Forensic Sci Med Pathol (2012) 8:140147

DOI 10.1007/s12024-011-9276-z

REVIEW

DNA analysis in disaster victim identification

Kerstin Montelius Bertil Lindblom

Accepted: 12 August 2011 / Published online: 19 October 2011

Springer Science+Business Media, LLC 2011

Abstract DNA profiling and matching is one of the primary methods to identify missing persons in a disaster, as
defined by the Interpol Disaster Victim Identification
Guide. The process to identify a victim by DNA includes:
the collection of the best possible ante-mortem (AM)
samples, the choice of post-mortem (PM) samples, DNAanalysis, matching and statistical weighting of the genetic
relationship or match. Each disaster has its own scenario,
and each scenario defines its own methods for identification of the deceased.
Keywords Disaster victim identification
DNA
identification
Mass disaster
Missing person
DNA
DVI

Introduction
DNA profiling and matching is one of the primary methods
used to identify missing persons in a disaster. Together
with odontology and dactyloscopy, the use of DNA is
recommended by the Interpol guidelines [1]. The process
of identifying a victim by DNA includes the collection of
the best possible ante-mortem (AM) samples, the choice of
post-mortem (PM) samples, and DNA analysis and statistical weighting of the genetic relationship or match.
Catastrophes can result from different scenarios such as
natural disasters, terror attacks, armed conflicts or accidents. Each scenario defines its identification methods and
samples selected. One of the first times DNA analysis was

K. Montelius (&)
B. Lindblom

National Board of Forensic Medicine,
Department of Forensic Genetics and Forensic Toxicology,
Artillerigatan 12, 587 58 Linkoping, Sweden
e-mail: kerstin.montelius@rmv.se

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used to identify mass disaster victims was after the fire on

the Scandinavian Star ferry in 1990. Here, restriction
fragment length polymorphism (RFLP) and analysis of
variable number of tandem repeats (VNTR) polymorphisms were used to identify the victims. A representative
review of mass disaster cases investigated by DNA profiling is given by Antonio Alonso et al. [2]. They described,
among other cases, 141 victims from an aircraft disaster in
Spitsbergen in 1996 [3] and the 9/11 World Trade Center
disaster, in which it was crucial to use DNA as method for
identification. The impact of the aircrafts hitting the World
Trade Centre (WTC) on 11 September 2001 generated
fragments from the victims which were only able to be
identified by DNA profiling. A large international operation followed the South East Asian tsunami where the
cooperation between different professions and disciplines
was a big challenge. The Disaster Victim Identification
(DVI) response to the South East Asian Tsunami was
evaluated by the Interpol Tsunami Evaluation Working
Group resulting in a series of recommendations [4]. Once
again DNA identification was an integral part of the
response.
This review intends to focus on experiences from
DVI-operations from a geneticists point of view.

Ante mortem samples

The Interpol DVI guidelines recommend three sources of
DNA AM data in the following order: (1) First degree
relatives, if possible more than one; (2) Blood or biopsy
samples from the potential victim; or (3) Personal objects
that have been used by the deceased. Samples from relatives can be conveniently collected on FTA-cards, either as
buccal swabs or blood samples.

