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A comparison of sample collection methods for quantifying cell-free fetal neurodevelopment transcripts in amniotic
Lisa Hui1,2, Stephen Tong3, Tu'Uhevaha J Kaitu'u-Lino3
1Translational Obstetrics Group, Department of Obstetrics and Gynaecology, Mercy Hospital for Women, University of Melbourne, 163 Studley Rd, Heidelberg, VIC, 3084, Australia. lisahui77@gmail.com.
BMC Research Notes
|July 9, 2016
Summary
Collecting amniotic fluid (AF) in Streck RNA blood collection tubes (BCT) for fetal neurodevelopment biomarker studies did not improve cell-free RNA (cfRNA) yield. Freezing AF alone is as effective as using BCT for detecting cfRNA neurodevelopment transcripts.
Area of Science:
- Biochemistry
- Genetics
- Obstetrics
Background:
- Cell-free RNA (cfRNA) in amniotic fluid (AF) offers potential biomarkers for fetal neurodevelopment.
- Quantitative reverse transcription PCR (RT-qPCR) can detect these cfRNA transcripts.
Purpose of the Study:
- To compare cell-free RNA (cfRNA) yields from amniotic fluid (AF) collected in a commercial RNA stabilization product versus traditional freezing.
- To evaluate the utility of Streck RNA blood collection tubes (BCT) for preserving cfRNA in AF.
Main Methods:
- AF samples were collected from 13 women prior to membrane rupture during Cesarean birth.
- Samples were divided between Streck RNA BCT and plain polypropylene tubes; cfRNA was extracted.
- RT-qPCR was performed for GAPDH, NRXN3, NTRK3, and ZBTB18.
Main Results:
- cfRNA transcripts NRXN3, ZBTB18, and GAPDH were successfully amplified from all samples.
- NTRK3 was not detected in any sample.
- RNA yield was significantly lower (p < 0.0001) in samples collected in Streck RNA BCT compared to frozen samples for all detected genes.
Conclusions:
- Specific cfRNA neurodevelopment transcripts are detectable in third-trimester AF.
- Using Streck RNA BCT offers no advantage for quantitative studies of AF cfRNA and results in lower yields.
