Design of the automatic system of processing data on mitochondrial DNA "mDNAbase" sequestration for expert identification of the unidentified bodies under the conditions of mass reception of human remains

The authors consider utilization of software which provides indirect personality identification by establishment of biological kinship using a comparative analysis of nucleotide sequences (NS) of mitochondrial DNA (mtDNA) in unidentified bodies and suspected relatives of these victims. The software supports data bases with information on NS. To secure automatic search there are modes of cross-processing. Results of identification are formed as lists of exclusions and tables with calculated frequencies of mitotypes of identification objects as well as protocols of a comparative analysis of nucleotide position in mtDNA sequences under comparison. The proposed computer approach is a new highly effective tool for identification of personality based on data on mtDNA analysis in conditions of stream information processing under mass reception of unidentified bodies.     

Mitochondrial DNA typing--a new level for solving identification problems in forensic medical expert identification of unidentified remains of victims of terrorist acts in Moscow and the armed conflict in the Chechen Republic

Two large-scale episodes described in this paper reflect the first in Russia use of molecular genetic matrilinear markers (analysis of polymorphism of sequences of amplified fragments of mitochondrial DNA hypervariable locuses) in solution of a complex identification problem: forensic medical identification of unidentified fragments of victims of explosions of houses in Moscow in September, 1999, and of soldiers dead in the war conflict in the Chechen Republic in 1994-1996. The results of this work and methodological experience gained in it essentially extend the potentialities of expert studies as regards forensic medical identification of victims of large scale disasters, terroristic acts, and war conflicts.     

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.     

Population study of HLA DQA1, LDLR,GYPA, HBGG,D7S8, and GC loci in Caucasians living in the Ural region of Russia

Allele frequencies have been determined for 6 tested locuses. The distribution of HLA DQA1 gene and locuses of Polymarker kit (Perkin Elmer, USA) for population studies conformed to the Hardy-Weinberg equilibrium (HWE). Inequilibrium for linkage for all possible locus pairs was tested by an accurate test. No statistically significant inequilibrium was detected. Paired comparison of allele frequencies of tested locuses of the Ural population with other Caucasoid populations showed homogeneity for GYPA, HBGG, D7S8, and HLA DQA1 locuses. Significant differences for 1 and 2 of the 4 compared pairs were detected for LDLR and GC locuses, respectively. The total discrimination potential for 6 tested locuses was 0.9995. Our population study showed that high informative value of the tested genetic markers and conformity to HWE make them a useful tool for forensic genetic studies in the Ural Caucasoid population. Preliminary assessment of the efficiency of PCR of 6 studied locuses by capillary electrophoresis is highly effective in typing of DNA samples isolated from material evidences with negligible content of biological material with presumably high degree of degradation and for evaluating the positive and negative amplification controls.     

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.     

Computer-aided personality identification by skull and life-time photography by POSKID 1.1 method

An improved method for computer-aided personality identification by the skull, based on the POSKID 1.1 software, consists in investigation of enlarged images of the skull and life-time photograph of the probable individual by coordinates of 49 anatomical points; independent quantitative evaluation of the aspect of each of the compared objects by the X, Y, and Z axes; formal evaluation of the results of comparative study of the skull-portrait by multidimensional discriminant analysis models. The proposed version differs from the POSKID 1.0 software in the method for evaluating the spatial position of the head on the portrait and adequate orientation of the skull in space, which necessitates the utilization of coordinate-regulated holder POSKID 1.1 method is based on multidimensional discriminant analysis and suggests a virtually reliable solution in 76.13-80.65% cases, a probable solution (positive and negative) in 11.61-18.06% cases, and motivated refusal from solution in 5.81-7.74% cases. In case of a probable or indefinite solution further investigations are recommended making use of life-time photographs with different aspects.     

DNA Typing for the Identification of Old Skeletal Remains from Korean War Victims*

Abstract: The identification of missing casualties of the Korean War (1950–1953) has been performed using mitochondrial DNA (mtDNA) profiles, but recent advances in DNA extraction techniques and approaches using smaller amplicons have significantly increased the possibility of obtaining DNA profiles from highly degraded skeletal remains. Therefore, 21 skeletal remains of Korean War victims and 24 samples from biological relatives of the supposed victims were selected based on circumstantial evidence and/or mtDNA‐matching results and were analyzed to confirm the alleged relationship. Cumulative likelihood ratios were obtained from autosomal short tandem repeat, Y‐chromosomal STR, and mtDNA‐genotyping results, and mainly confirmed the alleged relationship with values over 10⁵. The present analysis emphasizes the value of mini‐ and Y‐STR systems as well as an efficient DNA extraction method in DNA testing for the identification of old skeletal remains.     

