Identification of victims of the 1998 Taoyuan Airbus crash accident using DNA analysis

In February 1998 a civilian aeroplane carrying 196 individuals crashed in Taiwan and killed another 6 people on the ground. Although there were dental and medical records, fingerprints, photographic evidence and personal effects to identify some of the victims, DNA analysis was required to further identify severely damaged remains. From the 202 people known to have perished in the plane crash, a total of 685 fragments of human remains were subjected to DNA analysis. The analysis was carried out using nine microsatellite loci, plus amelogenin to cluster the 685 fragments into 202 groups, accounting for all the victims. To establish genetic relatedness of the victims to other victims and living relatives, additional DNA loci were used. In this case the paternity index was increased by using HLA DQA1 plus Polymarker. The same 16 DNA loci were used to test blood samples from 201 relatives to establish parent/child and sibling relationships. With the exception of 19 victims identified by non-genetic evidence, 183 victims were successfully identified by DNA typing with relatively high values of paternity index by the direct or indirect comparison of relatives. The 202 victims were from 37 different families, ranging in size from 2 to 13 members and 74 individuals known to be unrelated to any other victim. The DNA from living relatives was used to identify one member of a family group, from which other victims of the family could be identified. ABO blood group information was further used to confirm genetic relatedness within families. A comparison of the DNA profiling results to the ABO blood group of the victims showed no discrepancies with the exception of two mutations in the FGA locus. In cases of severely damaged victims from a plane crash, DNA analysis proved to be the best choice to identify victims. Received: 19 August 1998 / Received in revised form: 4 January 1999 / Accepted: 10 March 1999     

Molecular genetic identification of skeletal remains from the Second World War Konfin I mass grave in Slovenia

This paper describes molecular genetic identification of one third of the skeletal remains of 88 victims of postwar (June 1945) killings found in the Konfin I mass grave in Slovenia. Living relatives were traced for 36 victims. We analyzed 84 right femurs and compared their genetic profiles to the genetic material of living relatives. We cleaned the bones, removed surface contamination, and ground the bones into powder. Prior to DNA isolation using Biorobot EZ1 (Qiagen), the powder was decalcified. The nuclear DNA of the samples was quantified using the real-time polymerase chain reaction method. We extracted 0.8 to 100 ng DNA/g of bone powder from 82 bones. Autosomal genetic profiles and Y-chromosome haplotypes were obtained from 98% of the bones, and mitochondrial DNA (mtDNA) haplotypes from 95% of the bones for the HVI region and from 98% of the bones for the HVII region. Genetic profiles of the nuclear and mtDNA were determined for reference persons. For traceability in the event of contamination, we created an elimination database including genetic profiles of the nuclear and mtDNA of all persons that had been in contact with the skeletal remains. When comparing genetic profiles, we matched 28 of the 84 bones analyzed with living relatives (brothers, sisters, sons, daughters, nephews, or cousins). The statistical analyses showed a high confidence of correct identification for all 28 victims in the Konfin I mass grave (posterior probability ranged from 99.9% to more than 99.999999%).     

Bringing colour back after 70 years: Predicting eye and hair colour from skeletal remains of World War II victims using the HIrisPlex system

