Analysis of mitochondrial SNPs in addition to conventional STR-typing in a case of aggravated theft

In the field of forensic DNA typing, the analysis of Short Tandem Repeats (STRs) can fail in cases of degraded DNA. The typing of coding region Single Nucleotide Polymorphisms (SNPs) of the mitochondrial genome provides an approach to acquire additional information. In the examined case of aggravated theft, both suspects could be excluded of having left the analyzed hair on the crime scene by SNP typing. This conclusion was not possible subsequent to STR typing. SNP typing of the trace on the torch light left on the crime scene increased the likelihood for suspect no. 2 to be the origin of this trace. This finding was already indicated by STR analysis. Suspect no. 1 was excluded for being the origin of this trace by SNP typing which was also indicated by STR analysis. A limiting factor for the analysis of SNPs is the maternal inheritance of mitochondrial DNA. Individualisation is not possible. In conclusion, it can be said that in the case of traces which cause problems with conventional STR typing the supplementary analysis of coding region SNPs from the mitochondrial genome is very reasonable and greatly contributes to the refinement of analysis methods in the field of forensic genetics.     

Identification of the source of ivory idol by DNA analysis

In this study, we describe a forensic case dealing with the identification of the source of the processed ivory object by DNA analysis. Two pieces of Lord Krishna's idols from a shop were confiscated by an investigating agency of the Indian government and forwarded to us to identify the source of its origin. We succeeded in isolating DNA from both processed ivory idols by using the phenol/chloroform DNA extraction method. The extracted DNA was subjected to PCR amplification using an elephant-specific mitochondrial DNA (mtDNA) D-loop marker. DNA sequence analysis of the amplified fragment of mtDNA D-loop region confirmed that the idols were consistent with Asian elephant with 99% similarity.     

Using hydrophilic adhesive tape for collection of evidence for forensic DNA analysis

Known exemplar samples of human DNA have traditionally been body fluids, such as blood, saliva, and semen. In each case, the presence of water is a risk for the bacterial growth, which may degrade the DNA evidence. In this study, the authors have developed a method that employed a hydrophilic adhesive tape (HAT) for collecting DNA evidence. The HAT method was used to remove surface cells from relatively hairless areas on the body. The area examined were ankle, arm, behind the ear, between fingers and back of the neck. The HAT was then dissolved in the extraction buffer. DNA typing was performed at vWA, THo1, F13A1, and FES loci using the short tandem repeat (STR) analysis. Our results show that the samples collected from ear give the best results with a success rate of 100%. All subjects tested by this method had known STR genotypes established from buccal swabs. The authors' results suggest that the HAT method can be used as a less invasive method for collecting biological evidence for forensic DNA analysis. In addition, this collection method should reduce the risk of DNA degradation due to the moisture, which is encountered using conventional collecting methods.     

Identification of Human Remains by DNA Analysis of the Gastrointestinal Contents of Fly Larvae

Dipterous fly larvae (maggots) are frequently collected from a corpse during a criminal investigation. Previous studies showed that DNA analysis of the gastrointestinal contents of maggots might be used to reveal the identity of a victim. However, this approach has not been used to date in legal investigations, and thus its practical usefulness is unknown. A badly burned body was discovered with its face and neck colonized by fly larvae. Given the condition of the body, identification was not possible. Short tandem repeat (STR) typing was performed using the gastrointestinal contents of maggots collected from the victim and was compared to STR profiles obtained from the alleged father. The probability of paternity was 99.685%. Thus, this comparative DNA test enabled the conclusive identification of the remains. This is the first reported case of analysis of human DNA isolated from the gastrointestinal tract of maggots used to identify a victim in a criminal case.     

Combined Statistical Analyses of Forensic Evidence in Sexual Assault: A Case Report and Brief Review of the Literature

