Analiza porównawcza typu probówek i warunków przechowywania materiału do nieinwazyjnych badań grupy krwi płodu z wolnokrążącego DNA płodowego

2018 
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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