Genotypic Polymorphisms of Hepatitis B Virus Provide Useful Information for Estimating Geographical Origin or Place of Long‐term Residence of Unidentified Cadavers

Increasing numbers of unidentified cadavers are a major problem. We have developed a new method for providing identification information that can determine the geographical origin or place of long‐term residence of unidentified cadavers based on genotypic polymorphisms of hepatitis B virus (HBV) known to correlate with their geographical distribution. PCR of serum samples detected HBV DNA from 4 (3.9%) of 102 randomly selected Japanese forensic cadavers. Multiplex PCR did not detect multiple HBV genotypes from any single cadaver, confirming the absence of coinfection. Phylogenetic tree analysis based on a 485‐bp mutant region of the HBV S gene successfully classified the HBV genotypes into A to J. Among 10 HBV‐infected cadavers, 8 had genotype Ce/C2, a genotype prevalent in East Asia, and 2 had genotype Bj/B1, a Japanese‐specific genotype. HBV genotypic polymorphisms correlate with the geographical distribution of the virus and thus provide important information for identifying unidentified cadavers infected with HBV.     

Choosing relatives for DNA identification of missing persons

DNA-based analysis is integral to missing person identification cases. When direct references are not available, indirect relative references can be used to identify missing persons by kinship analysis. Generally, more reference relatives render greater accuracy of identification. However, it is costly to type multiple references. Thus, at times, decisions may need to be made on which relatives to type. In this study, pedigrees for 37 common reference scenarios with 13 CODIS STRs were simulated to rank the information content of different combinations of relatives. The results confirm that first-order relatives (parents and fullsibs) are the most preferred relatives to identify missing persons; fullsibs are also informative. Less genetic dependence between references provides a higher on average likelihood ratio. Distant relatives may not be helpful solely by autosomal markers. But lineage-based Y chromosome and mitochondrial DNA markers can increase the likelihood ratio or serve as filters to exclude putative relationships.     

The use of capillary electrophoresis for the study of D13S317 and D7S820 microsatellites in Caucasian population of the Ural region of Russia

A method for the simultaneous amplification and typing of microsatellites D13S17 and D7S820 by using monocomplex sets GenePrint Fluorescent STR Systems ("Promega Corp.", USA) was worked out. The offered method is accurate, reproductive, sensitive and needs a short time for analysis. Allele frequencies D13S17 and D7S820 for the above mentioned Caucasians were determined on the population sample of 120 persons who were not relatives and who resided in the Ural region of Russia. The genotype frequencies of both locuses in this population sample did not differ statistically from Hardy-Weinberg equilibrium. The aggregate discrimination factor for both locuses amounted to 0.995. The null hypothesis of independence among locuses D13S17 and D7S820 was checked by using an accurate test. No relation was found between alleles of the 2 tested locuses in the population sample of Ural Caucasians. The testing of the null hypothesis of the population homogeneity of allele frequencies of the examined locuses showed, among 5 Caucasian population samples, a reliable difference only for locus D7S802 in case of a couple of an Ural and Polish population sample. Allele frequencies for other couples were homogeneous.     

Investigation of Different Ways in Which the CODIS 7.0 May be Used in Mass Disaster Identification

DNA analysis is a key method for the identification of human remains in mass disasters. Reference samples from relatives may be used to identify missing persons by kinship analysis. Different methods of applying the CODIS in disaster victim identification (DVI) were investigated. Two searches were evaluated: (i) relating family relatives to a pedigree tree (FPT) and (ii) relating unidentified human remains to a pedigree tree (UPT). A joint pedigree likelihood ratio (JPLR) and rank were calculated for each search. Both searches were similar in average JPLR and rank. In exceptional cases, namely the existence of a mutation different from the CODIS model, a nonbiological father, a mistake in STR, or incorrect profile association, the UPT search returned one true rank, whereas the FPT search returned no results. This paper suggests a novel strategy to overcome these limitations and increase efficiency in conducting identification of mass disaster victims.     

