Vous êtes sur la page 1sur 3

J Indian Acad Forensic Med, 32(4) ISSN 0971-0973

Original research paper

Application of Victims‟ Fingernails in Forensic DNA Analysis


*Kamoun Arwa, *Mahfoudh Nadia, **Ayadi Adnene, **Hammemi Zouheir, **Maatoug Samir,
**Makni Hafedh

Abstract
DNA extracted from the victims‟ fingernails may assist in the identification of the aggressor. Fingernails
were collected from 8 victims, and were subjected to DNA extraction using the Kit « Tissue and Hair Extraction Kit
(Promega) ».
All samples were typed for 15 autosomal short tandem repeats and for amelogenin using the Kit
« Powerplex TM16 system (Promega) » and the ABI Prism 310 DNA sequencer. The profiles obtained were
compared with those achieved by similar typing of victims‟ and suspects‟ blood.
In two Forensic investigations, mixed genotypes were detected in DNA extracted from the nails: Alleles originating
from the victim were coamplified with other alleles that matched the suspect‟s genotypic profile. This indicated that
victims‟ fingernails contained biological material (blood, epithelial cells) originating from the suspect.
Our results confirmed the usefulness of the nails as a specimen for forensic identification of the aggressor.

Keywords: Fingernails, DNA, Victim, STR, Promega, Genotypic Profile, Fingernails, Hair, Tissue

Introduction: Blood collected from the 10 victims and


DNA analysis has proven to be a valuable from 13 suspects, and bloodstains sampled from the
technique for human identification and for the crime scenes. Samples were stored at -20°C until
resolution of criminal disputes.Human nail material DNA extraction.
has been identified as a potential source of biological DNA extraction from blood was carried out
material for Forensic DNA testing [1, 2, 3]. by salting out technique.
Fingernail clippings collected from victims Bloodstains and fingernails were extracted with DNA
in assault cases, principally sexual cases, are IQTM system according to the protocol described by
occasionally sent to Forensic laboratories as a Promega [5]:
possible source of DNA originating from the For bloodstains, 150µl of lysis buffer was
perpetrator [4]. added and incubated for 30 mn at 70°C. Lysis buffer
The aim of the present study was to optimize and sample were then transferred to a DNA IQ Spin
the extraction conditions using the Kit DNA IQTM Basket seated in a 1.5ml microcentrifuge tube and
system (Promega) in order to identify a foreign Centrifuged at room temperature for 2 minutes at
profile in victims‟ fingernails possibly originating maximum speed.
from the perpetrator. DNA was purified by adding 7µl resin. After washing
Materials and Methods: and drying of the resin, DNA was eluted in 50µl of
elution buffer. The nail clippings received no
Extraction of DNA treatment before extraction.
Within 8 forensic caseworks, we received DNA extraction from nail clippings was
the fingernail clipping of 10 victims of murder (7 carried out by a modification of the protocol
males and 3 females). described above. In order to minimize the quantity of
Corresponding Author: endogenous DNA recovered, digestion was done at
*Dr Kamoun Arwa room temperature and not at 70°C for a short period
Hôpital Hédi Chaker, Route El Ain Km 0.5, 3029 of time ranging from 1 minute to 10 minutes, mixing
Sfax, Tunisia was done by a gentle pipetting and not by vortexing.
E-mail : arwakamoun@yahoo.fr DNA purification and elution steps were not
Tel: 216 98652824, Fax: 216 74248622 modified.
** Professor Short Tandem Repeat Amplification And
Service de médecine légale, Typing:
EPS Habib Bourguiba,
The DNA samples (1ng) were amplified
3029, Sfax, Tunisia
using the powerplex 16 Kit following the
manufacturer‟s recommendations. The amplified

