Application Of Forensic Dna Identification In Mass Disaster Investigations


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  • DMORT is a federal agency employing part time federal employees, not volunteers, who work as forensic professionals (pathologists, dentists, anthropologists, funeral directors, etc), not medical students
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  • -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: sample collection, preservation, shipping and storage; 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 (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.
  • Always collect the donor’s full name and date of birth. During times of grief, relatives may not realize that they are using nicknames or that a father’s “Bob” may be a mother’s “Robby.” ■ Europeans and Americans write dates differently (the standard European notation is DD/MM/YY). Ensure that month and day fields are unambiguous on collection forms. ■ Family members frequently transpose their relationship to the victim. In most cases, this is a result of a poorly worded question such as, “What is your relationship to the victim?” It is better to ask questions from the perspective of the donor. For example, “The victim is my ___________.” or “I am the victim’s ___________.” Also, the dates of birth of the donor and the victim can be used to help correct these mistakes. ■ Collect as much information as possible about the relevant family structure; the sample form found in appendix C may be a helpful guide. The laboratory can compare purported pedigrees from members of the same family, then use dates of birth and genotypes to help discern the true relationships. ■ Collect as much information and as many samples as possible. There may not be another opportunity.
  • -In a “closed” incident, the laboratory director should determine whether a list of victims is available—for example, in an airline disaster, the passenger manifest. Although it is important to keep in mind that the manifest might be incomplete or incorrect, the majority of the victims would still be known. -It is possible for a closed incident to become open. If a plane crashes into a neighborhood, for example, the victims on the ground would change a typical “closed” event to “open,” because it would not be known who was on the ground. -It should also be kept in mind that open incidents are prime candidates for insurance fraud. There are people who may try to file fraudulent life insurance claims.
  • - To help with this purpose, specialized and trained disaster victim identification (DVI) multidisciplinary teams composed of medical examiners, forensic pathologists, anthropologists, forensic odontologists, fingerprint specialists, radiologists, and experts in search and recovery of physical evidence have been developed worldwide.
  • -prepared the following recommendations to help the OCME laboratory create policies and procedures specific to the WTC mass fatality incident -provided a roadmap when it was necessary to depart from the laboratory’s usual forensic casework protocols -recommendations also offered guidance for securing additional resources and provided assurance that sufficient peer review and expertise were available to support these new endeavors -WTC CODIS [Combined DNA Index System] be used: — At high stringency for direct matches. Likelihood ratio of 1 X 1010 is sufficient to report identity. A 13-locus match using the core CODIS loci is sufficient to report identity. — At low stringency to screen for potential first-degree relatives (parent/offspring and some sibs) in order to manually search case-specific data for cases with additional potential relatives.
  • -As the scope of the WTC effort evolved, and the complexities of data management and the number of partnerships increased, the KADAP recommended and implemented a mechanism to facilitate secure, rapid transfer of data and provided additional development of statistical approaches to kinship analyses. -DNA•VIEW be used to assess the putative relationship. A minimum Probability of Relationship of 99.9% is sufficient to report identity by kinship analysis. The minimum prior probability is 1/5000, which can be increased to reflect case-specific issues (e.g., members of service). -Successful DNA typing of all samples will not be possible due to conditions of the remains. The Panel recommends that testing of individual samples should be finite. Criteria for determining cessation of testing should be established. Development of a probative test should be investigated. -Mitochondrial DNA typing of victim samples should be used only as a last resort after additional test reanalysis and/or the use of additional forensically validated STR, Y-chromosome, or other nuclear markers have been used. - If forensically validated systems, including mitochondrial data, are insufficient to resolve identity, research grade systems should be explored on a case-by-case basis.
