[Universal Panel of Insertion/Deletion Polymorphisms and Biochip-Based Kit ChipID106 for Genetic Personal Identification].

A panel of 106 insertion/deletion (InDel) polymorphisms and a method of their genotyping on biochips were proposed as a new approach to genetic personal identification. Short lengths and low mutation rates are basic properties of InDel markers, which thus have significant advantages over short tandem repeats (STRs) widely used in forensics. The allele frequency distributions of all known InDel polymorphisms were studied in the five largest world populations (European, East Asian, South Asian, African, and American).

[A Biochip for Genotyping Polymorphisms Associated with Eye, Hair, Skin Color, AB0 Blood Group, Sex, Y Chromosome Core Haplogroup, and Its Application to Study the Slavic Population].

This paper presents a method for genotyping a panel of 60 single nucleotide polymorphisms (SNPs) using single-stage PCR followed by hybridization on a hydrogel biochip. The pool of analyzed polymorphisms consists of 41 SNPs included in the HIrisPlex-S panel, 4 SNPs of the AB0 gene (261G>Del, 297A>G, 657C>T, 681G>A), markers of the AMELX and AMELY genes, and 14 SNP markers of the Y chromosome haplogroups: B (M60), C (M130), D (CTS3946), E (M5388), G (P257), H (M2920), I (U179), J (M304), L (M185), N (M231), O (M175), Q (M1105), R (P224) and T (M272). These genetic data allow one to predict the phenotype of the desired person according to the characteristics of eye, hair, skin color, AB0 blood group, sex, and genogeographic origin in the male line.

[The molecular-genetic analysis of mitochondrial DNA from the burnt bones: 'the limits of the possible' problem revisited].

The authors report the results of the demonstrative study continuing the cycle of interactive discussions pertinent to the possibility of obtaining reliable genetic information from the analysis of burnt bone fragments. Special emphasis is placed on the worthiness of these materials for genotyping of mitochondrial DNA (mtDNA) with the use of the standard analytical methods employed for the purpose of forensic medical expertise to investigate into the length polymorphism of the amplified mtDNA fragments (PAF) by means of sequencing with fluorescent detection. The study has demonstrated that the mtDNA fragments in the state suitable for genotyping can be found only in the preparations from the bone tissue exposed to the 'mild' thermal impact after which the affected bone is virtually indistinguishable from the native one as far as the outward appearance is concerned.

[On the problem of HIV infection in the objects of forensic medical expertise].

The authors overview the current state of research in the field of diagnostics and identification of the signs suggesting the presence of HIV in the materials obtained from the human corpses undergoing forensic medical expertise at different stages of their post-mortem changes. Another objective of the present work was to evaluate the risk of HIV infection for the medical personnel involved in the autopsy studies taking into consideration the significance attached in different countries to the problem of anti-infectious protection of the staff of the state institutions of forensic medical expertise. The authors discuss the possibilities and limitations of the application of the methods for HIV diagnostics, such as immunoenzymatic assays.

[The molecular genetic analysis of chromosomal DNA in burned bones].

The objective of the present study was a demonstrative consideration of the debatable problem concerning the possibility of obtaining reliable genetic information by the investigation of burned bones. The bone fragments with the identifiable external features of different degree of ignition (i.e.

[The interpopulation differences between nucleotide sequence polymorphisms in alleles of the STR-loci of human chromosomal DNA].

The objective of the present work was to study the phenomenon of nucleotide sequence polymorphism in alleles of the STR-loci of human chromosomal DNA and to estimate its interpopulation differences with a view to the search for the molecular-genetic markers to be used as an efficient tool for the determination of belonging of the subjects of interest to a given population. We undertook the comprehensive analysis of amplified DNA fragment sequence polymorphism (AFSP) and amplified DNA fragment length polymorphism (AFLP) with the use of the PLEX-ID-TM analytical mass-spectrometry platform (Abbott Molecular, USA). The interpopulation differences were estimated in terms of the presence or the absence of single nucleotide replacements (SNP) in the STR markers based on a few population samples.

[Polymorphism of DNA nucleotide sequence as a source of enhancement of the discrimination potential of the STR-markers].

The objective of the present pilot investigation was to reveal and to study polymorphism of nucleotide sequence in the alleles of STR loci of human autosomal DNA with special reference to the role of this phenomenon as a source of the differences between homonymous allelic variants. The secondary objection was to evaluate the possibility of using the data thus obtained for the enhancement of the informative value of the forensic medical genotyping of STR loci by means of identification of single nucleotide polymorphisms (SNP) for the purpose of extending their allelic spectrum. The methodological basis of the study was constituted by the comprehensive amplified fragment length polymorphism (AFLP) analysis and amplified fragment sequence polymorphisms (AFSP) analysis of DNA with the use of the PLEX-ID^TM analytical mass-spectrometry platform (Abbot Molecular, USA).

