title: Forensic pregnancy diagnostics with placental mRNA markers.
abstract: 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 betahCG, with pregnancy-specific expression in whole blood samples. RT-PCR detection of hPL was positive in all samples tested throughout the pregnancy, whereas betahCG 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 betahCG 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.
authors.affiliation: Department of Forensic Molecular Biology, Erasmus University Medical Center Rotterdam, Rotterdam, The Netherlands.
authors.forename: Jeanot
authors.initials: J
authors.lastname: Gauvin
authors.forename: Dmitry
authors.initials: D
authors.lastname: Zubakov
authors.forename: Joke
authors.initials: J
authors.lastname: van Rhee-Binkhorst
authors.forename: Ate
authors.initials: A
authors.lastname: Kloosterman
authors.forename: Eric
authors.initials: E
authors.lastname: Steegers
authors.forename: Manfred
authors.initials: M
authors.lastname: Kayser
mesh_terms: D001801:Blood Stains; D018997:Chorionic Gonadotropin, beta Subunit, Human / Q000097:blood* / Q000235:genetics; D005260:Female; D053803:Forensic Genetics; D005819:Genetic Markers; D005865:Gestational Age; D006801:Humans; D010928:Placental Lactogen / Q000097:blood* / Q000235:genetics; D011247:Pregnancy / Q000097:blood* / Q000235:genetics; D011264:Pregnancy Trimesters; D012333:RNA, Messenger / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D018997:Chorionic Gonadotropin, beta Subunit, Human; D005819:Genetic Markers; D012333:RNA, Messenger; D010928:Placental Lactogen