  • -recommended to collect DNA samples from the least affected material to avoid exogenous and cross contamination skeletal muscle organ tissues skin blood -new system developed for bar-coded based soft tissue collection and body tracking
  • -Sea conditions are bad for the condition of DNA integrity - 9.0-magnitude earthquake off the coast of Sumatra sent several tsunami waves radiating out into the Bay of Bengal, the Andaman Sea, and the Indian Ocean
  • Tsokos, et al. : - hurried disposal of corpses by cremation or mass burials was performed in the region of Phuket and Khao Lak at some places under the surveillance of local authorities, resulting in bodies being either wrongly or not identified -Approximately 2,000 bodies that had been transported here from beaches, hotel resorts and inland, where they had been recovered by Thai military and civil volunteers, were lying in rows on the ground in the open -An immense problem was the lack of sufficient means of cooling the dead in order to slow down putrefaction; an ineffective attempt to slow down putrefaction and to preserve the bodies in their present state was made by the Thai military by cooling the corpses with dry ice
  • Tsokos, et al. : -Sex determination was not a problem in any of the bodies examined due to the presence of the reproductive organs; body parts were not among the human remains examined by the German DVI team. -Estimation of age was mainly based on odontological features, occasionally supported by anthropologic data such as skull suture fusion, epiphyseal union of distal radius and ulna, etc. -The causes of death of those tsunami victims examined by the German DVI team were exclusively drowning and blunt trauma. Yamada et al: - Handling bodies of the deceased. According to the Sri Lankan Judicial System, the physicians in charge of the district hospitals perform autopsies and other forensic pathologic duties. -Specialized judicial medical officers (forensic pathologists) are available only in larger provincial hospitals. -As such, it is customary to bring all dead bodies to the hospital when deaths occur outside the hospital. -Hospital morgues are usually equipped to handle fewer than 5 to 10 bodies.
  • -for expertise, management and support in performing the DNA identifications
  • Donkervoort, et al. : - In the identification efforts following Hurricane Katrina, a Family and/or Donor Reference Collection (FDRC) form was used as a means to document the reported relationship between the reference DNA donor and the purported missing individual. -This FDRC form was developed based upon lessons learned from 9/11 and the Tsunami identification efforts. -This paper analyses the effectiveness of the FDRC form used in the Hurricane Katrina kinship DNA identification efforts and proposes an improved sample collection form for kinship and other donor reference samples. -Between September 2005 and June 2006, there were approximately 1500 FDRC forms completed of which 724 were reviewed and analyzed for this study. -
  • Project Management ensures that all functions work in concert to provide accurate identifications as rapidly as possible within budgetary constraints. Sample Accessioning and Tracking consists of accessioning remains and reference samples, ensuring chain-of-custody documentation, and managing the flow of samples and data within the laboratory and among outsourced laboratories. Sample Analysis means performing DNA tests on remains and reference samples. Identification Management has two parts: (1) making identifications by matching remains and reference samples and (2) reviewing the metadata—information from all sources linked to a particular sample—associated with the reference samples to ensure they were correctly associated with the putative victim. Quality Control refers to the processes and procedures that a laboratory uses to detect and avoid mistakes. Quality control also ensures that there are no discrepancies between DNA analysis and other modalities (i.e., that the metadata can be reconciled when a conflict occurs). Information Technology (IT) and Informatics includes the software and hardware that supports the identification effort. Human Resources focuses on meeting the needs of the staff, volunteers, and consultants who are working on the response effort. Media Relations involves interacting with the press and establishing how and when information is released to the media. Family Coordination encompasses educating families, collecting the reference samples and family data necessary to identify victims, and providing information to the families. Procurement involves ensuring that the correct equipment, supplies, and services are available to the response in a timely manner.
  • - Experiences gained from previous mass fatality incidents reinforce the need to make all necessary steps to guarantee sample preservation for DNA analysis and to use suitable protocols for documenting the chain of custody of DNA sampling and body tracking.
