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Abstract
Objective: To investigate the ability of Cell-Free RNA BCT (BCT) blood collection device to stabilize fetal cellfree RNA in maternal plasma when compared to K3EDTA collection tubes. Design and methods: Blood samples were drawn from healthy pregnant donors into K3EDTA tubes and BCTs and kept at ambient temperature (22°C). Plasma was separated by centrifugation and cell-free RNA was extracted. Circulating fetalRNAs from plasma were quantified by digital droplet polymerase chain reaction (ddPCR). Results: Blood drawn into K3EDTA tubes showed a decrease in fetal mRNA concentration for human placental lactogen (hPL), β subunit of human chorionic gonadotropin (βhCG) and placenta-specific 4 (PLAC4) over three days of ex vivo incubation at 22°C. Blood drawn into BCTs, however, showed no significant change in mRNA concentrations over the same period of time. Conclusion: Cell-Free RNA BCT blood collection devicepreservescirculating fetal RNA in maternal blood for at least for three days at 22°C, which enhances the potential clinical utility ofcirculating fetal RNA in maternal blood for noninvasive prenatal diagnostic assay development.
Highlights
The presence of circulating fetal cell-free RNA in maternal blood was initially reported in 2000 by Poon and colleagues [1]
We have shown that BCT device could minimize background RNA increase caused by agitation and temperature fluctuations that can occur during blood sample storage and shipping [11]
To study the effect of storage on cell-free β subunit of human chorionic gonadotropin (βhCG) mRNA concentration in maternal blood drawn into K3EDTA tubes and BCTs, blood samples were stored at 22°C and aliquots of blood removed from each tube on days 0, 1 and 3 and processed for the quantitation by digital PCR
Summary
The presence of circulating fetal cell-free RNA in maternal blood was initially reported in 2000 by Poon and colleagues [1]. Following this discovery, many other clinically important fetal/placental cell-free mRNA molecules were detected in maternal blood [2,3]. Using realtime quantitative RT-PCR assays, Ng and colleagues have detected and quantitated the mRNAs for human placental lactogen (hPL) and the β subunit of human chorionic gonadotropin (βhCG) in maternal blood samples [4]. Quantitation of βhCG mRNA concentration in maternal blood is clinically important because it is elevated in trisomy 21 pregnancies, but drastically reduced in trisomy 18 pregnancies [5]. The above new discoveries have presented us with opportunities to develop noninvasive prenatal diagnostic tools based on fetal cell-free RNA in maternal blood
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