Forensic Sci Med Pathol (2012) 8:140147

Two first degree relatives usually provide very good

data for identification. For example, two biological parents
or a child and one biological parent give informative
identifications [5]. Problems can arise when the victims are
relatives and comingled at the site of the disaster, as
described in a secluded village where many related people
died in a Taiwanese landslide [6]. Also, it is not possible to
distinguish siblings of the same gender on the basis of
DNA from their parents. The DNA analysis will tie the
siblings to the correct parents and family, but will not be
able to tell them apart.
The International Society for Forensic Genetics (ISFG)
guidelines suggest that a scientist with a genetic background should be available to provide advice on the collection of appropriate AM samples [7]. Relationship
information and chain of custody details must accompany
the samples. One approach to ensure all information is
collected is to use the Interpol DVI forms together with a
pedigree. Forms for both AM and PM sample collection
were developed for use in the identification process after
Hurricane Katrina [8]. Over all, forms were considered an
efficient way of collecting necessary, accurate data. However, the pedigree included was correctly completed in
only 72% of the cases. This means that staff had to go back
to gather important information in 28% of the cases. This
was time consuming and costly. It was suggested that the
relationship details as provided by the person be documented by the sample collector, and for the geneticist to
convert this information into a pedigree. Geneticists found
the pedigree very useful.
A pamphlet with appropriate genetic and contact information supports both the sample collectors in their work
and the relatives donating a sample.
The second option Interpol suggests as a source of AM
DNA data is bio bank samples. Bio bank samples could
be either medical specimens or bloodstains on filter paper
cards, intended for medical screening purposes. Such
specimens are kept in several countries, for example in
Australia and Sweden, where all newborns have been
screened for metabolic diseases for more than 30 years
[9]. The advantage of bio bank samples is that they are
labelled with the identity of the sample donor and kept in
secured banks in controlled conditions. And, analysis of
bio bank enables direct comparison to the DNA-profile of
the victims. In Australia the State Coroner can request the
Guthrie cards be made available for identification. In
Sweden, the use of bio bank samples has been restricted
to screening and research purposes only. During the
identification process for the South East Asian tsunami
victims, a temporary law was created, making the PKUcards available for this specific purpose until 30 June
2006. Fifteen Swedish victims were identified using PKUcards and one possible match between a victim and an

141

alleged mother was excluded. There is now a move to

make bio bank samples permanently available for identification in Sweden [10].
The third option for AM DNA samples recommended
by Interpol is personal objects used by the deceased. As
with bio bank samples, profiles generated from personal
objects can be directly compared to the DNA-profiles of
victims. The disadvantage is that there is a risk of mistakes
regarding who has owned or handled the object, leading to
false exclusions or misidentifications. Other items might
have been shared, giving mixed DNA-profiles as a result.
Baby teeth could also be considered as personal objects.
Analysis of a 60 year old baby tooth, showed a partial
profile of eleven out of fifteen markers. Baby teeth kept in a
small box in room temperature for 1 year have been shown
to provide very good DNA-profiles (Montelius et al. 2010
unpublished data). A classification of direct samples is
shown by Prinz et al. [7].

Post mortem samples

The circumstances of the disaster determine the method of
handling the bodies and collection of samples. Information
critical to body identification may be lost if the initial
process does not follow accepted DVI standards such as the
Interpol guidelines and ISFG recommendations [11, 12].
Proper labeling, documentation and chain-of-custody
are crucial, and several kits to assist with sample collection have been developed. Authorities in Norway and
Sweden have kits with various tubes, scalpels, labels and
other items, that might be needed during collection. Kits
with a radio frequency identification device have also
been tried [13]. Great Britain has developed a set of
documents including labels with PM numbers based on
the country code (personal communication). Each body,
as well as body parts, must be labeled with separate PM
numbers.
On Saturday the 24 June 2000, a small air plane crashed
in the southern part of Sweden. Four persons were reported
missing. A couple of weeks later, 53 PM samples arrived in
the department together with AM samples collected from
close relatives of the alleged victims. All 53 PM samples
were analyzed together with the AM samples using six
common markers available at the department at the time.
Summarizing the work, it was clear that it had been sufficient to analyze a few of the PM-samples to identify the
four victims. This DVI-work highlighted the question
regarding how many samples should be analyzed when
there are several victims and several body parts. It should
be decided at an early stage if all body parts are to be
analyzed and repatriated or if the work is finished when all
reported victims are identified.