  • -low rate of body recovery -
  • - In most disasters (Table 1), the standard of care is usually to identify each victim, but under certain conditions, like those after the WTC tragedy, with very high level of body fragmentation and sample disintegration from an open population, the challenge is to identify each remain by DNA testing (10,11). -Sample preference for DNA analysis is also determined by each mass disaster scenario. In general, it is recommended to collect samples from the least affected material in a way to avoid both exogenous contamination and body crosscontamination. -The preferable human remain sources include: soft tissues (skeletal muscle, organ tissues and skin) and blood.
  • -Software tools for storing, tracking, comparing, annotating and curating data had never been considered a priority in North America, because there was no previous case of the scale and scope of the WTC. -Most previous mass disasters had been relatively small scale and simple spreadsheet approaches were sufficient to organize and track data.
  • -Even when ante-mortem data is unavailable, M-FISys' kinship analysis lets family members know if remains collected from a disaster belong to their missing relatives. -In addition to forming a valuable addition to the emergency response repertoire on a national or local level, M-FISys is also able to handle data from large-scale civil incidents such as air crashes, where human remains are frequently fragmented. -Direct matching of DNA profiles to personal effects (references), with immediate access to raw electropherogram and image data for each sample. This ensures that trained Forensic Scientists always have access to the data they need to make accurate identifications -QA tools allowing you to spot inconsistencies and errors in the data that may be the result of commingled remains, sample switches or contamination and greatly reduce the chance of a false-identification.
  • Alonso, et al. : - high number of pair wise comparisons – from thousand to millions, or even more in the Tsunami catastrophe, which have to be carried out to correlate victims to direct and/or family references. -the significance of a genetic compatibility (allele sharing) between a sample of the victim’s index and a sample of the relative’s index can be challenged in mass disaster cases by the incidence of fortuitous hits (false positives) even in the case of parent-child relationships. Leclair et al: -Swissair Flight 111 disaster in 1998: 229 passengers and crew perished, the crash dispersed thousands of severely fragmented human remains -Successful PCR amplification produced genotypes for 97% of the recovered remains -some of the most useful personal effects of the victims had been lost at sea in luggage, this route, although useful, was not perceived as being sufficient. -Kinship analysis was considered, but both biological parents and/or offspring and spouse were available for only 26% of all victims -On the basis of Mendelian inheritance rules, suspected parent/offspring relationships could be inferred by computing the number of loci at which at least one allele is shared. -The scheme was automated with VBA in Excel. It permitted for all possible (i.e. 180,000) pair-wise genotype comparisons to be examined. -As a closed population of victims was reached for this tragedy, the approach also permitted for the systematic exclusion of fortuitous kinship associations.
  • -Multiplex PCR amp preferred since it is simple, sensitive, and has high discrimination power
  • Include OKC bombing How did 9/11 change what was done with the ID of the tsunami victims? What happened before 9/11? Did 9/11 have any effects on how we treated/apprached Katrina?