Testing the reliability of frontal sinuses in positive identification

The use of frontal sinus radiographs in positive identification has become an increasingly applied and accepted technique among forensic anthropologists, radiologists, and pathologists. From an evidentiary standpoint, however, it is important to know whether frontal sinus radiographs are a reliable method for confirming or rejecting an identification, and standardized methods should be applied when making comparisons. The purpose of the following study is to develop an objective, standardized comparison method, and investigate the reliability of that method. Elliptic Fourier analysis (EFA) was used to assess the variation in 808 outlines of frontal sinuses by calculating likelihood ratios and posterior probabilities from EFA coefficients. Results show that using EFA coefficient comparison to estimate the probability of a correct identification is a reliable technique, and EFA comparison of frontal sinus outlines is recommended when it may be necessary to provide quantitative substantiation for a forensic identification based on these structures.     

Enhanced kinship analysis and STR-based DNA typing for human identification in mass fatality incidents: the Swissair flight 111 disaster

A bioinformatic tool was developed to assist with the victim identification initiative that followed the Swissair Flight 111 disaster. Making use of short tandem repeat (STR) DNA typing data generated with AmpFlSTR Profiler Plus (PP) and AmpFlSTR COfiler(CO) kits, the software systematically compared each available STR genotype with every other genotype. The matching algorithm was based on the search for: (i) direct matches to genotypes derived from personal effects; and (ii) potential kinship associations between victims and next-of-kin, as measured by allele sharing at individual loci. The software greatly assisted parentage analysis by enabling kinship evaluation in situations where complete parentage trios were unavailable and, in some situations, with distantly related relatives. Exclusion of fortuitous kinship associations (FKA) was made possible through the recovery at the disaster site of at least one remains for every sought-after victim, and was incorporated into the kinship software. The data from the 13 combined STR loci produced 6 and 23 times fewer FKAs when compared with PP alone and AmpFlSTR Profiler (PR) alone, respectively. Identification leads or confirmations of identification were obtained for 218 victims for which DNA reference samples (personal effects and kin) had been submitted. Confirmation of an inferred kinship association was sought through frequency and likelihood calculations, as well as corroborative data from other identification modalities. The use of a simple, yet powerful, automated genotype comparison approach and the use of megaplexes with high power of discrimination (PD) values extended considerably the identification capabilities in the case of the Swissair disaster. The DNA typing identification modality proved to be a valuable component of the large arsenal of identification tools deployed in the aftermath of this disaster.     

Genomic "dactyloscopy" in the expertise of disputed paternity and the determination of biological relationship

Genome "dactyloscopy" (DNA fingerprinting) is a principally new way of personal identification based on analysis of human genetic material (DNA); the difference in DNA structure of different subjects is the scientific basis of this method. This ensures opportunity to estimate biological relationship of persons positively. The authors were the first to demonstrate using certain expert material the adequacy and potentials of DNA fingerprinting by M 13 probe for medicolegal expert practice in most complicated cases of relationship determination requiring positive identification of paternity and maternity.     

Studies of the allelic diversity in micro-satellite locus D16S539, F13B, FESFPS, TH01, and TROX in European population of Russia Ural region using capillary electrophoresis

Allelic frequencies of chromosome micro-satellite locuses D16S539, F13B, FESFPS, TH01 and TPOX were determined, within the case study, in a sampling of Europeoidal individuals residing in Russia's Ural Region. The allelic variants were analyzed by capillary electrophoresis after the enzyme amplification in polymerase chain reaction with fluorescent primers. The genotypic frequencies of the studied locuses were shown not to divert with statistical reliability from Hardy-Weinberg equilibrium. The estimated aggregate discriminative potential for a panel of 5 studied locuses made 0.99995. No nonequilibrium was found by linkage between alleles of all lucuses examined within the present case study or between their alleles and the alleles of previously investigated locuses D7S820 and D13S317. The implemented testing of the population homogeneity of allelic frequencies of investigated locuses for 3 samplings of Europeoids showed a deviation for locus FESFPS versus the Ural and Polish samplings and for locus F13B in the Ural and North America samplings. The distribution of allelic frequencies of other locuses was homogenous in the compared samplings.     

