequent quantitative immunoassay analysis should facilitate studies of newborns and young children.

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Acknowledgments

We thank Elizabeth Howland, Amy Silvia, Stephanie Hauge, and Emily Pao for their assistance with this work. This study was supported by an internal grant from the Seattle Children’s Research Institute Center for Clinical and Translational Research (SH). It was also supported by the NIH R01 DK103608 (SH, DA, SG), U01NS077953 and NCT01378273 (SJ, DM), and T32DK007662 (MS). The authors declare that they have no other relevant financial interests.

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Authors and Affiliations

  1. Division of Nephrology, Department of Pediatrics, Seattle Children’s Hospital and University of Washington, 4800 Sand Point Way NE, OC.9.830, Seattle, WA, 98105, USA

    Michelle C. Starr & Sangeeta R. Hingorani

  2. Division of Pediatric Nephrology, University of Alabama at Birmingham, Birmingham, AL, USA

    David J. Askenazi

  3. Center for Acute Care Nephrology, University of Cincinnati College of Medicine, Cincinnati, OH, USA

    Stuart L. Goldstein

  4. Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, WA, USA

    James W. MacDonald, Theo K. Bammler & Zahra Afsharinejad

  5. Department of Pediatrics, University of Iowa, Carver College of Medicine, Iowa City, IA, USA

    Patrick D. Brophy

  6. Division of Neonatology, Department of Pediatrics, University of Washington, Seattle, WA, USA

    Sandra E. Juul & Dennis E. Mayock

Authors
  1. Michelle C. Starr
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  2. David J. Askenazi
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  3. Stuart L. Goldstein
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  4. James W. MacDonald
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  5. Theo K. Bammler
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  6. Zahra Afsharinejad
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  7. Patrick D. Brophy
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  8. Sandra E. Juul
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  10. Sangeeta R. Hingorani
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Contributions

Research idea and study design: DA, SG, JM, PB, SJ, DM, SH; data acquisition: SJ, DM, SH; data analysis and interpretation: MS, DA, SG, JM, TB, ZA, PB, SH; statistical analysis: MS, JM, TB; supervision or mentorship: SH. Each author contributed important intellectual content during manuscript drafting or revision and accepts accountability for the overall work by ensuring that questions pertaining to the accuracy or integrity of any portion of the work are appropriately investigated and resolved. SH takes responsibility that this study is reported honestly, accurately, and transparently; that no important aspects of the study have been omitted; and that any discrepancies from the study as planned (and, if relevant, registered) have been explained.

Corresponding author

Correspondence to Michelle C. Starr.

Ethics declarations

Informed consent was obtained as part of the PENUT study, and all study procedures were performed in accordance with ethical standards of the local Institutional Review Board and Good Clinical Practice guidelines.

Conflict of interest

None to declare.

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Starr, M.C., Askenazi, D.J., Goldstein, S.L. et al. Impact of processing methods on urinary biomarkers analysis in neonates. Pediatr Nephrol 33, 181–186 (2018). https://doi.org/10.1007/s00467-017-3779-0

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  • DOI: https://doi.org/10.1007/s00467-017-3779-0

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