  • Application Of Forensic Dna Identification In Mass Disaster Investigations

    1. 1. The Application of Forensic DNA Identification in Mass Disaster Investigations Cheryl M. Lowe March 11, 2009
    2. 2. Introduction <ul><li>Identification of bodies in mass disaster cases most commonly by fingerprints and dental records </li></ul><ul><li>However, DNA analysis can be done as well…especially when such evidence is unavailable </li></ul><ul><li>DNA techniques can be used in almost any mass disaster investigation: crashes, natural (storms), accidents (fires), wars, bombings </li></ul>
    3. 3. Introduction <ul><li>Unfortunately most mass disasters are messy and chaotic, making identification efforts more difficult </li></ul><ul><li>Learning process for society each time a large-scale mass fatality occurs </li></ul><ul><ul><li>Organization and management </li></ul></ul><ul><ul><li>Speed and effectiveness of emergency response </li></ul></ul><ul><ul><li>Ethical issues </li></ul></ul><ul><ul><li>Quality control </li></ul></ul>
    4. 4. Mass Disasters Affect Everyone <ul><li>Important for family members to receive closure about a loved one </li></ul><ul><li>Victims’ families should be quickly informed about the processes behind the identification efforts </li></ul><ul><li>Budowle, et al. – there should be a positive supportive relationship with family members so that the collection of antemortem reference samples goes more smoothly </li></ul><ul><li>Must inform families of any privacy issues associated with the use of DNA samples </li></ul>
    5. 5. Relationship Between Laboratory and Victims’ Families
    6. 6. Types of Mass Disasters <ul><li>Crashes: airplane, train, building collapses </li></ul><ul><ul><li>High degree of body fragmentation </li></ul></ul><ul><li>Natural: hurricanes, tsunamis, tornados, earthquakes, floods, other storms </li></ul><ul><li>Wars and bombings </li></ul>
    7. 7. Types of Mass Disasters <ul><li>“Closed” mass disasters = number and identity of victims more readily known </li></ul><ul><ul><li>Airplane crashes – typically have a list of passengers already available </li></ul></ul><ul><li>“Open” mass disasters = the number and identity of victims involved is mainly unknown </li></ul><ul><ul><li>World Trade Center tragedy – final list of victims undetermined until months after </li></ul></ul>
    8. 8. Key Players in Mass Disaster Identification Efforts <ul><li>Medical examiners </li></ul><ul><li>Forensic odontologists </li></ul><ul><li>Fingerprint specialists </li></ul><ul><li>Radiologists </li></ul><ul><li>Experts in search and recovery of physical evidence </li></ul><ul><li>Forensic anthropologists </li></ul><ul><li>Forensic pathologists </li></ul><ul><li>Forensic DNA analysts </li></ul> Communication
    9. 9. Lessons Learned: 9/11 Terrorist Attacks <ul><li>Each match from the World Trade Center disaster made by the private labs verified by the medical examiner’s lab. Police then inform families in person. </li></ul><ul><li>KADAP (Kinship and Data Analysis Panel) formed to assist the NYC Chief Medical Examiner’s office during the World Trade Center Tragedy </li></ul><ul><li>Gonzales, et al. – KADAP recognized the complexity of the 9/11 mass disaster incident, and proposed recommendations for procedures and protocols specific to the WTC incident </li></ul><ul><li>KADAP recommended that CODIS be used: </li></ul><ul><ul><li>at high stringency for direct matches </li></ul></ul><ul><ul><li>at low stringency to screen for possible first-degree relatives (parents, children, sibs) </li></ul></ul>
    10. 10. Lessons Learned: 9/11 Terrorist Attacks <ul><li>Gonzales, et al. -- Kinship used to confirm a personal effect match should be accepted at a Probability of Relationship of 99.9% using a Prior Probability of 0.5. </li></ul><ul><li>Understood that all samples cannot be successfully typed due to the conditions of the remains; KADAP recommended criteria for ending testing of individual samples </li></ul><ul><li>mtDNA analysis used as a last resort; should be performed on all maternal lineage relative’s appropriate samples (e.g., buccal swabs, blood) using a suitable validated system </li></ul>
    11. 11. Lessons Learned: Southeast Asian Tsunami <ul><li>Alonso, et al. – more than 200,000 victims total; victims from a span of 30 countries </li></ul><ul><li>Disaster victim identification (DVI) teams organized </li></ul><ul><li>High levels of body fragmentation and high rate of DNA degradation </li></ul><ul><li>Many of the bodies were inaccessible </li></ul><ul><li>Many entire families perished, making kinship analysis efforts difficult if not impossible </li></ul>
    12. 12. The Tsunami’s Effect
    13. 13. Lessons Learned: Southeast Asian Tsunami Tsokos M, Lessig R, Grundmann C, Benthaus S, Peschel O. Experiences in tsunami victim identification. Int J Legal Med 120 (3) (2006) 185-187.
    14. 14. Tsunami Tsokos M, Lessig R, Grundmann C, Benthaus S, Peschel O. Experiences in tsunami victim identification. Int J Legal Med 120 (3) (2006) 185-187.
