Advanced technique for the preparation of bone and tooth tissue for molecular-genetic expert tests

Laboratory schemes related with the procedure of purifying the osseous and dental samples by applying the dental drilling machine, dental technical carborundum shapes, fogging discs and hard-alloy dental drills were made use of to improve the quality and effectiveness of the mentioned schemes used prior to molecular-and-genetic expertise tests.     

Distribution of HLA DQA1, LDLR,GYPA, HBGG,D7S8, and GC locus alleles in the population of Russia

The distribution of chromosome locus alleles HLA DQA1, LDLR, GYPA, HBGG, D7S8, and GC of PolyMarker molecular genetic individualizing system was studied for the first time in a representative "mean statistical" sampling of Russian population. Typing of these locuses was carried out in 391 donors (no relatives) from 63 regions of the Russian Federation. The incidence of genotypes of all 6 locuses corresponded to the expected values, estimated on the basis of Hardy-Weinberg equilibrium hypothesis. This allows us to use the frequency characteristics of HLA DQA1 locus and the PolyMarker locuses determined in our study as the reference parameters for standard probability estimations in DNA identification. The frequencies of PolyMarker locuses alleles in the Russian sampling (in comparison with other ethnic groups) coincided best of all for allele frequencies in Europeoids living in the USA. For expert evaluation of the efficiency of using these locuses as molecular genetic markers with identification purposes, the discrimination potential was estimated separately for each locus and combinations thereof. HLA DQA1 locus was the most informative of the studied 6 locuses. The main population characteristics of this locus (probability of accidental coincidence, potential of discrimination--PD, polymorphism coefficient--PIC, exclusion potential--Pe, and mean value of parentage index--PI) were estimated for the population of Russia. The frequency distribution of alleles of the studied panel of locuses in the mean statistical Russian population obtained in our study can be used in molecular genetic personality identification and in anthropological studies.     

Utilization of complex technologies of molecular genetics for forensic expert identification of unidentified remains of victims of terrorist acts in Moscow in 1999

This paper presents the first experience gained in Russia in expert identification practice with complex utilization of technologies of molecular genetic individualization of biological objects for solving identification tasks requiring the maximum complete armory of mutually supplementing means. This is particularly important for forensic expert personality identification in cases with many deaths, when indirect identification has to be resorted to, consisting in the use of biological samples from relatives of the victims as the identifying objects. Methodological approaches and concepts of expert studies, used for identification of the remains of people who died in terroristic acts committed in Moscow in September, 1999, are discussed. The results of this study and methodological experience notably extend the potentialities of expert evaluation as regards forensic medical identification of victims of overall disasters, terroristic acts, and war conflicts.     

Research of potentially linked variation of polymorphism of chromosome DNA in aspect of forensic expertise using molecular-genetic individualizing systems CD4, vWA and vWFII

Investigated within the case study are parameters of disbalance of lineage (HC) for 4 micro-satellite locuses of human genome: LPL, CD4, vWA and vWFII. The above locuses are widely used, both in Russia and abroad, in molecular-genetic applications for personality identification. Meanwhile, according to cytogenetics criteria, CD4, vWA and vWFII, are located close to each other in the telomeric region 12pter-12p12 in the short chromosome 12 arm, therefore their potential genetic interdependence is still a topical issue. We found a reliable HC between locuses vWA and vWFII. Locus CD4 did not display HC with locuses vWA and vWFII or with locus LPL. The latter, which is located in chromosome 8 and which must have been negative control for HC, was shown to have no HC with any of the studied markers. Such results correlate well with data on the relative physical localization of CD4, vWA, vWFII and LPL. Multiplication of frequency of alleles (genotypes) is not acceptable in typing locuses vWA and vWFII within one multi-locus panel due to the genetic linkage of these markers demonstrated within the present case study, which is an important practical conclusion.     

Possible unfavourable effects of using in-house components for forensic expert molecular-genetic technologies

So-called in-house production of reagents, components and kits for DNA analysis which are made without strict quality control and standards are now widely practiced in forensic molecular-genetic examinations. Markers of molecular mass were studied to illustrate problems which may arise in use of such in-house components. Other difficulties and negative sequelae of in-house products are also demonstrated.     

Approaches to setting up the Center for identification of exhumed bodies in Chechen Republic

The authors participated in the activity of a group of European experts who visited Moscow, Rostov-on-Don and Grozny in September 2005 to clarify situation with identification of exhumed unknown dead bodies of the civil population. The European experts recommend to set up Center for Identification in Chechen Republic (in Grozny). The authors propose to make DNA identification tests in the Russian Federation Center for Forensic Medical Evaluation in Moscow which has much experience and staff skilled in identification of unknown exhumed bodies and can solve the problem of genetic identification of unidentified bodies of people missed in the Chechen Republic more effectively.     

Development and application of hydrogel oligonucleotide microchips for forensic-medical personal identification as illustrated by ABO locus

The article describes the method defining 5 alleles of ABO blood group typing system by molecular hybridization in hydrogel oligonucleotide microchip. The testing points were SNP variants in positions 261 and 297of exon 6 and in positions 646 and 657 of exon 7. Therefore, 5 ABO blood groups can be easily revealed: A, B, 0(1), 0(1v), 0(2). The method was tested on 10 DNA samples isolated from blood and saliva of unrelated individuals. The results were confirmed by sequencing of the identified allelic fragments. Estimation sensitivity was 25 pg of total DNA input. This technique is cost-effective and easy for use and, therefore, promising for forensic-medical personal identification.