Retrieving information about externally visible characteristics from DNA can provide investigative leads to find unknown perpetrators, and can also help in disaster victim and other missing person identification cases. Aiming for the application to both types of forensic casework, we previously developed and forensically validated the HIrisPlex test system enabling parallel DNA prediction of eye and hair colour. Although a recent proof-of-principle study demonstrated the general suitability of the HIrisPlex system for successfully analysing DNA from bones and teeth of various storage times and conditions, practical case applications to human remains are scarce. In this study, we applied the HIrisPlex system to 49 DNA samples obtained from bones or teeth of World War II victims excavated at six sites, mostly mass graves, in Slovenia. PCR-based DNA quantification ranged from 4 pg/μl to 313 pg/μl and on an average was 41 pg/μl across all samples. All 49 samples generated complete HIrisPlex profiles with the exception of one MC1R DNA marker (N29insA) missing in 83.7% of the samples. In 44 of the 49 samples (89.8%) complete 15-loci autosomal STR (plus amelogenin) profiles were obtained. Of 5 pairs of skeletal remains for which STR profiling suggested an origin in the same individuals, respectively, 4 showed the same HIrisPlex profiles and predicted eye and hair colours, respectively, while discrepancies in one pair (sample 26 and 43) are likely to be explained by DNA quantity and quality issues observed in sample 43. Sample 43 had the lowest DNA concentration of only 4 pg/μl, producing least reliable STR results and could be misleading in concluding that samples 43 and 26 originate from the same individual. The HIrisPlex-predicted eye and hair colours from two skeletal samples, suggested to derive from two brothers via STR profiling together with a living sister, were confirmed by the living sister's report. Overall, we demonstrate that after more than 70 years, HIrisPlex-based eye and hair colour prediction from skeletal remains is feasible with high success rate. Our results further encourage the use of the HIrisPlex system in missing person/disaster victim identification to aid the identification process in cases where ante-mortem samples or putative relatives are not directly available, and DNA predicted eye and hair colour information provides leads for locating them, allowing STRbased individual identification.     

Digging up the recent Spanish memory: genetic identification of human remains from mass graves of the Spanish Civil War and posterior dictatorship

The Spanish Civil War (1936–1939) and posterior dictatorship (until 1970s) stands as one of the major conflicts in the recent history of Spain. It led to nearly two hundred thousand men and women executed or murdered extra-judicially or after dubious legal procedures. Nowadays, most of them remain unidentified or even buried in irretraceable mass graves across Spain. Here, we present the genetic identification of human remains found in 26 mass graves located in Northern Spain. A total of 252 post-mortem remains were analyzed and compared to 186 relatives, allowing the identification of 87 victims. Overall, a significant success of DNA profiling was reached, since informative profiles (≥12 STRs and/or mitochondrial DNA profile) were obtained in 85.71% of the remains. This high performance in DNA profiling from challenging samples demonstrated the efficacy of DNA extraction and amplification methods used herein, given that only around 14.29% of the samples did not provide an informative genetic profile for the analysis performed, probably due to the presence of degraded and/or limited DNA in these remains. However, this study shows a partial identification success rate, which is clearly a consequence of the lack of both appropriate family members for genetic comparisons and accurate information about the victims’ location. Hence, further perseverance in the exhumation of other intact graves as well as in the search of more alleged relatives is crucial in order to facilitate and increase the number of genetic identifications.     

The identification of war victims by reverse paternity is associated with significant risks of false inclusion

Since February 2001 the process of DNA identification of war victims in Croatia relies on the database of over 3,000 9-locus (D3S1358, vWA, FGA, TH01, TPOX, CSF1PO, D5S818, D13S317 and D7S820) STR genotypes of relatives of missing persons. Instead of a targeted approach to DNA typing, the genotype of each skeletal remains analysed is compared to all genotypes in the database to identify potential parents and children. Although this approach has significantly increased the pace of identification by DNA typing, non-targeted matching in a database containing several thousand genotypes considerably decreases the significance of inclusion, especially when identification is based on reverse paternity analysis. To support this statistical prediction we present 3 cases of 10 STR loci matches and 1 case of 11 STR loci matches between a child, child's mother and skeletal remains that did not originate from a father of that child.     

Use of pacemaker programmers for disaster victim identification

Disaster victim identification (DVI) presents a number of physical and legal challenges, involving the degeneration of human remains and legal obstacles to forensic examinations. One non-invasive method for positive identification may be the use of a pacemaker programmer to detect and obtain data from pacemakers recovered from unidentified remains. To test the usefulness of this method, this investigation examined the efficiency and utility of 5 different pacemaker programmers in the positive identification of victims of the March 2011 tsunami in Japan at 8 disaster sites in May 2011. On scanning 148 sets of remains, data were successfully obtained from 1 implant in 1 set of remains, allowing for the rapid positive identification of the individual. Scanning pacemakers with pacemaker programmers can be a non-invasive method of positive identification that meets Japanese legal and institutional requirements, but this method is ineffective without a preceding whole-body X-ray scan.     

