title: Proof of concept study to assess fetal gene expression in amniotic fluid by nanoarray PCR.
abstract: Microarray analysis of cell-free RNA in amniotic fluid (AF) supernatant has revealed differential fetal gene expression as a function of gestational age and karyotype. Once informative genes are identified, research moves to a more focused platform such as quantitative reverse transcriptase-PCR. Standardized NanoArray PCR (SNAP) is a recently developed gene profiling technology that enables the measurement of transcripts from samples containing reduced quantities or degraded nucleic acids. We used a previously developed SNAP gene panel as proof of concept to determine whether fetal functional gene expression could be ascertained from AF supernatant. RNA was extracted and converted to cDNA from 19 AF supernatant samples of euploid fetuses between 15 to 20 weeks of gestation, and transcript abundance of 21 genes was measured. Statistically significant differences in expression, as a function of advancing gestational age, were observed for 5 of 21 genes. ANXA5, GUSB, and PPIA showed decreasing gene expression over time, whereas CASC3 and ZNF264 showed increasing gene expression over time. Statistically significantly increased expression of MTOR and STAT2 was seen in female compared with male fetuses. This study demonstrates the feasibility of focused fetal gene expression analysis using SNAP technology. In the future, this technique could be optimized to examine specific genes instrumental in fetal organ system function, which could be a useful addition to prenatal care.
authors.affiliation: Division of Genetics, Department of Pediatrics, Floating Hospital for Children at Tufts Medical Center, Boston, Massachusetts, USA. lmassingham@tuftsmedicalcenter.org
authors.forename: Lauren J
authors.initials: LJ
authors.lastname: Massingham
authors.forename: Kirby L
authors.initials: KL
authors.lastname: Johnson
authors.forename: Diana W
authors.initials: DW
authors.lastname: Bianchi
authors.forename: Shermin
authors.initials: S
authors.lastname: Pei
authors.forename: Inga
authors.initials: I
authors.lastname: Peter
authors.forename: Janet M
authors.initials: JM
authors.lastname: Cowan
authors.forename: Umadevi
authors.initials: U
authors.lastname: Tantravahi
authors.forename: Tom B
authors.initials: TB
authors.lastname: Morrison
mesh_terms: D000653:Amniotic Fluid / Q000378:metabolism*; D005260:Female; D005333:Fetus / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D008168:Lung / Q000378:metabolism; D008297:Male; D046228:Microarray Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012044:Regression Analysis