Current methods for pregnancy diagnostics are based on immunodetection of pregnancy-specific proteins and in a forensic context
suffer from sensitivity and specificity issues. Here, we applied reverse transcriptase polymerase chain reaction (RT-PCR)
technology to 11 genes previously reported with placental mRNA circulating in maternal blood. We found two genes,
hPL and
βhCG, with pregnancy-specific expression in whole blood samples. RT-PCR detection of
hPL was positive in all samples tested throughout the pregnancy, whereas
βhCG was detectable until half of the second trimester but not at later gestation ages. For
hPL, in vitro stability of the transcript was demonstrated until 2 months of age, and the
hPL-specific RT-PCR assay applied was highly sensitive with reliable detection from down to 0.25 cm
2 dried bloodstain. We therefore suggest
hPL-specific RT-PCR as a new molecular tool for forensic pregnancy diagnostics from dried blood stains. Moreover, our results
indicate that the time-wise reverse expression of
hPL and
βhCG during pregnancy may allow an RT-PCR-based estimation of the gestational age from blood stains, adding to the value of forensic
pregnancy diagnosis for crime scene investigations.
Keywords mRNA markers - Pregnancy - Maternal blood - Forensic -
hPL
-
βhCG
Jeanot Gauvin, Dmitry Zubakov contributed equally to this work.