Andelinović Simun, Sutlović Davorka, Erceg Ivkosić Ivana, Skaro Vedrana, Ivkosić Ante, Paić Frane, Rezić Boja, Definis-Gojanović Marija, Primorac Dragan
Split University Hospital, Split, Croatia.
Croat Med J. 2005 Aug;46(4):530-9.
To present twelve-year (1993-2005) experience in identification of human remains found in mass graves in Croatia and Bosnia and Herzegovina (BH), as well as remains that presumably belonged to Croatian citizens given by Serbia and Montenegro. The unique experience of identification of more than a thousand of skeletal samples is valuable for better organization of post-mortem identifications.
Standard forensic methods and methods based on DNA analysis were used for identification of human remains from mass graves. DNA was isolated using standard phenol/chloroform/isoamyl alcohol extraction. In some cases, decalcification and repurification were used prior to the extraction to overcome inhibition of amplification process. Different DNA systems were used for DNA quantitation and amplification (AluQuant, short tandem repeats (STR) commercial systems, Y chromosome STRs, and mitochondrial DNA [mtDNA]). Typing of PCR products was performed on AmpliType PM and AmpliType DQA1 DNA probe strips, ABI PRISM(R) 310 Genetic Analyzer and immobilized sequence-specific oligonucleotide (SSO) probes.
Up-to-date analysis of 1,155 skeletal samples resulted in 703 positively identified bodies: 577 using standard forensic methods, 109 by DNA typing, and 17 by combination of these two methods. The majority of identifications from 1993 to 1999 was, as usual, achieved by standard forensic methods. Later on, these methods were not sufficient and DNA analysis was requested. It was performed in 42% of all cases in 12 years. The crucial step in DNA analysis is extraction of genomic DNA. Standard phenol/chloroform/isoamyl alcohol extraction, complemented with other methods and modifications, proved as the most successful method for this step. In certain cases, the quality and/or quantity of nDNA was not satisfying and the analysis of the mtDNA was performed.
Our experience demonstrated that the advent of forensic DNA analysis methods greatly increased our ability to positively identify previously unknown skeletal remains by a comparative genetic analysis with presumptive relatives.
介绍在克罗地亚和波斯尼亚和黑塞哥维那（波黑）乱葬坑中发现的人类遗骸以及塞尔维亚和黑山移交的据推测属于克罗地亚公民的遗骸的十二年（1993 - 2005年）鉴定经验。对一千多份骨骼样本的独特鉴定经验对于更好地组织死后身份鉴定具有重要价值。
采用标准法医方法和基于DNA分析的方法对乱葬坑中的人类遗骸进行鉴定。使用标准酚/氯仿/异戊醇提取法分离DNA。在某些情况下，提取前进行脱钙和再纯化以克服扩增过程的抑制作用。使用不同的DNA系统进行DNA定量和扩增（AluQuant、短串联重复序列（STR）商业系统、Y染色体STRs和线粒体DNA [mtDNA]）。PCR产物的分型在AmpliType PM和AmpliType DQA1 DNA探针条、ABI PRISM(R) 310遗传分析仪以及固定化序列特异性寡核苷酸（SSO）探针上进行。
对1155份骨骼样本的最新分析得出703具身份已明确的尸体：577具通过标准法医方法鉴定，109具通过DNA分型鉴定，17具通过这两种方法结合鉴定。1993年至1999年的大多数鉴定通常是通过标准法医方法完成的。后来，这些方法已不够用，于是要求进行DNA分析。在12年的所有案例中，42%进行了DNA分析。DNA分析的关键步骤是基因组DNA的提取。标准酚/氯仿/异戊醇提取法，辅以其他方法和改进措施，被证明是这一步最成功的方法。在某些情况下，核DNA的质量和/或数量不令人满意，于是进行了mtDNA分析。
我们的经验表明，法医DNA分析方法的出现极大地提高了我们通过与推定亲属进行比较基因分析来明确鉴定先前未知骨骼遗骸的能力。