DNS-Laborstrategie zur Identifizierung von Katastrophenopfern

More than 400 samples from 101 Tsunami victims from Sri Lanka were sent to our laboratory. Barcoded sampling kits were used to collect 2 swabs from internal body regions, a 4 cm piece of long bone and 2 teeth, which were all shipped on ice. A high throughput DNA extraction and STR analysis procedure for the swabs was developed in order to provide STR profiles within 24 h. For the skeletal remains and teeth a highly sensitive DNA extraction procedure was applied Both processes included electronic accessioning to maintain the numbering system of the DVI team in Sri Lanka and data exchange via Interpol and Plass Data software. DNA profiling of swabs was successful in 42% of the cases. For the remaining cases skeletal remains had to be typed and produced useful results in 65% for teeth and in 94% for bone. A sufficient profile was obtained from each victim, so the overall success rate was 100%. Until now DNA matching procedures in Innsbruck facilitated by using DNAVIEW software enabled DNA-based identification of 24 victims from 9 countries.     

Forensisch-odontologische Identifizierung der Opfer des Flugunfalls Zell am See

In 2007 the most serious aircraft accident in Austria since the Second World War occurred. The collision between a cargo helicopter and a light aircraft at a height of 800 m in the area of Zell am See near Salzburg resulted in 8 victims. The victims and the wreckage were spread over an area of approximately 20 hectares. The victim’s bodies were sent to the department of forensic medicine in Salzburg for autopsy and identification. The identification was carried out according to the Interpol guidelines for victim identification and involved the three primary identification markers fingerprints, DNA analysis and dental status. Of the eight victims six could be successfully identified by the dental status on the basis of antemortem (AM) dental charts. All victims could be identified by fingerprints and DNA analysis.     

Identifizierungstätigkeit bei Massenunfällen und Katastrophen

The identification of victims in cases of mass disasters is still a great challenge for all concerned. On the one hand an adaptive strategy is necessary for the identification workflow depending on the type of catastrophe and on the other hand high public pressure demands a fast identification process of missing relatives. The international cooperation after the Tsunami in 2004 showed under what conditions successful victim identification can be carried out. This led among other things to intensification of international cooperation at various levels, which is necessary in order to attain basic standards for identification for all participating countries. The identification commission (IDKO) of the Bundeskriminalamt (BKA) has actively participated in this international cooperation in recent years with support from forensic pathology, forensic odontostomatology and forensic molecular genetics. In this article the established standards and training options in the German victim identification process will be reported.     

Disaster victim identification

In the event of any mass fatality incident, despite the cause, disaster victim identification must be undertaken; the humanitarian and legal responsibility for this falls on the forensic community. Mass fatality incidents can be natural (e.g., tsunamis, earthquakes, hurricanes), accidental (e.g., building collapse, ship sinking) or can occur as a result of a terrorist attack. Terrorism alone has been responsible for thousands of deaths in recent years and can be encountered in many forms (e.g., suicide bombings, airplane hijackings). In mass fatality situations, the experitise of many specialities are called on to assist in the identification efforts and to allow for the speedy return of recovered human remains to the relatives of the deceased. Today, DNA plays a vital but never solitary role in disaster victim identification.     

Disaster victim identification of military aircrew, 1945-2002

BACKGROUND: Aviation accident fatalities are characterized by substantial tissue disruption and fragmentation, limiting the usefulness of traditional identification methods. This study examines the success of disaster victim identification (DVI) in military aviation accident fatalities in the Australian Defense Force (ADF). METHODS: Accident reports and autopsy records of aircrew fatalities during the period 1945-2002 were examined to identify difficulties experienced during the DVI process or injuries that would prevent identification of remains using non-DNA methods. RESULTS: The ADF had 301 aircraft fatalities sustained in 144 accidents during the period 1945-2002. The autopsy reports for 117 fatalities were reviewed (covering 73.7% of aircrew fatalities from 1960-2002). Of the 117 victims, 38 (32.4%) sustained injuries which were severe enough to prevent identification by traditional (non-DNA) comparative scientific DVI techniques of fingerprint and dental analysis. CONCLUSIONS: Many of the ADF fatalities who could not be positively identified in the past could be identified today through the use of DNA techniques. Successful DNA identification, however, depends on having a reference DNA profile. This paper recommends the establishment of a DNA repository to store reference blood samples to facilitate the identification of ADF aircrew remains without causing additional distress to family members.     

