Turkiye Klinikleri Journal of Forensic Medicine and Forensic Sciences

.: ORIGINAL RESEARCH
Vücut Sıvılarının Tanımlanmasında Çeşitli Doku Spesifik mRNA Belirteçlerinin Etkinliğinin Araştırılması
An Investigation of Various Tissue Specific mRNA Markers in Body Fluid Identification
Ayşe SERİNa, Hüsniye CANANa, Ayça ULUBAYa, Behnan ALPERa
aAdli Tıp AD, Çukurova Üniversitesi Tıp Fakültesi, Adana
Turkiye Klinikleri J Foren Sci Leg Med. 2017;14(2):48-55
doi: 10.5336/forensic.2017-57328
Article Language: TR
Full Text
ÖZET
Amaç: Tam kan için hemoglobin beta (HBB), beta spektrin (SPTB), porfobilinojen deaminaz (PBGD), semen için protamin 1 (PRM1), protamin 2 (PRM2), menstrüel kan için matriks metalloproteinaz 7 (MMP7) ve 11 (MMP11) ile tükürük için statherin (STATH), histatin 3 (HTN3) ve vajinal sekresyon için mucin 4 (MUC4), insan beta defensin 1 (HBD1) mRNA'larının hedef dokuların tanımlanmasında kullanılabilirliğinin araştırılmasıdır. Çalışmada, 18S ribozomal RNA (18 SRNA) ve gliseraldehit fosfat dehidrogenaz (GAPDH) endojen kontrol genleri olarak kullanılmıştır. Gereç ve Yöntemler: Deneysel olarak hazırlanan 10?ar adet kan, menstrüel kan, semen, tükürük ve vajinal sekresyon lekesinde sırası ile RNA izolasyonu, cDNA sentezi, single- ve multipleks-polimeraz zincir reaksiyonu (PCR) gerçekleştirilmiştir. Ardından PCR ürünleri otomatik kapiller elektroforezde yürütülmüştür. Bulgular: Çalışmada kullanılan tüm mRNA belirteçleri hedef vücut sıvılarında bulunmuştur. Semen, tükürük, vajinal sekresyon belirteçleri, ilgili vücut sıvıları için spesifik iken; diğer belirteçlerde çapraz reaksiyonlar da gözlenmiştir. Kan, menstrüel kan, semen, tükürük ve vajinal sekresyon belirteçlerinin tamamı deneysel olarak hazırlanan ve farklı ortam koşullarında bekletilen 6 aylık lekelerde gösterilebilmiştir. Sonuç: Bulgular, vücut sıvılarının identifikasyonunda seçilen belirteçlerden MMP11 dışındakilerin hedef dokuların ayrımında kullanılabileceğini, ayrıca bu belirteçlerin multipleks amplifikasyonu ile lekelerin hedef vücut sıvılarına ait olup olamayacağı sorusuna bir kere de yanıt verebileceğini göstermektedir.

Anahtar Kelimeler: RNA stabilitesi; adli genetik; vücut sıvıları
ABSTRACT
Objective: The goal of this study was to investigate the usefulness of mRNAs in the identification of target tissues: hemoglobin beta (HBB), beta spectrin (SPTB), and porphobilinogen deaminase (PBGD) for whole blood; protamine 1 (PRM1) and 2 (PRM2) for semen; matrix metalloproteinase 7 (MMP-7) and 11 (MMP-11) for menstrual blood; statherin (STATH) and histatin 3 (HTN3) for saliva; and mucin 4 (MUC4) and human beta defensin 1 (hBD1) for vaginal secretion In the study 18S ribosomal RNA (18S rRNA) and glyceraldehyde phosphate dehydrogenase (GAPDH) were used as endogenous control genes. Material and Methods: RNA isolation, cDNA synthesis, and single- and multiplex PCR were performed for each of 10 experimental samples consisted of whole blood, menstrual blood, semen, saliva, and vaginal secretion. The PCR products were run on capillary electrophoresis. Results: All mRNA markers used in the study were found in the target body fluids. Semen, saliva and vaginal secretion markers were specific for the respective body fluids, while cross-reactions were also observed in the other markers. All of the whole blood, menstrual blood, semen, saliva, and vaginal secretion markers were present in 6-month stains that had been prepared experimentally and stored at various ambient conditions. Conclusion: The findings show that, with the exception of MMP-11, the selected markers for identification of aforementioned body fluids can all be used in the distinguishing of target tissues, and that with multiplex amplification these markers can give an immediate answer to the question of whether or not the stains originated to the target body fluids.

Keywords: RNA stability; forensic genetics; body fluids

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