Dragana Zgonjanin

Sequence polymorphism of the mitochondrial DNA control region in the population of Vojvodina Province, Serbia

In order to generate and establish the database for forensic identification purposes in Vojvodina Province (Serbia), the sequence of the hypervariable regions 1 (HV1) and 2 (HV2) of the mtDNA control region were determined in a population of 104 unrelated individuals from Vojvodina Province, using a fluorescent-based capillary electrophoresis sequencing method. A total of 93 different haplotypes were found, of these 83 mtDNA types were unique, nine haplotypes were shared by two individuals and one haplotype by three individuals. The variation of mtDNA HV1 and HV2 regions was confined to 116 nucleotide positions, of which 72 were observed in the HV1 and 44 in the HV2. A statistical estimate of the results for this population showed the genetic diversity of 0.9977 and the random match probability of 1.18%. Haplogroup H was the most common haplogroup (43.3%). Haplogroups observed at intermediate levels included clusters U (13.5%), T (10.6%), J (8.6%) and W (5.8%).

Ethanol Concentrations in Antemortem Blood Samples Under Controlled Conditions

AIM The change of antemortem blood alcohol concentration (BAC) in inadequately processed samples was examined. METHODS The study was performed on nine healthy, sober volunteers after overnight fasting. Blood samples were divided into groups and stored, with or without NaF, for varying time periods (12, 24 and 48 h) and at different temperatures (4 degrees C and 20 degrees C). BAC analysis was performed by the gas-chromatography method. RESULT All groups showed very low values of BAC. CONCLUSION Surprisingly, this study showed no ethanol production in improperly processed antemortem blood samples in healthy and sober individuals who were subjected to overnight fasting.

Genetic characterization of 27 Y-STR loci with the Yfiler ® Plus kit in the population of Serbia

Recently, a new 6-dye multiplex, AmpFlSTR Yfiler Plus PCR Amplification Kit (Yfiler Plus, Applied Biosystems, USA) has been developed, which can co-amplify 27 Y-STR loci. 17 Y-STRs widely used loci included in the AmpFlSTR Yfiler Amplification Kit (Yfiler, Applied Biosystems, USA) are included in this kit, besides 10 new additional Y-STR loci (DYS481, DYS460, DYS533, DYS449, DYS576, DYS627, DYS518, DYS570, and DYF387S1a/b), with the last seven being rapidly mutating Y-STRs (RM Y-STRs) [1]. These RM YSTRs are very useful for the discrimination between closely related and unrelated males [1–4]. In this study, we used the Yfiler Plus kit to type 203 unrelated individuals from 3 geographically different areas of Serbia; Northern Serbia, Central Serbia and Southern Serbia (Fig. S1). DNA was obtained from bloodstains or buccal swabs by using the Chelex 100 extraction method [5] from healthy unrelated individuals; 68 from Northern Serbia, 68 from Central Serbia and 67 from Southern Serbia, born and residing in the selected geographical areas, following informed consent. PCR amplification was performed using a GeneAmp PCR System 9700 Gold Plate (Applied Biosystems, USA) using Yfiler Plus kit, according to manufacturer’s recommendations with modifications in the reaction volume (12.5 ml). The electrophoresis was carried out on an Applied Biosystems 3500 Genetic Analyzer (Applied Biosystems, USA) and the fragment analysis was performed with GeneMapper IDX v.1.4 software (Applied Biosystems, USA). Allele designations were made using the allelic ladders provided with the Yfiler Plus kit, and following the recommendations of the DNA Commission of the ISFG on Y-STR analysis [6,7]. Following the guidelines of Forensic Science International Genetics, data were submitted to the YHRD (Y-chromosomal haplotype reference database; https://yhrd.org [8]) and received the accession number YA004149. The contributor successfully passed the YHRD quality control test. The Y-STR haplotypes observed in the three Serbian geographic areas is reported in Table S1. A total of 200 different haplotypes was observed in the sample of 203 Serbian individuals. A total of 197 unique haplotypes (singletons) were obtained from 203 individuals using the Yfiler Plus kit (Table S2). All haplotypes in Northern, Central and Southern Serbia were unique, but three haplotypes shared between different Serbian regions were found (Table S1). A null allele was observed in one individual at DYS391. In all the samples, four different intermediate alleles were found in two loci (DYS390 and DYS458) whereas locus DYS458 showed the highest number (three different intermediate alleles with