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The rate of DNA degradation is influenced by many

factors including temperature, humidity, chemicals, (e.g.
formamide) and UV-radiation. There are several ways to
preserve the samples. Samples should be kept cool but if
that is not possible, DNA-preservatives such as Genofix
[13], SampleMatrix [14], regular salt [15], or alcohol,
can be used. If regular 95% alcohol isnt available,
commercial alcohol, such as white rum, has been proven
to be an adequate substitute [16]. Depending on circumstances and the condition of the body, the choice of
which sample to collect will also vary. DNA is preserved
in bone cells as well as in teeth (molars preferably).
Bone samples from different parts of the skeleton perform differently. The success rate of DNA profiles from
a selection of skeletal samples in the World Trade
Center disaster 2001 is reported by Mundorff et al. [17].
They concluded that samples from femur and metatarsal
bones work well, while skull bones are less suitable. For
less degraded remains, muscle tissue can be used [7].
Examples of PM-samples suitable for analysis are presented in Table 1.

Contamination risks
Contamination of the sample with DNA from other sources
is a risk. This could be from someone handling the sample,
contaminated instruments, or from comingled remains.
Guidelines for sample collection have been published [18]
with a series of suggestions on how to collect samples from
bone and teeth. The three main priorities are (1) protective
clothing and clean sampling areas, (2) clean instruments,
and (3) storage of samples in appropriate containers in
appropriate conditions. The authors also point out that
instruments might become blunt due to aggressive DNAdecontamination fluids, which is why disposable instruments are a better alternative. Caution has to be exercised
so as not to damage the sample when using decontaminants, such as bleach.
Precautions to avoid contamination during handling and
shipping of the samples from the site/morgue to the DNA
laboratories are also required. All samples need to be
individually wrapped in packing material, avoiding

Table 1 Recommendation for

collection of post-mortem
samples, adopted from Interpol
guide lines and ISFG guidelines

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leakage. Each sample needs to be labeled and have clear

chain of custody details.

DNA-analysis
Isolation of DNA
The method chosen for DNA-extraction is dependent on
the kind of sample and the protocols employed by the
laboratory. In a DVI situation there is little time to
implement new, un-validated methods. Examples of common PM samples are bone material, blood samples and
muscle tissue, and all require different extraction techniques. Extracted DNA should be quantified and preferably
checked for degradation.
Bio bank samples collected for medical purposes, are
often archival, formaline-fixed, paraffin embedded tissue. If
this kind of sample is intended to be used as an AM sample, it
is necessary to use an extraction method that is gentle enough
to preserve the DNA but at the same time efficiently removes
the paraffin. It has been shown that higher pH and higher
temperature during the extraction procedure produce better
results in terms of quality of DNA than more common
extraction protocols [19]. However, the alkali and high
temperature fragments the DNA, and if the sample is incubated for too long, there will be no DNA left.
AM samples collected on FTA cards can be analysed
using Polymerase Chain Reaction (PCR) directly without
extraction, decreasing the time required to obtain the DNA
profile.
DNA-profiling
Short Tandem Repeats (STR)
Fluorescent PCR multiplexes targeting STR loci are the
standard method used in human identification casework.
It is a prerequisite that AM samples and PM samples are
analyzed for the same loci. During the past year similar
kits for this purpose, from several suppliers, have been
launched. Even though the kits target the same loci,
differences in primer sequences means that different kits

Condition of body

Recommended sample

Complete non-decomposed body

Blood or buccal smears on FTA-cards

Mutilated non-decomposed body

Blood (if available) and deep red muscle tissue

Complete, decomposed corpse or mutilitated

remains
Severely burnt corpses

Compact bone or healthy teeth

Compact bone, healthy teeth or smears from internal
organs

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may amplify differing fragments. For this reason, it is