An algorithm of computerized processing of molecular-genetic data for personality identification based on the outcome of events involving numerous victims

Computerized systems of data processing are a high-efficiency tool for the storage and processing of big volumes of molecular genetic data as well as for the forensic medical identification of personality in events involving enormous unidentified bodies coming from zones of calamity or natural disaster. Described in the paper is a sequence of actions undertaken by the expert when the DNAdacto and mDNAbase soft tools are in use. The algorithm of computer-based expert analysis of typing of chromosome and mitochondrial DNA was tested at the 124th Central laboratory of medical-criminal identification of Russia's defense ministry; it was proven to be highly effective for identification made in events with numerous unidentified bodies.     

似然比在法庭说话人确认中的应用

本文把成功应用于DNA检验的证据评估方法（似然比）应用于法庭语音证据评估之中,提取语音的LPC作为识别特征,并使用45人电话对话录音中元音/a/作为样本进行了测试。结果表明该方法不仅能正确识别说话人,而且能根据当前嫌疑人样本和问题语音样本的差异,量化该语音样本作为证据的力度,为法庭提供科学合理的证据评估结果和科学解释。同时,自动特征提取的引入比起人工提取共振峰特征,提高了工作效率,识别系统性能也获得大幅提升。     

The Probabilistic Genotyping Software STRmix: Utility and Evidence for its Validity

Forensic DNA interpretation is transitioning from manual interpretation based usually on binary decision‐making toward computer‐based systems that model the probability of the profile given different explanations for it, termed probabilistic genotyping (PG). Decision‐making by laboratories to implement probability‐based interpretation should be based on scientific principles for validity and information that supports its utility, such as criteria to support admissibility. The principles behind STRmix? are outlined in this study and include standard mathematics and modeling of peak heights and variability in those heights. All PG methods generate a likelihood ratio (LR) and require the formulation of propositions. Principles underpinning formulations of propositions include the identification of reasonably assumed contributors. Substantial data have been produced that support precision, error rate, and reliability of PG, and in particular, STRmix?. A current issue is access to the code and quality processes used while coding. There are substantial data that describe the performance, strengths, and limitations of STRmix?, one of the available PG software.     

A new algorithm for the establishment of blood relationship based on the application of the dermatoglyphic technique

An original classification of dermatoglyphic features is proposed taking into consideration their inter-relations in conjunction with the new principles of comparative studies. A new and improved algorithm for the establishment of blood relationship has been developed based on the application of the dermatoglyphic technique. The method is recommended for the use in forensic medical expertise for the purpose of estimation of the probability of paternity, maternity, and substitution of children. Moreover, it may find application for the indirect (via establishment of blood relationship) identification of personality of an unknown subject.     

On identification problems requiring linked autosomal markers

This paper considers identification problems based on DNA marker data. The topics we discuss are general, but we will exemplify them in a simple context. There is DNA available from two persons. There is uncertainty about the relationship between the two individuals and a number of hypotheses describing the possible relationship is available. The task is to determine the most likely pedigree. This problem is fairly standard. However, there are some problems that cannot be solved using DNA from independently segregating loci. For example, the likelihoods for (i) grandparent–grandchild, (ii) uncle–niece and (iii) half-sibs coincide for such DNA data and so these relations cannot be distinguished on the basis of markers normally used for forensic identification problems: the likelihood ratio comparing any pair of hypotheses will be unity.Sometimes, but not in the examples we consider, other sources of DNA like mtDNA or sex chromosomes can help to distinguish between such equally likely possibilities. Prior information can likewise be of use. For instance, age information can exclude alternative (i) above and also indicate that alternative (iii) is apriori more likely than alternative (ii).More generally, the above problems can be solved using linked autosomal markers. To study the problem in detail and understand how linkage works in this regard, we derive an explicit formula for a pair of linked markers. The formula extends to independent pairs of linked markers. While this approach adds to the understanding of the problem, more markers are required to obtain satisfactory results and then the Lander–Green algorithm is needed. Simulation experiments are presented based on a range of scenarios and we conclude that useful results can be obtained using available freeware (MERLIN and R).The main message of this paper is that linked autosomal markers deserve greater attention in forensic genetics and that the required laboratory and statistical analyses can be performed based on existing technology and freeware.