    15. 15. Lessons Learned: Hurricane Katrina <ul><li>August 29, 2005—Hurricane Katrina struck the Gulf Coast of the United States </li></ul><ul><li>Catastrophic flooding throughout the majority of the New Orleans, LA region </li></ul><ul><li>Over 12,000 individuals reported missing and an estimated 1336 fatalities (Donkervoort, et al.) </li></ul><ul><li>In Louisiana, medical and dental records were destroyed and the condition of human remains following prolonged water and heat exposure impaired visual identification </li></ul>
    16. 16. Lessons Learned: Hurricane Katrina <ul><li>The Louisiana Department of Health and Hospitals was responsible for the identification of victims </li></ul><ul><li>Utilized the Louisiana State Police Crime Laboratory DNA Unit </li></ul><ul><li>Many personal items from the victims such as toothbrushes were lost or destroyed, making the use of direct DNA identification impossible </li></ul>
    17. 17. Hurricane Katrina
    18. 18. Hurricane Katrina
    19. 19. Management of Mass Disaster Investigations <ul><li>A project manager should be assigned to create a schedule, assign resources, and monitor progress </li></ul><ul><li>Gonzalez, et al. – Certain aspects of DNA identification may not be understood by other groups working with the lab, so it is imperative that extensive communication occur </li></ul><ul><li>Anticipate and prepare for any amount of workload and possible problems or limits to the investigation </li></ul>
    20. 20. Management of Mass Disaster Investigations
    21. 21. Project Mgmt. Graph
    22. 22. DMORT <ul><li>Disaster Mortuary Operational Response Team </li></ul><ul><li>Group of professional volunteers, usually medical students, that are responsible for helping retrieve samples from the morgue </li></ul><ul><li>Developed the Victim Identification Program (VIP), software that helps to collect victim information </li></ul>
    23. 23. DMORT
    24. 24. Current Challenges <ul><li>Alonso, et al. – high number of victims, body destruction, DNA degradation, often limited reference samples available </li></ul><ul><li>Decreased number of PCR markers available </li></ul><ul><li>Remains from fire accidents can by extremely charred and indistinguishable </li></ul><ul><li>Lack of technical resources </li></ul><ul><li>Low success rate with STR loci </li></ul>
    25. 25. Current Challenges Alonso A, Martin P, Albarran C, Garcia P, Fernandez de Simon L, Iturralde MJ, et al. Challenges of DNA profiling in mass disaster investigations. Croat. Med. J 46(4) (2005) 540-548.
    26. 26. Current Challenges <ul><li>Alonso, et al. -- recommended a procedure for reducing errors during data collection is the use of specific and standardized sample collection forms employing a unique numbering systems to identify each remain in conjunction with the use of </li></ul><ul><li>Laboratory Information Management Systems (LIMS) = ensure sample information logging on a centralized database </li></ul>
    27. 27. Data Sorting <ul><li>Budowle, et al. – Software used in the investigation must: </li></ul><ul><ul><li>organize, store, and retrieve diverse and different data </li></ul></ul><ul><ul><li>integrate different software systems </li></ul></ul><ul><ul><li>allow technical and administrative review of data </li></ul></ul><ul><ul><li>allow for annotation and recording of problems and resolutions </li></ul></ul><ul><ul><li>track samples among partner laboratories </li></ul></ul><ul><ul><li>prioritize sample selection and review </li></ul></ul>
    28. 28. Data Sorting <ul><li>In addition, the software should also be able to: </li></ul><ul><ul><li>generate family pedigrees and calculate likelihood ratios for hypothesized kinships </li></ul></ul><ul><ul><li>combine remains with the same profile to facilitate searching </li></ul></ul><ul><ul><li>enable profile comparisons and statistical calculations </li></ul></ul><ul><ul><li>be user friendly </li></ul></ul>
    29. 29. M-FISys <ul><li>Mass-Fatality Identification System </li></ul><ul><li>Developed in response to WTC tragedy by Gene Codes Corporation </li></ul><ul><li>Able to make sense of huge quantities of data and to assist in the task of victim identification </li></ul>
    30. 30. Kinship Analysis <ul><li>Mass disasters usually have a high number of pairwise comparisons, ranging from 1,000s to millions </li></ul><ul><li>Software developed to: </li></ul><ul><ul><li>Collapse profiles from remains </li></ul></ul><ul><ul><li>Build overlapping partial profiles </li></ul></ul><ul><ul><li>Calculate likelihood ratios (LR) for each pairwise comparison at various relationships </li></ul></ul>
    31. 31. Kinship Analysis Alonso A, Martin P, Albarran C, Garcia P, Fernandez de Simon L, Iturralde MJ, et al. Challenges of DNA profiling in mass disaster investigations. Croat. Med. J 46(4) (2005) 540-548.