[The death of Yasser Arafat: molecular-genetic authentication of the remains as an indispensable condition for the evaluation of the medical hypotheses of the cause of his death].

The objective of the present study was the molecular-genetic authentication of the remains as an indispensable condition for the evaluation of the medical hypotheses of the cause of death in 2004 of Yasser Arafat, the former Palestinian leader and the first president of the Palestinian National Administration, the Nobel Peace Prize laureate. We carried out molecular-genetic investigations aimed at establishing the circumstances and cause of the death of Yasser Arafat including the analysis of the relevant medical documentation, the examination of the burial place at Ramallah, remains, and personal belongings stored in his Al Muqata'ah residence at Ramallah. The objective of the present molecular- genetic investigations was to confirm the authenticity of the fragments of Yasser Arafat's remains available for radio-toxicological, chemical toxicological, and other laboratory studies.

[The evaluation of the prospects for the application of mass-spectrometric analysis of the amplified DNA fragments for the purpose of forensic medical expertise].

The objective of the present study was to evaluate the prospects for the application of the mass-spectrometric analysis for the solution of the problems facing modern forensic-medical genetics as illustrated by the example of the new experimental multiplex approach to the typing of human DNA with the use of the complex PLEX-ID platform. The validation study involved all stages of the processing chain. The results of the study were used to develop the recommendations for the optimization of the analytical system being used.

[Multilocus genotyping of polymorphous STR-loci of chromosomal DNA in individual cells: technical difficulties].

This study was designed to estimate the effectiveness of special technical procedures for the enhancement of sensitivity of multiplex analysis of DNA, such as the use of low-plexity PCR systems and the whole genome preamplification technology, and the possibility of their application for the purpose of forensic medical genotyping of polymorphous STR-loci of chromosomal DNA in individual cells. The authors refused to use the imitation model (equivalent DNA dilutions) for the sake of obtaining the maximally informative data and chose to work with real preparations of solitary buccal epithelial cells isolated by the laser microdissection technique. It was shown that neither the use of the low-plexity multilocus PCR systems nor the whole genome pre-amplification technology makes possible reliable genotyping of STR-loci of chromosomal DNA in individual cells.

[Certain topical aspects of the practical application of the laser microdissection technology for the purpose of forensic molecular-genetic analysis].

Experiments with the use of the laser capture microdissection (LCM) technology for the purpose of forensic molecular-genetic analysis carried out on real objects revealed a number of specific aspects of practical LCM application. Some of these problems have been investigated in the present work with special reference to the characteristics of the cells of interest and their physical properties in study objects. The data obtained give reason to conclude that a failure of genotyping of individual cells obtained by the LCM technique may be due to the lack of genetic material suitable for analysis.

[The estimation of possibilities for the application of the laser capture microdissection technology for the molecular-genetic expert analysis (genotyping) of human chromosomal DNA].

The present study was designed to estimate the possibilities of application of the laser capture microdissection (LCM) technology for the molecular-genetic expert analysis (genotyping) of human chromosomal DNA. The experimental method employed for the purpose was the multiplex multilocus analysis of autosomal DNA polymorphism in the preparations of buccal epitheliocytes obtained by LCM. The key principles of the study were the application of physical methods for contrast enhancement of the micropreparations (such as phase-contrast microscopy and dark-field microscopy) and PCR-compatible cell lysis.

[Typing of mitochondrial DNA from isolated cells of the human buccal epithelium].

The authors have developed a method for molecular-genetic analysis of DNA from isolated cells for the purpose of forensic medical diagnostics. The method is based on the use of the laser capture microdissection (LCM) technology in combination with typing of mitochondrial DNA. Optimization of the conditions for amplification of polymorphic mtDNA loci in preparations containing minimal amounts of the genetic material was accomplished at the initial stage of the work.

[Optimization of the biological microchip for genotyping the AB0 locus: analytical aspects].

The present work continues the search for methodological options facilitating the improvement and optimization of the biological microchip designed for genotyping the AB0 locus. It was shown in an earlier study designed to test a prototype biological microchip using a reference set of preparations with the known group specificity that under certain conditions some cells of the biochip appear to generate artifact hybridization signals that tend to make the results of genotyping either incorrect or difficult to interpret. We performed the correction of the molecular structure of DNA probes of the prototype biochip for the purpose of optimization of their hybridization potency.

[Optimization of the biological microchip for genotyping of the AB0 locus: correction of DNA probes].