289
J Indian Acad Forensic Med, 32(4) ISSN 0971-0973

products were analyzed using an ABI Prism 310 When studying the resolving power of the
Genetic Analyzer according to the manufacturer‟s powerplex 16TM system, Krenke at al found that only
recommended protocol. 17% of minor alleles could be detected at a ratio of
Initial fragment sizing was performed by the 1:19. [7]
GeneScan Software (Applied Biosystems). Allele Gangitano et al optimized a non organic DNA
calling was performed by Promega‟s PowerTyper TM extraction procedure for fingernail clippings after
16 macro operating within the genotyper® software scratching. [8]
program (Applied Biosystems). DNA samples were typed for an STR locus
Genetic profiles obtained from the nail clippings residing on the Y chromosome: DYS19. The success
were compared to those typed from blood samples. rate of typing of the scratched person was 64%. This
Results: strategy based on the identification of haplotypic
In the 1st Forensic casework, the victim was markers could be useful only for the exclusion of
a man. The nail clippings digestion was carried out at male suspects.
room temperature for 10 mn. Genetic profile analysis Wiegand et al reported that a foreign profile
revealed evidence for the presence of DNA from the could be obtained from debris scraped from
offender in fingernail clippings of the victim‟s hands, underneath nails if removal of particles was carried
with allele signal intensities 3 times lower than those out with sufficient care. [9]
of the victim [figure 1]. Cline et al developed a technique for
In the second investigation, the victim was a isolating and purifying foreign DNA in fingernail
woman. Victim‟s nails were soaked in the lysis buffer clippings [10]: human test nails were heavily coated
for only 2 mn at room temperature. Amplification with mouse liver and allowed to dry several days. A
with the powerplex 16TM system showed a foreign one hour H2O/EDTA (ethylenediaminetetraacetic
male genetic profile which could be assigned to the acid) soak of contaminated nail clippings released
perpetrator [figure 2]. only exogenous DNA. The presence or absence of
In the other caseworks, fingernails collected each species DNA was confirmed through
from 8 victims were digested for only 1mn at room mitochondrial DNA amplification using PCR sites
temperature. Genetic profiles typed were identical to conserved in all mammals.
those obtained from the blood nail donors. To date, genetic identification of foreign
In one investigation, genetic profiles retrieved from 2 DNA in fingernail clippings was successful in 2
bloodstains matched the profile of one suspect, cases. In the 1st report, debris from the fingernails of
providing hence evidence for his culpability [figure the suspect were scraped out with a plastic spatula
3]. and extracted with Chelex 100. Amplification with
Discussion: the pentaplex kit genRES MPX revealed alleles at all
DNA Identification is often useful in loci which could be assigned to the victim. [11]
forensic investigation, since it could originate from In the 2nd case, a 2 years old micro-
the perpetrator, particularly when the sample is taken bloodstain under the fingernails of a victim was
from the victim cadaver. Indeed, sexual assaults or scraped. A mixed DNA sample from both the victim
homicides are often associated with multiple actions and the scraped person was recovered. [9]
of aggression and defence which may lead to transfer In our report, we extracted DNA from the
of DNA containing material: skin epithelial cells, entire fingernail clippings of 10 victims by DNA
blood. Therefore, in relevant cases, the analysis is IQTM system. We used mild digestion conditions
focused on fingernail clippings or on debris scraped (shorter incubation time, room temperature, gentle
from underneath nails. [4] Several experimental mixing) in order to minimize victim‟s epithelial cells‟
studies aimed to develop techniques for foreign DNA lysis. We succeeded to identify a foreign DNA
extraction from nails of volunteers having scratched pattern from fingernail clippings in 2 cases. In the
other subjects. other caseworks, failure to identify additional alleles
Oz et al used phenol/chloroform for the could be attributed to the absence of foreign
extraction of DNA from the entire fingernail biological material in the victims‟ fingernails.
clippings. [6] Amplification of 4 autosomal STRs Conclusion:
(short tandem repeats) produced a genetic profile Our study underlines the value of the genetic
identical to that typed from the buccal swabs of the analysis of fingernails in forensic investigations. In
same volunteers. They concluded that the routine fact, victims‟ fingernails may contain biological
fingernail clippings would not contribute essential material which could possibly originate from the
information in forensic casework. In fact, when perpetrator: body fluids scratched epithelial cells.
digesting the entire nails, endogenous DNA would be Digestion conditions must be optimized to minimize
relatively abundant and thus preferentially amplified. extraction of victim‟s DNA.

290
J Indian Acad Forensic Med, 32(4) ISSN 0971-0973

References: 7. Krenke BE, Tereba A, Anderson SJ, Buel E,


Culhane S, Finis CJ, et al. Validation of a 16-locus
1. Tahir MA, Watson N. Typing of DNA HLA-DQ alpha
fluorescent multiplex system. J Forensic Sci. 2002;
alleles extracted from human nail material using
47(4):773-85.
polymerase chain reaction. J Forensic Sci. 1995;
8. Gangitano DA, Garófalo MG, Juvenal GJ, Budowle
40(4):634-6.
B, Padula RA. Typing of the locus DYS19 from DNA
2. Anderson TD, Ross JP, Roby RK, Lee DA, Holland
derived from fingernail clippings using PCR Concert
MM. A validation study for the extraction and analysis
rapid purification system. J Forensic Sci. 2002;
of DNA from human nail material and its application to
47(1):175-7.
forensic casework. J Forensic Sci. 1999; 44(5):1053-6.
9. Wiegand P, Bajanowski T, Brinkmann B. DNA
3. Tahir MA, Balraj E, Luke L, Gilbert T, Hamby JE,
typing of debris from fingernails. Int J Leg Med. 1993;
Amjad M. DNA typing of samples for polymarker,
106:81-83.
DQA1, and nine STR loci from a human body exhumed
10. Cline RE, Laurent NM, Foran DR. The fingernails of
after 27 years. J Forensic Sci. 2000; 45(4):902-7.
Mary Sullivan: developing reliable methods for
4. Keating SM, Allard JE. What's in a name?--Medical
selectively isolating endogenous and exogenous DNA
samples and scientific evidence in sexual assaults. Med
from evidence. J Forensic Sci. 2003; 48(2):328-33.
Sci Law. 1994; 34(3):187-201.
11. Lederer T, Betz P, Seidl S. DNA analysis of fingernail
5. DNA IQ™ System- Small Sample Casework Prototcol.
debris using different multiplex systems: a case report.
Madison (WI): Promega Corporation; 2002.
Int J Legal Med. 2001; 114(4-5):263-6.
6. Oz C, Zamir A. An evaluation of the relevance of
routine DNA typing of fingernail clippings for forensic
casework. J Forensic Sci. 2000; 45(1):158-60.

291