Pre-eclampsia and Intrauterine Growth Restriction (IUGR) are pregnancy-related diseases. Pre-eclampsia is diagnosed by the presentation of proteinuria with hypertension or proteinuria alone. Diagnosis of IUGR is based on ultrasonic fetal size measurements in the first trimester. Fetal DNA and mRNA circulate in the maternal plasma and, as recently suggested, also in maternal whole blood. Altered levels of several fetal mRNAs have been associated with pre-eclampsia and IUGR. Thus, circulating fetal mRNA could be a possible biomarker for use in the pre-symptomatic diagnosis of these diseases. Through a literary study we have aimed to identify fetal mRNA transcripts expressed specifically in the placenta and/or fetus. Based on their potential suitability, -hCG, HLA-G and CRH were selected for the aim of further investigations regarding pre-symptomatic diagnosis of the mentioned conditions. We have tried to clarify this through literary and experimental work. Messenger-RNA transcripts in whole blood have been suggested as a better alternative to plasma/serum mRNA because of higher sensitivity with regards to RT-PCR amplification. We have in our pre-study shown that GAPDH mRNA can be extracted and quantified using rtRT-PCR from whole blood. Another aim of this study was to recommend a kit most suitable for RNA purification and RT-PCR amplifications. Based on our results, -hCG is the most highly recommended transcript for further studies, since high concentrations of -hCG are found during the first trimester. In our case, the “Purelink Total RNA Blood Purification” kit from Invitrogen was the most successful for the aim of total RNA extraction from whole blood.
|Uddannelser||Medicinalbiologi, (Bachelor/kandidatuddannelse) Bachelor el. kandidat|
|Udgivelsesdato||6 jun. 2008|
- birt complications
- Fetal mRNA
- Intaruterine growth restriction
- whole blood
- Featal mRNA