title: mRNA heptaplex protocol for distinguishing between menstrual and peripheral blood.
abstract: The identification of menstrual blood is an important issue in forensic biology, but currently, there are no confirmatory methods for its detection. Here, we demonstrate a highly reliable simple heptaplex method that allows for the discrimination between menstrual and peripheral blood. The test has been used successfully in criminal casework, in which the origin of blood on a rape victim's underwear and trousers was questioned as being menstrual or traumatic peripheral blood. To solve this problem, transcripts of the following genes were used: mucin 4 (MUC4), human β-defensin 1 (HBD1), two matrix metalloproteinases (MMP7, MMP11), δ-aminolevulinate synthase 2 (ALAS2), hemoglobin alpha (HBA) and glucose 6-phosphate dehydrogenase (G6PDH). The sensitivity of the test is 0.3ng of RNA. The possibility of the detection and differentiation of menstrual and peripheral blood in mixtures that contain other body fluids was investigated. Reliable detection is possible for menstrual blood stains that are up to 1-2 years old if stored at room temperature. This easy approach, thanks to the amplification of 4 vaginal and 2 blood markers, minimizes the risk of false negative results.
authors.affiliation: Institute of Forensic Medicine, Medical University of Gdańsk, Dębowa 23, 80-204 Gdańsk, Poland; Department of Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdańsk and Medical University of Gdańsk, Kładki 24, 80-822 Gdańsk, Poland. Electronic address: jjakubowska@gumed.edu.pl.
authors.forename: Joanna
authors.initials: J
authors.lastname: Jakubowska
authors.affiliation: Institute of Forensic Medicine, Medical University of Gdańsk, Dębowa 23, 80-204 Gdańsk, Poland. Electronic address: amaciej@gumed.edu.pl.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Maciejewska
authors.affiliation: Department of Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdańsk and Medical University of Gdańsk, Kładki 24, 80-822 Gdańsk, Poland. Electronic address: bielawski@biotech.ug.edu.pl.
authors.forename: Krzysztof P
authors.initials: KP
authors.lastname: Bielawski
authors.affiliation: Institute of Forensic Medicine, Medical University of Gdańsk, Dębowa 23, 80-204 Gdańsk, Poland. Electronic address: richard@gumed.edu.pl.
authors.forename: Ryszard
authors.initials: R
authors.lastname: Pawłowski
mesh_terms: D000624:5-Aminolevulinate Synthetase / Q000235:genetics; D000328:Adult; D001801:Blood Stains; D005260:Female; D053803:Forensic Genetics; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics; D006454:Hemoglobins / Q000235:genetics; D006801:Humans; D053507:Matrix Metalloproteinase 11 / Q000235:genetics; D020783:Matrix Metalloproteinase 7 / Q000235:genetics; D008598:Menstruation / Q000097:blood*; D008875:Middle Aged; D055264:Mucin-4 / Q000235:genetics; D060885:Multiplex Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism*; D023083:beta-Defensins / Q000235:genetics
chemical_list: D006454:Hemoglobins; D055264:Mucin-4; D012333:RNA, Messenger; D023083:beta-Defensins; D005954:Glucosephosphate Dehydrogenase; D000624:5-Aminolevulinate Synthetase; C480079:ALAS2 protein, human; C511771:MMP11 protein, human; D053507:Matrix Metalloproteinase 11; C060932:MMP7 protein, human; D020783:Matrix Metalloproteinase 7
keywords: Blood; Forensic genetics; Menstrual blood; Multiplex PCR; mRNA profiling