Turkiye Klinikleri Journal of Forensic Medicine and Forensic Sciences

.: ORIGINAL RESEARCH
Karışım DNA Örneklerinde STR Analizi ve Yorumlanması
STR Analysis and Interpretation of Dna Mixture Samples
Tolga ZORLUa, Özlem BÜLBÜLa, Gönül FİLOĞLUa
aİstanbul Üniversitesi-Cerrahpaşa Adli Tıp Enstitüsü, İstanbul, TÜRKİYE
Turkiye Klinikleri J Foren Sci Leg Med. 2019;16(2):104-19
doi: 10.5336/forensic.2019-64962
Article Language: TR
Full Text
ÖZET
 Amaç: Adli vakaların değerlendirilme sürecinde birden fazla kişinin DNA'sını içeren karışım örnekleri ile sıklıkla karşılaşılmaktadır. İki veya daha fazla kişinin genetik profilinin bir arada bulunduğu bu tür örneklerde, bazı faktörler (katılımcı sayısının bilinmemesi, allel çakışmalarının olması, düşük kalitede ve bozunmuş örnekler vb.) DNA analizini daha karmaşık hâle getirebilmektedir. Son yıllarda çeşitli ülkelerdeki hukuk sistemleri karışım DNA örneklerinin yorumlanmasında istatistiksel hesaplama işlemlerinden faydalanmaktadırlar. Ülkemizde karışım DNA analizlerinde kullanılacak herhangi bir prosedür bulunmamaktadır. Bu çalışmada; karışım örneklerinde profil olasılığı (GeneMapper® IDX v.1 Mixture Analysis) ve olabilirlik oranı (Forensim LRmix?) kullanılarak, hem doğru genotipleme yapmak hem de katılımcı sayısını doğru bir şekilde belirlemek amaçlanmıştır. Gereç ve Yöntemler: Bu çalışmada, gönüllü kişilerden toplanan kan örneklerinden değişik oranlarda (1:1, 1:3, 1:5, 1:10, 1:20, 1:50) karışım DNA örnekleri oluşturularak valide edilen AmpFℓSTR® Identifiler kiti ile çoğaltıldı. Polimeraz zincir reaksiyonu (PCR) ürünleri AB 3130 genetik analizörde yürütüldü ve Genemapper® IDX v.1 programında analiz edildi. Karışım örneklerini oluşturan katılımcıların tiplendirilmesi ve karışım oranlarının tahmin edilmesi Genemapper ® IDX v.1. Mixture Analysis ile yapıldı. Karışım örneklerinin allel düşmesi aralığına göre performans testi ve katılımcıların örnek profilinde bulunma olasılıklarının istatistiksel değerlendirmesi Forensim LRmix? uygulaması ile yapıldı. Bulgular: Analizde en sık karşılaşılan problem allel düşmesi olarak saptanmıştır. Olabilirlik oranı yaklaşımı ve Forensim LRmix? uygulaması kullanılarak oluşturulan karışım örneklerinin doğru bir şekilde DNA analizinin yapılabildiği belirlenmiştir. Sonuç: Karışım DNA örneklerinin objektif bir şekilde yorumlanabilmesi ve delilin gücünü ortaya koymak için istatistiksel yöntemler kullanılmalıdır. Bu çalışma; karışım DNA örneklerinin gelişmiş dünya ülkelerinde olduğu gibi ülkemizde de benzer standartlarda yorumlanabilmesi için yapılan öncü bir araştırmadır.

Anahtar Kelimeler: Adli genetik; kompleks DNA karışımları; mikrosatelit tekrarları; STR profili; LRmix modulu
ABSTRACT
 Objective: In the process of evaluation of forensic cases, samples of the mixture containing more than one person are frequently encountered. In such examples of mixtures where the genetic profile of two or more persons are combined, some factors (Unknow number of contributors, presence of same alleles, poor quality and degraded samples, etc.) may further complicate DNA analysis. In recent years, legal systems in various countries have benefited from statistical calculations in the interpretation of mixture DNA samples. In our country, there are no procedures to be used in mixture DNA analysis. The aim of this study; to determine the correct genotypes and the number of contributors using the profile probability (GeneMapperR IDX v.1 Mixture Analysis) and likelihood ratio (Forensim LRmix?) in the mixture samples. Material and Methods: In this study, mixture samples were evaluated in different ratios (1:1, 1:3, 1:5, 1:10, 1:20, 1:50) from the blood samples which collected from volunteers. The polymerase chain reaction (PCR) was applied using validated AmpFℓSTRR Identifiler kit. PCR products were run in the AB 3130 enetic analyzer and analyzed in the GenemapperR IDX v.1 program. Typing of the contributors and estimating the mixture ratios were done on GenemapperR IDX v.1. Mixture Analysis. Forensim LRmix? application was used to test the performance of the mixture samples according to the allele drop interval and to evaluate the probabilities of the participants in the sample profile. Results: The most common problem in the analysis was the allele drop-out. Likelihood ratio and Forensim LRmix module was determined as a useful and successful application on both mixture samples that composed on controlled conditions. Conclusion: As a result, statistical methods should be used to interpret mixture DNA samples and to demonstrate the strength of evidence. This work as a pioneering research conducted in order to interpret mixed DNA samples in similar standards in our country as in developed countries.

Keywords: Forensic genetic; complex DNA mixtures; microsatellite repeats; Short tandem repeats; LRmix module
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