DNA analysis has been widely used in the forensic field in order to contribute to identifying the perpetrator of a crime. Forensic investigation in sexual assaults usually focuses on locating and identifying biological fluids, followed by DNA analysis. The identification of certain compounds present in condoms can be useful to reconstruct the occurred event, especially in cases of sexual assaults where the DNA analysis did not show the presence of a male profile and where RNA analysis did not show the presence of sperm markers. Herein we describe the case of a woman reporting to be victim of sexual assault, who was not able to provide accurate information concerning the dynamics of the event; she remembered only forced penile–vaginal penetration by a single perpetrator. We performed short tandem repeat (STR) analyses and mRNA typing for forensic genetics testing on vaginal and rectal swabs collected on the victim, and Fourier-transform infrared spectroscopy (FTIR) followed by chromatographic analyses for the detection of condom compounds on the same swabs. The STR analysis showed only the victim’s genetic profile, and RNA analysis showed only the presence of vaginal and skin markers. In this situation, the identification of condom compounds residues on vaginal swabs became important as it complemented other collected evidences allowing the Court to reconstruct the events. A proposal of likelihood ratio (LR) calculation for the assessment of the weight of evidence in this case is described.     

False‐Positive Results with Amylase Testing of Citrus Fruits

In a case of robbery in which the criminals passed through the garden adorned with calamondin trees (Citrus madurensis), the investigators found in the grass six calamondin fruits, some undamaged, while others apparently bitten. The fruits were collected and sent to the laboratory for DNA analysis to verify the presence of saliva and robbers' DNA profile. A specific immunochromatographic strip test for saliva confirmed the presence of human salivary α‐amylase, but similar positive results were also observed for intact calamondin and other citrus fruits. Further analysis with a specific automated amylase test confirmed the absence of amylase activity. DNA quantification and typing using a specific forensic kit revealed no human DNA presence in any fruits. This case report demonstrates for the first time the occurrence of false positives when human saliva is sought on citrus fruits.     

Analysis of Mitochondrial DNA from a Burned,Ninhydrin‐Treated Paper Towel

Contact‐based evidence is likely to have limited quantities of DNA and may yield mixed profiles due to preexisting or contaminating DNA. In a recent arson investigation, a paper towel was collected and used as circumstantial evidence. The paper towel was partially burned and was likely set on fire with flammable liquid. As part of the investigation, the paper towel was treated with ninhydrin to visualize fingerprint evidence. Initial DNA analysis of two swabs was negative for short tandem repeat (STR) markers and revealed a mixture of mitochondrial DNA (mtDNA). Analysis of 13 additional cuttings yielded four more mixed profiles, but also two samples with a common single‐source profile. The single‐source mtDNA profile matched that of the primary suspect in the case. Thus, even if initial mtDNA analysis yields a mixed profile, a sampling strategy involving multiple locations can improve the chance of obtaining valuable single‐source mtDNA profiles from compromised evidence in criminal casework.     

DNA Recovery From Tape-Lifting Kits: Methodology and Practice

In the case of suspicious deaths, the technique of 1:1 taping is often used in Belgium. It consists of affixing a large number of adhesive tapes to the body of the victim. It is conventionally aimed at obtaining microtraces (e.g., fibers, hair) and is usually not used for DNA analysis. However, in some cases, DNA analysis of certain areas of interest identified on the 1:1 taping material can offer a last resort solution. The four-step method that is described in this article involves the selection of areas of interest on the body (Step 1), the selection of the corresponding tapes (Step 2), the decontamination of the tapes (Step 3), the selection of areas of interest on the tapes, for DNA sampling (Step 4). The method is illustrated by its successful application in four murder cases. In each case, DNA profiles of good quality could be identified, including profiles of persons different from the victim.     

Paternity testing: blood group systems and DNA analysis by variable number of tandem repeat markers

Two recent paternity cases are reported. In the first case of paternity exclusion, deoxyribonucleic acid (DNA) restriction fragment length polymorphisms (RFLPs) on variable number of tandem repeat (VNTR) loci with multiple alleles were informative, as well as established systems of red blood antigens, red cell enzymes, serum proteins, and human leukocyte antigens. In the second case, in which both the alleged father and the first wife were deceased, the paternal genotype was determined by using genetic markers from the second wife and four children, which then were compared with the paternal alleles of the child in question, the plaintiff in this case. The high probability of paternity (0.999,998,7) made us conclude that the man probably was the actual father. The DNA analysis by VNTR probes appears to be quite valuable in the study of paternity cases.     