山东济宁辐射事故受照人员生物剂量估算及彗星电泳检测分析

目的探索特大剂量照射后外周血和骨髓染色体培养方法，拟合6Gy以上大剂量照射染色体双着丝点＋环剂量-效应曲线，对山东济宁“10．21”事故受照者进行准确生物剂量估算和DNA损伤检测。方法采集2例受照者外周血和骨髓细胞，制备染色体标本，计数双（多）着丝点＋环数目；用正常离体人血拟合6～22Gy双＋环剂量效应曲线及数学方程；对2例事故受照者进行生物剂量估算。用碱性单细胞凝胶电泳方法检测受照者外周血DNA损伤。结果B的外周血染色体双＋环平均数为4．47个／细胞；A的外周血培养无分裂细胞，骨髓染色体双＋环平均数为9．15个／细胞。用6—22Gy剂量效应方程估算全身平均受照剂量，B为9．4Gy，A为19．5Gy。单细胞凝胶电泳可见2例受照者的多数彗星细胞呈小头大尾形状。结论用新建立的6～22Gy染色体畸变剂量效应曲线估算2例受照者的生物剂量，已分别达到极重度骨髓型放射病和肠型放射病水平。     

Y-STR DNA analysis of 154 female child sexual assault cases in the Philippines

The laboratory evaluated 154 sexual assault cases from four Child Protection Units in the Philippines involving female child victims aged from 2?years to 18?years old. All child victims sought medical attention within 72?h after sexual contact. In 130 cases, the child victim knew the alleged offender and identified them during the interview with the social worker. Penile ejaculation was reported by 68 child victims with varying reports of washing after contact. Overall, 84 child victims admitted having wiped their genitalia prior to the collection of biological samples for DNA testing. Laboratory personnel examined vaginal smears in only 109 cases using a light microscope and reported 23 samples to be positive for sperm cells. Using the PowerPlex? short tandem repeat of the Y chromosome (Y-STR) DNA multiplex system, male DNA was detected in vaginal swab samples from 63 child victims. In 39 cases, positive amplification at 11 Y-STR DNA markers consistent with a single male DNA profile was observed. Twenty-eight of these full single Y-STR DNA profiles were found to be unique when searched in worldwide Y-STR DNA population databases (~40,000 haplotypes), eight haplotypes matching Filipinos and/or Asian haplotypes and one Y-STR DNA profile only matching European, Caucasian, and Latin American haplotypes. Y-STR DNA profiles generated will be compared with reference DNA profiles of alleged offenders once reference samples are submitted to the laboratory.     

Streamlining the decision-making process for international DNA kinship matching using Worldwide allele frequencies and tailored cutoff log10LR thresholds

The identification of human remains belonging to missing persons is one of the main challenges for forensic genetics. Although other means of identification can be applied to missing person investigations, DNA is often extremely valuable to further support or refute potential associations. When reference DNA samples cannot be collected from personal items belonging to a missing person, a direct DNA identification cannot be carried out. However, identifications can be made indirectly using DNA from the missing person’s relatives. The ranking of likelihood ratio (LR) values, which measure the fit of a missing person for any given pedigree, is often the first step in selecting candidates in a DNA database. Although implementing DNA kinship matching in a national environment is feasible, many challenges need to be resolved before applying this method to an international configuration. In this study, we present an innovative and intuitive method to perform international DNA kinship matching and facilitate the comparison of DNA profiles when the ancestry is unknown or unsure and/or when different marker sets are used. This straightforward method, which is based on calculations performed with the DNA matching software BONAPARTE, Worldwide allele frequencies and tailored cutoff log₁₀LR thresholds, allows for the classification of potential candidates according to the strength of the DNA evidence and the predicted proportion of adventitious matches. This is a powerful method for streamlining the decision-making process in missing person investigations and DVI processes, especially when there are low numbers of overlapping typed STRs. Intuitive interpretation tables and a decision tree will help strengthen international data comparison for the identification of reported missing individuals discovered outside their national borders.     

