Measuring fetal brain and lung transcripts in amniotic fluid supernatant: a comparison of digital PCR and RT-qPCR methods.
Purpose: Amniotic fluid (AF) cell-free RNA is a promising source of information regarding fetal physiology. Digital PCR (dPCR) is a direct approach to nucleic acid detection that reports absolute transcript copy number. The aim of this study was to compare quantification of cell-free fetal brain and...
| Publicado en: | Journal of Maternal-Fetal & Neonatal Medicine Vol. 31; no. 23; pp. 3191 - 3197 |
|---|---|
| Autores principales: | , , |
| Formato: | research Journal Article |
| Publicado: |
Taylor & Francis Ltd
Dec2018
|
| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=131688108&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 131688108 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 14767058 O6A jtl: Journal of Maternal-Fetal & Neonatal Medicine issn: 14767058 maglogo: Y pubinfo: dt: Dec2018 vid: 31 iid: 23 pid: 377 pub: Taylor & Francis Ltd place: Philadelphia, Pennsylvania artinfo: ui: 131688108 131688108 NLM28805106 131688108 10.1080/14767058.2017.1367378 NLM28805106 131688108 ppf: 3191 ppct: 6 formats: tig: atl: Measuring fetal brain and lung transcripts in amniotic fluid supernatant: a comparison of digital PCR and RT-qPCR methods. aug: au: Hui, Lisa Beard, Sally Hannan, Natalie J. affil: Mercy Perinatal, Mercy Hospital for Women, Heidelberg, Australia sug: subj: Reverse Transcriptase Polymerase Chain Reaction Amniotic Fluid Polymerase Chain Reaction Infant, Newborn Amniocentesis Pregnancy Female Genes Human Male Brain Physiology Lung Physiology Prospective Studies Validation Studies Comparative Studies Evaluation Research Multicenter Studies Infant, Newborn: birth-1 month Female Male ab: Purpose: Amniotic fluid (AF) cell-free RNA is a promising source of information regarding fetal physiology. Digital PCR (dPCR) is a direct approach to nucleic acid detection that reports absolute transcript copy number. The aim of this study was to compare quantification of cell-free fetal brain and lung RNA transcripts in AF by reverse transcription-qPCR (RT-qPCR) and dPCR.Material and Methods: Prospective hospital-based study was performed in 2016-2017. Pulmonary genes were quantified in term AF samples collected at elective cesarean birth; neurodevelopmental genes were measured in preterm samples (<34 weeks) obtained from women undergoing clinically-indicated amniocentesis.Results: All 11 women in the term cohort had three lung transcripts and a reference gene successfully amplified from their AF supernatant using RT-qPCR and dPCR. SFTPC was the most abundant lung transcript, present in higher concentrations than the reference gene in seven of the eleven samples. Neurodevelopmental gene transcripts in 12 preterm pregnancies were less reliably detected by both methods and were present in low copy numbers (<10 copies/μl). We observed significant positive correlations between transcript quantification by RT-qPCR and dPCR.Conclusion: This study confirms the presence of several potential mRNA markers of lung and brain development with dPCR and RT-qPCR, and a high correlation between the two methods. Transcripts of presumed fetal brain origin are present in very low copy numbers, which presents challenges to their feasibility as biomarkers of neurodevelopment. pubtype: Academic Journal doctype: research Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
|---|