The objective of the present work was to search for methodological options facilitating the improvement and optimization of the biological microchip designed for genotyping of the AB0 locus. Testing a prototype biological microchip for genotyping of the AB0 locus using a reference set of preparations with the known group specificity has demonstrated that the choice of DNA probes by theoretical calculation of their thermodynamic parameters does not necessarily yields the desired practical result. Suffice it to say that under certain conditions some cells of the biochip appear to generate artifact hybridization signals that tend to make the results of genotyping either incorrect or difficult to interpret.

[Some practical notes about performing polymerase chain reaction and electrophoretic fractionation of amplificated products].

Practical aspects concerning standardization of molecular-genetic expertise performing with the use of the method of DNA are considered. Examples of difficulties, which can occur at nonobservance of requirements of polymerase chain reaction and electrophoresis performing, are described; practical recommendations of their elimination are given.

[Typing of ABO locus with biological microchip as a new level in solution of problems in forensic-medical biological expertise of material evidences].

There are cases in practice when during expertise of material evidences, discrepancies between results of typing of ABO antigens and molecular-genetic typing of DNA occur. In this work, as a radical approach to objective solution of similar conflict situations, for some contradictory case of expertise, all examinations were performed on the unified methodological base--DNA level. Instead of biological (isoserological) typing of ABO antigen, molecular-genetic typing of ABO locus with biological microchip was performed.

[Combined use of molecular genetic technologies to identify the Russian citizens died during tsunami in Thailand].

Main stages, methodological aspects and practical results of molecular genetic research to identify the citizens of Russia died during tsunami in Thailand are considered.

[Properties of the molecular-genetic individual identification system on the basis of D1S111 locus in terms of forensic-expert typing of DNA].

The article presents a comparative analysis of reference nucleotide sequences for locus D1S111, estimation of basic parameters of this locus polymorphism in the representative sample of Russian population for use as an individual identification molecular-genetic system in forensic expert examinations.

[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.

[A definition of the nomenclature of allele variants and a description of allele polymorphism within a molecular-genetic individualization system based on the pentanucleotide tandem replication of HUMCD4].

While analyzing the available published data, we found significant differences in definition of alleles of the HUMCD4 polymorphic chromosome locus. It is an obstacle for comparing the expertise results obtained while using the locus as molecular individualization system in different laboratories and, as a consequence, it hinders the use of the said marker in building up a reference database. The structure of the HUMCD4 locus was analytically investigated and the distribution of the locus alleles was systemized in a sample of 407 persons (citizens of Russia who are not blood relatives) within the present case study for the purpose of a detailed definition of its allele's characteristics.

[Results of applying molecular-and-genetic markers of chromosomal DNA in identification of unrecognizable remains of servicemen lost in military actions in the Northern Caucasus].

The results of applying molecular-and-genetic markers of chromosomal DNA in identifying unrecognizable remains of servicemen lost as dead in 1994-1996 and 1999-2001 military campaigns in the Northern Caucuses are summarized in the paper. Some of the specific features related with enzymatic amplification typing of DNA preparations sampled from degraded biological tissues of strongly deformed or decayed cadavers were analyzed. The typing results were analyzed by the AB0 system of degraded expertise samples in order to check the reliability of routine forensic-biological examinations as applicable to cadaver tissues with pronounced putrefactive changes.

[Study of species specificity of the amelogenin system for genetic sex determination].

The properties of amelogenin amplification system and, in particular, of its species specificity, were studied. DNA preparations extracted from cattle (cow/bull), pig, ram and from poultry (hen), as well as from dog and cat, were used as a matrix for polymerase chain reaction (PCR) involving a standard scheme of enzymatic amplification of the amelogenin gene. It was demonstrated that, unlike for the human DNA, the amelogenin test couldn't be used for the DNA of examined animals as a sex-specific marker.

[Analysis of genetic diversity of the mitochondrial DNA in the forensic medical expert identification of individuals].

Spectra of haplotype frequencies were studied for locuses of hypervariable segments 1 and 2 (HVS1 and HVS2), separately for each, and for linked segment HVS1-HVS2. The obtained data were used to determine the values and to evaluate comparatively the discriminating characteristics of the corresponding individualizing systems based on the typing of mtDNA. The system of typing, based on HVS2 (mv = 0.

[Distribution of D1S80 alleles in a random sample of the population of the Russian Federation].

The distribution of alleles of chromosomal locus D1S80 was studied within a random selection of population of the Russian Federation. Typing was made for 255 persons who were not blood relatives and who lived in 56 Russia's regions. A correlation was found between the observed frequencies of genotypes and those theoretically expected and estimated on the basis of the hypothesis of Hardy-Weinberg equilibrium.