通过数字PCR测量子痫前期和正常胎盘看家基因绝对拷贝数的稳定性。
Stability of absolute copy number of housekeeping genes in preeclamptic and normal placentas, as measured by digital PCR.
作者信息
Kaitu'u-Lino T J, Hastie R, Cannon P, Lee S, Stock O, Hannan N J, Hiscock R, Tong S
机构信息
Translational Obstetrics Group, The Department of Obstetrics and Gynaecology, Mercy Hospital for Women, University of Melbourne, Heidelberg, Victoria, Australia.
Translational Obstetrics Group, The Department of Obstetrics and Gynaecology, Mercy Hospital for Women, University of Melbourne, Heidelberg, Victoria, Australia.
出版信息
Placenta. 2014 Dec;35(12):1106-9. doi: 10.1016/j.placenta.2014.10.003. Epub 2014 Oct 16.
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.
测量mRNA表达是胎盘研究的基础。理想情况下，mRNA转录本数量可直接定量。然而，使用ΔΔCT方法的PCR分析依赖管家基因的稳定性，且仅报告相对表达量。数字PCR（dPCR）可直接定量mRNA拷贝数，比定量PCR更准确。我们使用dPCR对正常血压早产（n = 20）、重度子痫前期（n = 11）和足月（n = 12）胎盘的管家基因绝对mRNA拷贝数进行了定量。虽然存在一些差异，但我们证实，在这些队列之间或整个孕期，胎盘中GAPDH、TOP1、CYC1和YWHAZ的绝对mRNA拷贝数没有显著变化。
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