important to note what set of markers have been used for
DNA-profiling, as well as the kit that has been used.
Compromised samples may also result in allelic dropout.
Therefore, it is also important not to discard single
mismatches too hastily when comparing profiles.
In kinship analysis, a minimum of 12 independent
markers is recommended [7]. Nine independent loci are not
considered discriminatory enough in a DVI situation [20].
There are a number of suitable kits on the market which
include the Codis, European and Interpol standard (ESS)
sets of markers (Table 2).
SNPs
Single Nucleotide Polymorphisms (SNPs) are single base
substitutes at certain positions on the DNA molecule. As
the DNA fragments are very short, they are useful for
degraded samples. There are millions of defined SNPs in
the human genome but to be useful for a DVI-situation and
kinship analysis, they need to be well separated. Kinship
analyses indicate that about 5,000 SNPs is the upper limit
to reveal any kinship, above this number, linkage disequilibrium complicates the calculations significantly
favoring the genetically closest relationship [21]. SNP
markers can be used in informative clusters, an approach
that provides a possible solution to the problem with
Table 2 STR-markers distributed into three common sets used for
DNA analyses for identification and forensic purposes
Marker

ESS-loci

CODIS-loci

D3S1358

ESS ? CODIS-loci
X

D8S1179

D18S51

D21S11

FGA

TH01

vWA

D5S818

D7S820

D13S317

D16S539

CSF1PO

TPOX

D1S1656

D2S441
D2S1338

X
X

D10S1248

D19S433

D22S1045

D12S391

linkage. One STR marker is equivalent to about fifteen

SNPs and a useful set of 52 SNPs for kinship analysis has
been developed and tested [22]. A disadvantage is that SNP
analyses on array platforms need amounts of DNA that
might be difficult to obtain from degraded material. SNPs
were successfully used to identify victims in the 9/11
World Trade Center attack [23].
mtDNA
Mitochondrial DNA (mtDNA) are small circular, DNAmolecules inherited on the maternal line. Their circular
shape makes them more resistant to degradation and they
are more abundant than the single copy STR loci [24].
However, they are not as informative as nuclear autosomal
STR markers. In a closed accident where victims are
expected to be unrelated, the mtDNA might serve the
purpose very well.
Y-chromosome DNA
There are both STR and SNP markers on the Y-chromosome.
These markers are useful when establishing paternal lineages
[25]. One example is the identification of three Swedish boys
in a co-cart team, who were missing, together with their
fathers, in an air plane crash in Milano 2001. The fathers
were identified with methods other than DNA analysis. PM
samples collected from the identified fathers, provided AM
data to identify the sons using Y-chromosome analysis.
Y-chromosome STR markers were also used in the identification of the crew on the Swedish DC3 that was shot down
during a mission in 1952. The aircraft was rescued in 2004
after more than 50 years in the Baltic Sea [26].

Software and statistics

In any DVI-situation conducted by Interpol, DVI-System
International [27] is the software used to assemble all the
data collected. This includes dental information, physical
details, and information on personal belongings, as well as
AM and PM DNA profiles together with sample information. However, additional software is needed to process the
DNA data. Ideally, profiles are to be compared, calculations automated, inconsistencies and partial profiles considered and, a report generated with a conclusion. Although
STR analysis is the first choice of markers used, the software needs to be able to handle Y-chromosome, mtDNA
and SNP data. In addition to the software required for the
matching and reporting, the laboratory performing the
analyses should also have a Laboratory Information Management System (LIMS) to track the samples through the
analysis.

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In recent years, several software systems intended for

