IDENTIFICATION OF WORLD WAR II VICTIMS USING DNA ANALYSIS OF BONE SAMPLES

Ciljevi rada: Detaljno opisati proces identifikacije osoba analizom DNA iz koštanih uzoraka II. svjetskog rata i istaknuti probleme izolacije DNA iz starih koštanih uzoraka te probleme pronalaska uže rodbine za usporedbu DNA. Metode: Izolacija DNA iz uzoraka kosti lubanje i femura provedena je standardnom fenol/kloroform/izoamilni alkohol ekstrakcijom. Prije ekstrakcije uzorci su tretirani metodom EDTA dekalcifikacije. Kvantifikacija humane DNA napravljena je na RT-PCR ABI Prism 7000 Sequence Detection System instrumentu. PCR amplifikacija obavljena je u GeneAmp® PCR System 9700 instrumentu, a za umnažanje je korišten AmpFLSTR® Identifiler® PCR Amplification Kit. Umnoženi DNA fragmenti razdvojeni su na ABI PRISM ® 310 Genetic Analyzer instrumentu. Rezultati: Analizom koštanih uzoraka dobiven je nepotpuni DNA profil osobe. Identifikacija takvim profilom nije moguća, te je analizu potrebno ponoviti. Zaključci: Uzimajući u obzir dug vremenski period od trenutka stradavanja do ekshumacije, DNA analiza je metoda izbora u identifikaciji osoba iz koštanih ostataka. Bez obzira na probleme degradacije i kontaminacije, ona je najpouzdanija, najtočnija, a možda i jedina metoda identifikacije žrtava Drugog svjetskog rata.Aim: To describe in detail the process of identification of skeletal remains from World War II victims using DNA analysis and to point out the problems of DNA isolation from old samples and the problems of finding close relatives for DNA comparison. Methods: DNA from skull and femur samples was isolated using standard phenol/chloroform/isoamyl alcohol extraction. EDTA decalcification procedure was performed before the extraction. DNA quantification was performed on a RT-PCR ABI Prism 7000 Sequence Detection System. The PCR amplification was performed on a GeneAmp® PCR System 9700, using the AmpFLSTR® Identifiler® PCR Amplification Kit. Typing of PCR products was performed on an ABI PRISM ® 310 Genetic Analyzer. Results: Skeletal sample analysis resulted in an incomplete DNA profile. This kind of a DNA profile can not be used for identification and the samples must be reanalysed. Conclusion: Taking into consideration the time period from the moment of death to exhumation, DNA analysis is the method of choice in identification of skeletal remains. Regardless of the problems of degradation and contamination it is the most reliable, the most accurate and maybe the only method of identification of World War II victims