open access

Vol 11, No 4 (2018)
Research paper
Published online: 2019-03-11
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Comparative analysis of the type of tubes and storage conditions of material for non-invasive prenatal testing (NIPT) of fetal blood group and platelet antigens

Agnieszka Orzińska, Katarzyna Guz, Sylwia Purchla-Szepioła, Magdalena Krzemienowska, Pamela Bartoszewicz, Marzena Dębska, Izabela Kopeć, Małgorzata Uhrynowska, Ewa Brojer
Journal of Transfusion Medicine 2018;11(4):137-143.

open access

Vol 11, No 4 (2018)
ORIGINAL PAPERS
Published online: 2019-03-11

Abstract

Background: The stability and proportion of fetal/maternal DNA isolated from maternal plasma are crucial for the accuracy of non-invasive prenatal testing (NIPT) of fetal blood group and platelet antigens. Aim: Comparative analysis of NIPT of material collected into tubes routinely used at the Institute of Hematology and Transfusion Medicine (IHTM) and into special tubes dedicated for cell-free (cf) DNA that stabilizes maternal blood cells. Methods: Blood from 29 pregnant RhD-negative women was collected in parallel into EDTA vacutainer tubes with gel barrier (BD Vacutainer PPT, USA) and into Cell-Free DNA Collection tubes (Roche). From the latter samples plasma was processed immediately after collection (n = 18), after 48-96h of storage in ambient temperature (n = 24) and after freezing plasma fractions separated immediately after collection (n = 27). DNA was isolated using easyMag (Biomerieux) and examined by real-time PCR on LC480II (Roche) for the presence of fetal and maternal markers. Results: In all study groups the differences in fetal fraction were not statistically significant (p > 0,05). However the maternal fraction in plasma collected into special cfDNA tubes and tested within 24h or from frozen plasma was significantly lower than from the tested standard tubes. Conclusion: The type of tubes into which maternal blood is collected does not affect the level of fetal DNA fraction. On the other hand, collection of blood into special cfDNA tubes contributes to stabilization of the level of maternal fraction provided blood is stored at ambient temperature and processed within 96h of venipuncture. Such conditions are recommended for NIPT of fetal antigens performed in IHTM.

Abstract

Background: The stability and proportion of fetal/maternal DNA isolated from maternal plasma are crucial for the accuracy of non-invasive prenatal testing (NIPT) of fetal blood group and platelet antigens. Aim: Comparative analysis of NIPT of material collected into tubes routinely used at the Institute of Hematology and Transfusion Medicine (IHTM) and into special tubes dedicated for cell-free (cf) DNA that stabilizes maternal blood cells. Methods: Blood from 29 pregnant RhD-negative women was collected in parallel into EDTA vacutainer tubes with gel barrier (BD Vacutainer PPT, USA) and into Cell-Free DNA Collection tubes (Roche). From the latter samples plasma was processed immediately after collection (n = 18), after 48-96h of storage in ambient temperature (n = 24) and after freezing plasma fractions separated immediately after collection (n = 27). DNA was isolated using easyMag (Biomerieux) and examined by real-time PCR on LC480II (Roche) for the presence of fetal and maternal markers. Results: In all study groups the differences in fetal fraction were not statistically significant (p > 0,05). However the maternal fraction in plasma collected into special cfDNA tubes and tested within 24h or from frozen plasma was significantly lower than from the tested standard tubes. Conclusion: The type of tubes into which maternal blood is collected does not affect the level of fetal DNA fraction. On the other hand, collection of blood into special cfDNA tubes contributes to stabilization of the level of maternal fraction provided blood is stored at ambient temperature and processed within 96h of venipuncture. Such conditions are recommended for NIPT of fetal antigens performed in IHTM.
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Keywords

cell-free fetal DNA; non-invasive prenatal testing; red blood cell and platelet antigens

About this article
Title

Comparative analysis of the type of tubes and storage conditions of material for non-invasive prenatal testing (NIPT) of fetal blood group and platelet antigens

Journal

Journal of Transfusion Medicine

Issue

Vol 11, No 4 (2018)

Article type

Research paper

Pages

137-143

Published online

2019-03-11

Bibliographic record

Journal of Transfusion Medicine 2018;11(4):137-143.

Keywords

cell-free fetal DNA
non-invasive prenatal testing
red blood cell and platelet antigens

Authors

Agnieszka Orzińska
Katarzyna Guz
Sylwia Purchla-Szepioła
Magdalena Krzemienowska
Pamela Bartoszewicz
Marzena Dębska
Izabela Kopeć
Małgorzata Uhrynowska
Ewa Brojer

References (13)
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