Forensic aspects of mass disasters: strategic considerations for DNA-based human identification

Many mass disasters result in loss of lives. Law enforcement and/or public safety and health officials often have the responsibility for identifying the human remains found at the scene, so they can be returned to their families. The recovered human remains range from being relatively intact to highly degraded. DNA-based identity testing is a powerful tool for victim identification in that the data are not restricted to any particular one to one body landmark comparison and DNA profile comparisons can be used to associate separated remains or body parts. Even though DNA typing is straightforward, a disaster is a chaotic environment that can complicate effective identification of the remains. With some planning, or at least identification of the salient features to consider, stress can be reduced for those involved in the identification process. General guidelines are provided for developing an action plan for identification of human remains from a mass disaster by DNA analysis. These include: (1) sample collection, preservation, shipping and storage; (2) tracking and chain of custody issues; (3) laboratory facilities; (4) quality assurance and quality control practices; (5) parsing out work; (6) extraction and typing; (7) interpretation of results; (8) automation; (9) software for tracking and managing data; (10) the use of an advisory panel; (11) education and communication; and (12) privacy issues. In addition, key technologies that may facilitate the identification process are discussed, such as resin based DNA extraction, real-time PCR for quantitation of DNA, use of mini-STRs, SNP detection procedures, and software. Many of the features necessary for DNA typing of human remains from a mass disaster are the same as those for missing persons' cases. Therefore, developing a missing persons DNA identification program would also provide the basis for a mass disaster human remains DNA identification program.     

Theoretical value of the recommended expanded European Standard Set of STR loci for the identification of human remains

We have undertaken a series of simulations to assess the effectiveness of commercially available sets of STR loci, including the loci recommended for inclusion in the expanded European Standard Set, for the purpose of human identification. A total of 9200 genotype simulations were performed using DNA · VIEW. The software was used to calculate likelihood ratios (LRs) for 23 groups of relatives, and to determine the probability of identification given scenarios that ranged between 10 and 250,000 victims. The additional loci included in the recommended expanded European Standard Set, when used in conjunction with the Identifiler(?) kit, significantly improved the typical LRs for tested scenarios and the likely success of providing correct identifications.     

Drug and alcohol use by homicide victims in Trinidad and Tobago, 2001–2007

This paper examines toxicology results from homicide victims in Trinidad and Tobago to explore patterns in pre-mortem drug and alcohol use. Toxicology test results were obtained for 1,780 homicide victims. Toxicology data from the coroner's office were linked with police data on homicide incidents to examine patterns in drug use and homicide. Trinidad and Tobago homicide victims tested positive for cannabis at a significantly higher rate (32%) than the average rate among other drug toxicology studies. Victims tested positive for alcohol (29%), cocaine (7%), and opioids (1.5%) at rates that were either comparable with or lower than those of homicide victims examined in other studies. The proportion of victims testing positive for cannabis grew significantly from 2001 to 2007; the proportions for alcohol and other drugs were fairly stable over time. Toxicology results also varied by homicide motive, weapon type, and the demographic characteristics of the victim. Toxicology data are a useful source for understanding patterns in drug use and homicide. Though such data have limitations, when combined with other types of data, they can often provide unique insights about a community's drug and violence problems.     