DNA DVI data have been launched [2831] and some have
been used in DVI operations (WTC 2001, air plane accident outside Brazil 2009, Australian Bushfires 2009).
Drabek has looked at fifteen different pieces of software
intended for kinship analysis, and validated two of them
[32], Paternity Index [33] and Familias [34].
Software needs to be able to handle three parts of the
DNA process: (1) Registration of data, (2) Laboratory
routines, (3) Matching, calculation and reporting.
It is not necessary for one piece of software to undertake
all three components, however, the software should be
validated at the investigating laboratory. Software should
be characterized by simplicity; be easy to learn and be user
friendly. In a DVI situation, there is no time to focus on
complex software solutions. Also worthwhile is the softwares ability to communicate with DVI System International, as well as software connected to specific analyzing
instruments. A suggestion could be a DNA module especially designed to be compatible to DVI-System International (Fig. 1).
A match of AM and PM DNA data can either be a
kinship match or a direct match, but either way, it is recommended that the likelihood ratio (LR) of the match is
calculated to indicate the strength of the evidence. This
enables the DNA data to be combined with probabilities
from other disciplines for identification in a Bayesian
analysis of the total evidence [7]. The prior probability is
unique for each disaster and depends on many factors
including the number of victims and the geography of the
disaster area. This prior probability in combination with the
LR determines the posterior probability. The prior probability may change as the identification process unfolds.
There has been some discussion on the necessity of using
ethnic population databases as references when performing
DNA analysis. One solution has been to use several
appropriate population databases for each calculated
match. However, if AM data from two first-degree relatives
is collected, or equivalent, the importance of the origin of
the population database is limited. Mathematical questions

DNA AM
Information
from Typing
Laboratory

DNA Information
Transfer moduls

DVI SYSTEM
International
PLASS DATA

DNA Matching
module

DNA PM
Information
from Typing
Laboratory

Fig. 1 Example of DNA analysis software modules communicating

with DVI International System

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that might follow a DNA identification process in a mass

disaster have been discussed with a conclusion implying
that the perfect calculation is rather complicated, or even
not possible, but that relevance to the practical problem is
necessary [35, 36]. Simulated identifications have been
reported, where 13 CODIS STR markers were used. Calculations were performed and ranked according to the
information content of different combinations of relatives
[37]. This list can serve as a guide when determining from
which relatives to request a sample. It is recommended that
15 autosomal STR markers are used to increase the LRs
[7]. In the 2009 Victorian bushfire disaster, Profiler Plus
(nine autosomal markers) (Applied Biosystems) was used
and was not sufficiently discriminatory [38]. They found
several cases where the strength of the match was not
sufficiently indicative, and also a PM profile that could be
linked to two missing persons.
During a DVI operation in a small, isolated community
there might be difficulty distinguishing victims due to a
high degree of relatedness in the population [6]. Another
potential problem is the possibility of false inclusions in
large, open disasters if only one reference AM profile is
available for comparison. In both of the cases described,
two first degree relatives will minimize the number of false
inclusions [3941]. One example from the identification
process after the South East Asian tsunami was a Swedish
mother, who matched to a victim in one allele at all 15
Identifiler (Applied Biosystems) markers. However, the
probability of the match being correct was below 95%, and
it was put aside for further testing. Later, a bio bank sample
(PKU-sample) from the alleged victim was tested. The
profile derived from the bio bank sample matched the AM
profile from the mother as expected, but showed a different
DNA profile from that of the victim PM sample profile,
highlighting the difficulties of having only one first degree
relative in a large disaster. The question of inbreeding as a
risk for false inclusions has also been discussed. Kracun
et al. [42] has shown in an in silico model that this
concern is exaggerated. They found that in a population of
100 persons, ten generations were not enough to generate
visible inbreeding. Inbreeding is indicated by the deviation
from HardyWeinberg equilibrium.

Quality
Quality assurance protocols must be adhered during the
whole identification process. In the potentially chaotic
environment following a disaster, the DVI work must be well
coordinated. DNA samples must be handled taking chain of
custody requirements into account, and processed with validated methods and markers by accredited labs. The practice
of testing duplicate aliquots of a sample is an established

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method used to reveal any accidental mix-ups when handling

numerous samples. Personnel should be properly trained and
used to DNA analysis and evaluation. Byard and Winskog
describe a scenario with a DVI-industry where mobilized
teams may not be engaged in current professional practices
[43]. In such circumstances, the risk for evoked mistakes is
obvious. The authors also suggest a quality standard Correction of Failure Index (CF); the total number of cases/
specimens that has to undergo re-examination as a percentage of the total number of cases/specimens examined [44].
This could apply both to samples and bodies. CF should
clearly be as close to zero as possible.
It is also expected that information will be handled in a
secure way, and data should be exchanged in a coded
manner. There is specialized software able to send data in a
coded way from sites that lack secure network connections.

