Reliable noninvasive prenatal testing by massively parallel sequencing of circulating cell-free DNA from maternal plasma processed up to 24h after venipuncture.

Author(s): Buysse K, Beulen L, Gomes I, Gilissen C, Keesmaat C, Janssen IM, Derks-Willemen JJ, de Ligt J, Feenstra I, Bekker MN, van Vugt JM, Geurts van Kessel A, Vissers LE, Faas BH

Publication: Clin Biochem, 2013, Vol. 46, Page 1783-6

PubMed ID: 23933476 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of fetal gender and delayed centrifugation on quantification of ccffDNA and diagnosis of trisomy-21 in maternal EDTA-plasma by next generation sequencing. Plasma was stored at -80 degrees C until DNA extraction using the QIAamp Circulating Nucleic Acid kit.

   Summary of Findings:

   EDTA plasma from women carrying male fetuses produced an average of 0.036% Y chromosome reads, and the authors state fetal gender was correctly assigned in all cases, regardless of delayed centrifugation. For all but one of the 4 normal specimens, the z-scores remained within the normal range (z score of -3.0 to 3.0), even after a 48 h delay in centrifugation. The remaining normal specimen had a z-score within the normal range when blood was centrifuged within 24 h, but when centrifugation was delayed by 48 h, the Z-score was -3.03 for chromosome 21. Importantly, both trisomy 21 specimens had z scores for chromosome 21 well above the normal range (5.78-18.09), regardless of the delay in centrifugation. The percentage of ccffDNA was not significantly affected by delays in centrifugation of up to 48 h.

   Biospecimens

   • Bodily Fluid - Plasma
   • Bodily Fluid - Blood

   Preservative Types

   • Frozen

   Diagnoses:

   • Pregnant

   Platform:

   Analyte	Technology Platform
   DNA	Next generation sequencing

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Time at room temperature	6 h 24 h 48 h
   Preaquisition	Diagnosis/ patient condition	Pregnant with normal fetus Pregnant with trisomy-21 fetus
   Preaquisition	Patient gender	Female Male
   Biospecimen Aliquots and Components	Centrifugation	Centrifugation delays investigated

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