Evaluating the prevalence of DNA mixtures found in fingernail samples from victims and suspects in homicide cases

An important aspect of homicide investigations is the identification of the persons that had the last contact with the victim prior to death. Violent crimes are frequently characterized by a struggle between the victim and the perpetrator where biological material can be expected to be exchanged between them.Forensic DNA typing enables the generation of genetic profiles by extraction and amplification of cellular material found under fingernails. The evidential value of these samples may be critical if the secondary contributor found in a DNA mixture, can be matched with a potential suspect, or through a DNA database search.The amount of biological material transferred under the fingernails during “casual” activities is not sufficient to genotype reportable mixtures. This may not be the case with homicide victims that may have struggled and died under violent circumstances.The aim of this study was to evaluate the prevalence of DNA mixtures found under the fingernails of both victims and suspected perpetrators of violent deaths.We present a retrospective study of 137 DNA profiles genotyped from fingernail samples of homicide victims and suspects, collected at the Israeli National Center of Forensic Medicine. The majority of the samples produced single source profiles (n = 107, 78%) that matched those of the donor's. DNA mixtures (n = 30, 22%) were found in increased frequency among victims (n = 25/100, 25%) compared to suspects (n = 5/37, 13.5%). Mixtures were sub-divided into high level (n = 15, 50%), low level (n = 9, 30%) and residual (n = 6, 20%), according to the number of the foreign contributors’ alleles. Thus, this distinctive group of homicide victims was found to express both elevated frequency of DNA mixtures together with highly informative value of the secondary foreign profiles, as compared to other studied populations. These findings support an important aspect for the criminal investigation in murder cases, where a struggle may have ensued and the identification of an additional profile found in a mixture from a fingernail sample may point to a possible perpetrator of the crime.     

Loss of gray–white matter discrimination as an early CT sign of brain ischemia/hypoxia in victims of asphyxial cardiac arrest

Brain CT obtained from cardiac arrest (CA) victims immediately after resuscitation may be useful in predicting their outcomes. Most data have been derived from CA victims of cardiac etiology, however, CT signs of brain ischemia/hypoxia have rarely been studied in victims of asphyxial CA. Loss of gray–white matter discrimination (GWMD) at the basal ganglia seems to be the most reliable early CT sign of brain ischemia/hypoxia; a retrospective study was conducted to clarify its incidence, prognostic significance, and temporal profile in resuscitated victims of CA by food asphyxiation. Brain CT scans of each victim were interpreted by two blinded observers. During a 5-year period, 39 resuscitated victims of CA by food asphyxiation underwent brain CT. Thirty-one (79%) showed loss of GWMD, none of whom survived to discharge. Among the other eight victims with seemingly intact brain CT, five (63%) survived to discharge. Loss of GWMD predicted fatality with sensitivity of 100% and specificity of 63%. The interobserver concordance was 82% with kappa coefficient of 0.56. Loss of GWMD developed almost invariably when the asphyxiation–return of spontaneous circulation (ROSC) interval exceeded 10 min. There were five victims with asphyxiation–ROSC interval ≤ 10 min, all of whom survived to discharge. In contrast, none of the 34 victims with the interval >10 min survived to discharge. Loss of GWMD may develop in a relatively time-dependent manner and may be a reliable radiographic indicator of poor outcome in resuscitated victims of asphyxial CA.     

Quality and quantity of DNA in cadavers' serum

We investigated the quality and quantity of DNA in sera obtained from 16 cadavers who were left at room temperature or placed in a refrigerator for cadavers within 3 days of their death. The causes of death were as follows: asphyxia, 4; hemorrhage, 4; fire, 3; cold, 1; subdural bleeding, 1; sepsis, 1; cerebral contusion, 1; and pulmonary thromboembolism, 1. When examined using agarose gel electrophoresis, the samples from the victims who died from sepsis and pulmonary thromboembolism exhibited smear patterns, while the other samples exhibited ladder patterns. The size of each band was an integer multiple of approximately 180 bp, considered to be characteristic of apoptosis. The DNA concentration in the cadavers' sera ranged from 3.0 to 258.9 ng/ml. DNA types (D1S80, HLADQA1, TH01, and PM) were determined for all the samples except for the sample from the victim who died from sepsis. The detected types were the same as those obtained from paired bloodstain samples, except for in the asphyxia victim. D1S80 typing of the serum sample from the asphyxia victim showed the loss of the longer allelic hetero-band. These results suggest that the quantity of DNA in serum from cadavers is sufficient for DNA typing, but that the degradation of DNA may proceed quickly, depending on the temperature of the area in which the cadaver is left as well as the pathological conditions that the victim had experienced.