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Effect of preexamination conditions in a centralized-testing model of non-invasive prenatal screening

  • Chad Fibke ORCID logo , Sylvie Giroux , André Caron , Elizabeth Starks , Jeremy D.K. Parker , Lucas Swanson , Loubna Jouan , Sylvie Langlois , Guy Rouleau , François Rousseau and Aly Karsan ORCID logo EMAIL logo
Published/Copyright: November 11, 2021
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 60 Issue 2

Abstract

Objectives

Non-invasive prenatal testing requires the presence of fetal DNA in maternal plasma. Understanding how preexamination conditions affect the integrity of cell-free DNA (cfDNA) and fetal fraction (FF) are a prerequisite for test implementation. Therefore, we examined the adjusted effect that EDTA and Streck tubes have on the cfDNA quantity and FF.

Methods

A total of 3,568 maternal blood samples across Canada were collected in either EDTA, or Streck tubes, and processing metrics, maternal body mass index (BMI), gestational age and fetal karyotype and sex were recorded. Plasma samples were sequenced using two different sequencing platforms in separate laboratories. Sequencing data were processed with SeqFF to estimate FF. Linear regression and multivariate imputation by chained equations were used to estimate the adjusted effect of tube type on cfDNA and FF.

Results

We found a positive association between cfDNA quantity and blood shipment time in EDTA tubes, which is significantly reduced with the use of Streck tubes. Furthermore, we show the storage of plasma at −80 °C is associated with a 4.4% annual relative decrease in cfDNA levels. FF was not associated with collection tube type when controlling for confounding variables. However, FF was positively associated with gestational age and trisomy 21, while negatively associated with BMI, male fetus, trisomy 18, Turners syndrome and triploidy.

Conclusions

Preexamination, maternal and fetal variables are associated with cfDNA quantity and FF. The consideration of these variables in future studies may help to reduce the number of pregnant women with inconclusive tests as a result of low FF.

Keywords: aneuploidy; cfDNA; fetal fraction; non-invasive prenatal testing (NIPT); preanalytical; stabilized blood collection tube
Corresponding author: Aly Karsan, Michael Smith Genome Sciences Centre, BC Cancer Research Institute, 675 West 10th Ave, Vancouver, BC V5Z 1L3, Canada; and Department of Pathology & Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada, E-mail:
Chad Fibke and Sylvie Giroux contributed equally to this work.

Funding source: Ariosa Diagnostics

Funding source: Genome Canada

Award Identifier / Grant number: 13527

Funding source: Canadian Institutes for Health Research

Award Identifier / Grant number: GPH-129342

Funding source: Genome BC

Award Identifier / Grant number: 145PEG

Funding source: Genome Alberta

Funding source: Québec Ministère de l’Enseignement Supérieur, de la Recherche, de la Science et de la Technologie

Funding source: Santé’s Réseau de Médecine Génétique Appliquée

Funding source: The Centre de Recherche du CHU de Québec

Acknowledgments

We would like to acknowledge the help and support given by all patients, caregivers, trial staff and researchers.

  1. Research funding: Genome Canada (13527), Canadian Institutes for Health Research (GPH-129342), Genome BC (145PEG), Genome Alberta (ROU), Québec Ministère de l’Enseignement Supérieur, de la Recherche, de la Science et de la Technologie, The Fonds de Recherche Québec – Santé’s Réseau de Médecine Génétique Appliquée, The Centre de Recherche du CHU de Québec. Ariosa Diagnostics provided in-kind testing of samples.

  2. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  3. Competing interests: Authors state no conflict of interest.

  4. Informed consent: Informed consent was obtained from all individuals included in this study.

  5. Ethical approval: Local Offices of Research Ethics approved study protocols, which have been registered in clinicaltrial.gov as trial NCT01925742.

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Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2021-0652).

Received: 2021-06-07
Accepted: 2021-10-29
Published Online: 2021-11-11
Published in Print: 2022-01-27

© 2021 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2021-0652
Keywords for this article
aneuploidy; cfDNA; fetal fraction; non-invasive prenatal testing (NIPT); preanalytical; stabilized blood collection tube

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. The clinical value of assessing the inter-method bias: the lesson from prostate specific antigen measurement
  4. Mini Review
  5. Methods to reduce lipemic interference in clinical chemistry tests: a systematic review and recommendations
  6. Opinion Paper
  7. Troponin interference with special regard to macrocomplex formation
  8. Guidelines and Recommendations from Scientific Societies
  9. Use of high-sensitivity cardiac troponins in the emergency department for the early rule-in and rule-out of acute myocardial infarction without persistent ST-segment elevation (NSTEMI) in Italy
  10. Genetics and Molecular Diagnostics
  11. Effect of preexamination conditions in a centralized-testing model of non-invasive prenatal screening
  12. General Clinical Chemistry and Laboratory Medicine
  13. Comparative study of human growth hormone measurements: impact on clinical interpretation
  14. Establishing pre-analytical requirements and maximizing peptide recovery in the analytical phase for mass spectrometric quantification of amyloid-β peptides 1–42 and 1–40 in CSF
  15. Validation of the LUMIPULSE automated immunoassay for the measurement of core AD biomarkers in cerebrospinal fluid
  16. Targeted profiling of 24 sulfated and non-sulfated bile acids in urine using two-dimensional isotope dilution UHPLC-MS/MS
  17. High-resolution capillary electrophoresis for the determination of carbamylated albumin
  18. Real-time monitoring of drug laboratory test interactions: a proof of concept
  19. Afamin predicts the prevalence and incidence of nonalcoholic fatty liver disease
  20. Reference Values and Biological Variations
  21. Blood sampling frequency as a proxy for comorbidity indices when identifying patient samples for review of reference intervals
  22. Coagulation parameters in the newborn and infant – the Copenhagen Baby Heart and COMPARE studies
  23. Hematology and Coagulation
  24. Policies and practices in the field of laboratory hematology in Croatia – a current overview and call for improvement
  25. Cardiovascular Diseases
  26. Evaluation of the Atellica TnIH cardiac troponin I assay and assessment of biological equivalence
  27. Infectious Diseases
  28. Inadequate design of mutation detection panels prevents interpretation of variants of concern: results of an external quality assessment for SARS-CoV-2 variant detection
  29. Letters to the Editors
  30. The pronounced decline of anti-SARS-CoV-2 spike trimeric IgG and RBD IgG in baseline seronegative individuals six months after BNT162b2 vaccination is consistent with the need for vaccine boosters
  31. ACE polymorphism is a determinant for COVID-19 mortality in the post-vaccination era
  32. A look at the precision, sensitivity and specificity of SARS-CoV-2 RT-PCR assays through a dedicated external quality assessment round
  33. Value of hypocalcemia and thromboinflammatory biomarkers for prediction of COVID-19 severity during the second wave: were all the waves the same?
  34. Metagenomic next-generation sequencing (mNGS) confirmed a critical case of severe fever with thrombocytopenia syndrome virus (SFTSV)
  35. Biotin interference: evaluation of an updated thyroglobulin electrochemiluminescent immunoassay
  36. Evaluation of the Beckman Coulter Access Procalcitonin Assay: analytical and clinical performance
  37. Analytical performance evaluation of the new sST2 turbidimetric assay implemented in laboratory automation systems
  38. Pre-analytical recommendations and reference values for circulating calprotectin are sample type and assay dependent
  39. Congress Abstracts
  40. Annual Meeting of the Royal Belgian Society of Laboratory Medicine: “Women’s health: from puberty to menopause”
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