[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). Markers were selected to meet the following criteria: the minor allele frequency (MAF) is higher than 0.30; the physical distance between markers is greater than 3 Mb; there are no polymorphisms, tandem repeats, and palindromes in the flanking sequences; the AT/GC ratio is close to 1. A panel of 106 polymorphisms was thus formed; the average MAF was estimated at 0.396 in the five populations. The method developed for panel genotyping included one-step multiplex PCR and subsequent hybridization on a biological microarray. The average amplicon length was 72 bp. A sample of 201 residents of Moscow and St. Petersburg was tested to determine the main characteristics of the panel: the random matching probability (MP) was 1.89x 10^(-43) and the combined probability of paternity exclusion (CPE) was 0.99999999063. The method provides an alternative to molecular genetic personal identification based on the STR length variations.

[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 setting protocol is simplified as much as possible to facilitate the introduction of the method into practice. The distribution of allele frequencies of the studied polymorphisms, as well as AB0 blood groups among the Slavs (N = 482), originating mainly from central Russia, was established.

[The molecular-genetic analysis of mitochondrial DNA from the burnt bones: 'the limits of the possible' problem revisited]. / Molekuliarno-geneticheskii analiz mitokhondrial'noi DNK v obozhzhennykh kostiakh: eshche raz o predelakh vozmozhnogo.

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. In the cases of a more rigorous thermal impact when the bone tissue exhibits well pronounced signs of heat destruction, it should be considered as inherently unsuitable for genotyping of mtDNA. It was shown that chromosomal DNA is inferior to mtDNA in terms of heat resistance. This finding agrees with the currently adopted view, however this advantage of mtDNA is relatively insignificant from the standpoint of genotyping efficiency.

[On the problem of HIV infection in the objects of forensic medical expertise]. / O probleme diagnostiki VICh-infektsii v ob''ektakh sudebno-meditsinskoi ékspertizy.

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 special attention is given to the advantages of the molecular genetic methods based on the use of the specific fragments of the viral RNA genome as the diagnostic markers. The solid methodological basis for molecular genetic diagnostics of HIV infection is provided by PCR-amplification with the detection in the real-time regime. It is supposed that this approach will make it possible not only to determine, with the high degree of accuracy and specificity, the presence of the viral genome in the biological materials but also to reduce to a minimum the probability of both false-positive and false-negative responses.

[The molecular genetic analysis of chromosomal DNA in burned bones]. / Molekulyarno-geneticheskii analiz khromosomnoi DNK v obozhzhennykh kostyakh: mif ili real'nost'?

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. in the carbonized, grey- and white-burnt states) were placed in the muffle furnace for the controlled thermal treatment. The analytical suitability of these burned bone objects for genotyping was estimated with the use of standard chromosomal STR-loci multiplex genotyping panels. The results of the study cast serious doubts as regards the possibility of genotyping of chromosomal DNA extracted from the burned bones. It was shown that the exposure of the bone tissue to a temperature of 150 degrees Celsius during 2 hours can turn it into a material absolutely unsuitable for genotyping due to the loss of all individual genotypic traits. Characteristically the burned bone objects are externally indistinguishable from the native bone. At the same time, the material with the signs of the high-temperature impact visible by the unaided eye (e.g. in the carbonized, pronounced black as well as grey and white-burnt states) is altogether unsuitable for the reliable identification of the genetic profile of chromosomal DNA.

[The interpopulation differences between nucleotide sequence polymorphisms in alleles of the STR-loci of human chromosomal DNA]. / Mezhpopulyatsionnye razlichiya polimorfizma nukleotidnoi posledovatel'nosti v allelyakh STR-lokusov khromosomnoi DNK cheloveka.

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. Some of the loci of interest were found to have allelic variants the occurrence of which was significantly different in individual samples. Such alleles are of importance for the further investigation since they can be regarded as potential ethno-geographical markers. Their application opens up the new promising prospects for the expert detection of the ethnic affiliation of individual subjects.

[Polymorphism of DNA nucleotide sequence as a source of enhancement of the discrimination potential of the STR-markers]. / Polimorfizm nukleotidnoi posledovatel'nosti DNK kak istochnik uvelicheniya diskriminiruyushchego potentsiala STR-markerov.

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 study has demonstrated that polymorphism of DNA nucleotide sequence can be regarded as the possible source of enhancement of the discriminating potential of STR markers. It means that the analysis of polymorphism of DNA nucleotide sequence for genotyping AFLP-type markers of chromosomal DNA can considerably increase the effectiveness of their application as individualizing markers for the purpose of molecular genetic expertises.

[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 reference objects were the contact traces left on the personal belongings by their owner. The aggregate probabilistic estimate of the coincidence of genotype traits of autosomal DNA, Y-chromosomal DNA, and mtDNA was at least 99,(9)29 4% which gives evidence of the genetic identity of the objects of study. It is this value (99.999999 <...> 9999999(29) 4%) that characterizes the probability that the bone fragments provided for the laboratory studies are actually authentic remains of Yasser Arafat.