The psychological health among DVI personnel has also

been discussed. In Hurricane Katrina, geneticists from around
the country volunteered for the DNA identification efforts in
the fall 2007. A survey showed that a vast majority volunteered because they felt their professional skills as geneticists
were needed and they would like to see the genetic community
more actively involved in mass fatality response. One hundred
percent of the respondents said they would volunteer again
[47]. Compared to professional DVI staff workers, other
volunteers tend to have higher complaint levels. Factors that
contributed to mental health complaints of volunteers included identification with victims as a friend, severity of exposure
to gruesome events during disaster work, anxiety sensitivity,
and lack of post social support [48]. It is beneficial for any staff
member if debriefing or other post disaster social support is a
part of the DVI organization.

Health concerns among personnel

Exercises as a part of preparation

In April 2005, Morbidity and Mortality Weekly Report

(MMWR) published a number of recommendations
regarding health concerns for people associated with the
DVI operation in Thailand [45]. After interviewing twenty
DVI-workers at Wat Yan Yao, new guidelines were
adopted, e.g. separate areas for food and refreshments.
Some members of the staff expressed concerns regarding
contracting infections from the bodies. A temporary
occupational health clinic provided tetanus vaccinations for
staff members at the morgue. The same concerns may be
expressed among laboratory personnel handling PM samples and adequate vaccinations should be offered.
The catastrophe in Japan in March 2011 highlighted that
the disaster area itself could pose a potential health risk to
the personnel working in the area. The Interpol Disaster
Victim Identification guide dedicates a chapter on how to
deal with chemical, biological or nuclear substances
(CBRN) [1]. Even if the DNA samples are only a tiny part
of a potentially contaminated body, the laboratories should
be prepared to receive material contaminated with radioactivity, hazardous chemicals, or biohazards.
There is an ongoing discussion about the risk of contaminants through bioterrorism, where disease-causing
organisms are used against civilian populations. In such a
case the DVI personnel need to be protected from contagious pathogens. It has been reported that up to 50 kGy
gamma irradiation successfully neutralizes pathogens
without damaging the STR profile of the sample. Some
degradation in peak heights was noted, but a full ninemarker profile was obtained where the radiation dose was
between 10 and 50 kGy [46]. Of course, specific equipment
and facilities are needed to protect the personnel from the
gamma irradiation.

It is difficult to prepare for an event that strikes unexpectedly. One way is to organize exercises to practice
different approaches in the DVI operation, such as team
integration, organization and behavioral responses. The
outcome of a DVI-exercise, Operation Torch is described by Rutty and Rutty [49]. They found that the organizers and the participants had different objectives for the
exercise. The organizers were focused on where there were
gaps in the capabilities, while the participants objectives
were to learn more, and they saw Operation Torch as an
educational event. The authors question the effectiveness
of an exercise if not everyone has the same intention and
suggest a national center with a core evaluation strategy.

Conclusion
Irrespective of whether there is an accident with a single
victim or a mass catastrophe with numerous victims, identification of the deceased is an important part of the overall
response. Cooperation between different professions and
disciplines is crucial, especially if the catastrophe involves
several nationalities. DNA identification is one discipline
that is able to help relatives repatriate their loved ones and to
proceed with the legal aspects concerning the deceased.
Key points
The process to identify a victim by DNA includes:
1.
2.

collection of best possible AM samples

the choice of PM samples in the best condition
possible

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3.

4.
5.

Forensic Sci Med Pathol (2012) 8:140147

DNA analysis, matching of AM and PM data, and

statistical weighting of the match. Data is best handled
electronically.
Quality must be prioritized throughout the process.
Cooperation between different disciplines in DVI work
is crucial.

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