    32. 32. Future Mass Disaster Investigations <ul><li>As mass fatalities are unexpected and unpredictable, it is important to properly plan and learn from past experiences </li></ul><ul><li>Requires guidelines, training, preparation, and proper execution in order to be conducted efficiently </li></ul><ul><li>Alonso, et al. -- reduce errors during data collection by using specific, standardized sample collection forms with unique numbering systems to identify each remain </li></ul><ul><li>Multiplex PCR amplification preferred </li></ul><ul><li>Automation, robotic systems are helping to decrease costs and turnaround time </li></ul>
    33. 33. References <ul><li>Budowle B, Bieber FR, Eisenberg AJ. Forensic aspects of mass disasters: strategic considerations for DNA-based human identification, Leg. Med. (Tokyo) 7 (4) (2005) 230-243. </li></ul><ul><li>Zehner R. “Foreign” DNA in tissue adherent to compact bone from tsunami victims. Forensic Science International: Genetics 1 (2007) 218-222. </li></ul><ul><li>Morgan OW, Sribanditmongkol P, Perera C, Sulasmi Y, Van Alphen D, et al. Mass fatality management following the South Asian tsunami disaster: case studies in Thailand, Indonesia, and Sri Lanka. PLoS Medicine Vol. 3, No. 6, e195 doi:10.1371/journal.pmed.0030195 </li></ul><ul><li>Alonso A, Martin P, Albarran C, Garcia P, Fernandez de Simon L, Iturralde MJ, et al. Challenges of DNA profiling in mass disaster investigations. Croat. Med. J 46(4) (2005) 540-548. </li></ul><ul><li>Brenner, CH. Some mathematical problems in the DNA identification of victims in the 2004 tsunami and similar mass fatalities. Forensic Science International 157 (2006) 172-180. </li></ul><ul><li>Prinz M, Carracedo A, Mayr WR, Morling N, Parsons TJ, Sajantila A, et al. DNA Commission of the International Society for Forensic Genetics (ISFG): recommendations regarding the role of forensic genetics for disaster victim identification (DVI). FSI: Genetics (2007) 3-12. </li></ul><ul><li>Clayton TM, Whitaker JP, Maguire CN. Identification of bodies from the scene of a mass disaster using DNA amplification of short tandem repeat (STR) loci. Forensic Science International 76 (1995) 7-15. </li></ul><ul><li>Brenner CH, Weir BS. Issues and strategies in the DNA identification of World Trade Center victims. Theoretical Population Biology 63 (2003) 173-178. </li></ul><ul><li>Sorensen E, Hansen SH, Eriksen B, Morling N. Application of thiopropyl sepharose 6B for removal of PCR inhibitors from DNA extracts of a thigh bone recovered from the sea. Int J Legal Med (2003) 117: 245-247. </li></ul><ul><li>Graw M, Weisser HJ, Lutz S. DNA typing of human remains found in damp environments. Forensic Science International 113 (2000) 91-95. </li></ul>
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    37. 37. Questions?