Stability of absolute copy number of housekeeping genes in preeclamptic and normal placentas, as measured by digital PCR

T J Kaitu'u-Lino; R Hastie; P Cannon; S Lee; O Stock; N J Hannan; R Hiscock; S Tong

doi:10.1016/j.placenta.2014.10.003

Stability of absolute copy number of housekeeping genes in preeclamptic and normal placentas, as measured by digital PCR

Placenta. 2014 Dec;35(12):1106-9. doi: 10.1016/j.placenta.2014.10.003. Epub 2014 Oct 16.

Authors

T J Kaitu'u-Lino¹, R Hastie², P Cannon², S Lee², O Stock², N J Hannan², R Hiscock³, S Tong²

Affiliations

¹ Translational Obstetrics Group, The Department of Obstetrics and Gynaecology, Mercy Hospital for Women, University of Melbourne, Heidelberg, Victoria, Australia. Electronic address: t.klino@unimelb.edu.au.
² Translational Obstetrics Group, The Department of Obstetrics and Gynaecology, Mercy Hospital for Women, University of Melbourne, Heidelberg, Victoria, Australia.
³ Department of Anaesthesia, Mercy Hospital for Women, Heidelberg, Victoria, Australia.

PMID: 25454476
DOI: 10.1016/j.placenta.2014.10.003

Abstract

Measuring mRNA expression is fundamental to placental research. Ideally, mRNA transcript numbers are directly quantified. However, PCR analysis using the ΔΔCT method relies on the stability of housekeeping genes and only reports relative expression. Digital PCR (dPCR) directly quantifies mRNA copy number and is more accurate than quantitative PCR. We quantified absolute mRNA copy number of housekeeping genes in normotensive pre-term (n = 20), severe preeclamptic (n = 11) and term (n = 12) placenta using dPCR. Whilst there was some variation, we confirm absolute mRNA copy number of GAPDH, TOP1, CYC1 and YWHAZ in placenta does not significantly alter between these cohorts, or across gestation.

Keywords: PCR; Placenta; Preeclampsia; Pregnancy; mRNA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Female
Genes, Essential / genetics*
Humans
Placenta / metabolism*
Polymerase Chain Reaction / methods*
Pre-Eclampsia / genetics*
Pre-Eclampsia / metabolism
Pregnancy