STR genotyping and mtDNA sequencing of latent fingerprint on paper

A systematic study was conducted to investigate whether DNA can be successfully extracted from latent fingerprints deposited on ordinary paper and analysed using short tandem repeat profiling and mitochondrial DNA sequencing. In order to evaluate the performance of latent fingerprint analysis in a criminal case, experiments with varying conditions were carried out to improve our understanding of low copy number (LCN) DNA typing. After optimising the extraction methods to achieve increased sensitivity, the examination of touched paper can routinely yield the STR profile of the individual who has touched it. A fingerprint can therefore be considered as a potential source of DNA for genetic identification. Nevertheless, the findings of our "after enhancement experiment" (using chemically or physically pre-treated fingerprints), and our "mixture experiment" (using fingerprints from three to four people on the same sheet of paper) help to define the limitations of the low copy number PCR technique in forensic casework.     

IGNA's original LIMS: A complete traceability of administrative and analytic processes for forensic cases

IGNA is the first French laboratory for forensic DNA analysis. IGNA LIMS was developed from the software Microsoft Dynamics NAV which is an ERP (Enterprise Resource Planning) with a real opened solution for specific development and compatible with a lot of Microsoft applications.Thanks to the LIMS, we assure the traceability of analysis and also administrative process (traceability of letters, quotes or phone calls for each case and registration of actions to be done in a case (call for results …)).For one penal case, we can associate one or more sealed items. The DNA expert indicates the analysis to do on each piece (biological fluid detection, type of extraction, nuclear or mitochondrial DNA analysis …).During the technical process, samples automatically pass from a step to the next. Technicians indicate all the information needed for traceability (consumables lot number, used robots …).At the end of the process, the DNA expert validates his results, which are then directly transferred to the final report.Development of the IGNA LIMS from Microsoft Dynamics NAV allows us to use all specific applications of an ERP such as purchases, marketing, sales, etc. and to assure computerization of all our process, from quote to analysis report.     

Decision support for using mobile Rapid DNA analysis at the crime scene

Mobile Rapid DNA technology is close to being incorporated into crime scene investigations, with the potential to identify a perpetrator within hours. However, the use of these techniques entails the risk of losing the sample and potential evidence, because the device not only consumes the inserted sample, it is also is less sensitive than traditional technologies used in forensic laboratories. Scene of Crime Officers (SoCOs) therefore will face a ‘time/success rate trade-off’ issue when making a decision to apply this technology.In this study we designed and experimentally tested a Decision Support System (DSS) for the use of Rapid DNA technologies based on Rational Decision Theory (RDT). In a vignette study, where SoCOs had to decide on the use of a Rapid DNA analysis device, participating SoCOs were assigned to either the control group (making decisions under standard conditions), the Success Rate (SR) group (making decisions with additional information on DNA success rates of traces), or the DSS group (making decisions supported by introduction to RDT, including information on DNA success rates of traces).This study provides positive evidence that a systematic approach for decision-making on using Rapid DNA analysis assists SoCOs in the decision to use the rapid device. The results demonstrated that participants using a DSS made different and more transparent decisions on the use of Rapid DNA analysis when different case characteristics were explicitly considered. In the DSS group the decision to apply Rapid DNA analysis was influenced by the factors “time pressure” and “trace characteristics” like DNA success rates. In the SR group, the decisions depended solely on the trace characteristics and in the control group the decisions did not show any systematic differences on crime type or trace characteristic.Guiding complex decisions on the use of Rapid DNA analyses with a DSS could be an important step towards the use of these devices at the crime scene.     

Challenging DNA: Assessment of a range of genotyping approaches for highly degraded forensic samples

It is common in forensic casework to encounter highly degraded DNA samples from a variety of sources. In this category bone and teeth samples are often the principal source of evidential material for criminal investigations or identification of long-deceased individuals. In these circumstances standard STRs are prone to fail due to their long amplicon sizes (since DNA becomes progressively more fragmented as it degrades). To successfully resolve such cases alternative markers can be used and until recently the only other tool available was mitochondrial DNA, which despite being more resistant to degradation, is much less informative. A rapidly developing approach to analyzing degraded DNA is the typing of loci from short-amplicon PCR products based on markers such as mini-STRs and autosomal SNPs. We have performed an analysis of several cases with naturally degraded DNA using established STRs plus mini-STRs and autosomal SNPs in order to make an objective comparison of the performance of each method using challenging DNA. The main aim was to establish the benefits and drawbacks of each marker set to help the practitioner choose the DNA analysis method most suited to the circumstances of each case.     

Detection of fetal DNA in a cell pellet after centrifugation of mountant

In order to obtain fetal cells (e.g., for paternity cases) after abortion, the centrifugation of mountant (in our case formalin) may be tried when the DNA examination of the fixed tissue itself gives limited or no (or not enough) information. The fixed tissue was microscopically negative for fetal cells and gave no satisfactory results when examined for DNA. Centrifugation of approximately 50 mL of reddish colored formalin resulted in a cell pellet that was examined for DNA, which gave enough information to confirm a case of sexual abuse.     

Touch DNA of Shed Skin Cells from the Deployed Airbag to Address Drunken Driving Crimes

In the criminal cases of driving under the influence （DUI）, DNA evidence can be collectedfrom the deployed airbag of the motor vehicle and submitted to the crime lab for touch DNA analysis.The evidence can be acquired when the skin cells are observed on the surface of the airbag in a trafficaccident. However, the low quantity or quality of the evidence collected from a crime scene preventsfurther identification analysis in many cases. In the current study, we reported a case of identifyingtouch DNA extraction from the shed skin cells from the deployed airbag of a motor vehicle. We man-aged to collect DNA evidence from the shed skin cells in an airbag using a proper approach of collec-tion and extraction. The 5.87 ng of extracted DNA was sufficient for genotyping and forensic identifica-tion, which helped to identify the driver of the car in collision with a pier in the street. In DUI casesand other traffic accidents, therefore, the amount of touch DNA extracted from the deployed airbag canbe sufficient for DNA marker genotyping and further analysis.     

The value of skull-photo superimpositions in identifying corpses found in domestic settings

The identification of decomposed corpses found in domestic settings is frequently problematic because comparative material for methods such as forensic odontostomatology, comparative X-ray analysis, or DNA analysis, is not available. In the case presented here, a photograph from an old, expired passport could be used to successfully identify a "domestic setting" corpse in a skull-photo superimposition. In an additional DNA analysis, 13 STR-loci could be amplified from tissue samples taken from the corpse. DNA comparison with the presumed brother of the deceased yielded a probability of 97.09% for siblingship. Y-STR-analysis was, therefore, performed. The results showed that all of the systems for the presumed brother and the corpse conformed, with the exception of the DYS390 locus, in which allele 21 was found for the corpse and allele 22 for the brother. Despite the rapid development of other identification procedures, skull-photo superimpositions remain an important means of identification. Last not least this is due to the increasing ubiquity of personal photo documents in the age of digital photography. The validity of the results from a DNA analysis in an identification process depends largely on the authenticity of the samples available for comparison and the degree to which the DNA from the corpse is preserved. In the case presented by the authors, positive identification of the corpse solely on the basis of the DNA analysis would not have been possible. Numerous constellations can be imagined for decomposed corpses found in domestic settings for which skull-photo superimpositions may be the only possible option for identifying the corpse.     

Identification of a forensic case using microscopy and forensically informative nucleotide sequencing (FINS): A case study of small Indian civet (Viverricula indica)

The exhibits obtained in wildlife offence cases quite often present a challenging situation for the forensic expert. The selection of proper approach for analysis is vital for a successful analysis. A generalised forensic analysis approach should proceed from the use of non-destructive techniques (morphological and microscopic examination) to partially destructive and finally destructive techniques (DNA analysis). The findings of non-destructive techniques may sometime be inconclusive but they definitely help in steering further forensic analysis in a proper direction. We describe a recent case where a very small dried skin piece (< 0.05 mg) with just one small trimmed guard hair (0.4 cm) on it was received for species identification. The single guard hair was examined microscopically to get an indication of the type of species. We also describe the extraction procedure with a lower amount of sample, using an automated extraction method (Qiagen Biorobot EZ1^®) and PCR amplification of three mitochondrial genes (16s rRNA, 12s rRNA and cytochrome b) for species identification. Microscopic examination of the single hair indicated a viverrid species but the initial DNA analysis with 16s rRNA (through NCBI BLAST) showed the highest homology (93%) with a hyaenid species (Hyaena hyaena). However, further DNA analysis based on 12s rRNA and cytochrome b gene proved that the species was indeed a viverrid i.e. Viverricula indica (small Indian civet). The highest homology shown with a Hyaenid species by the 16s rRNA sequence from the case sample was due to lack of a 16s rRNA sequence for Viverricula indica in the NCBI data base. The case highlights the importance of morphological and microscopic examinations in wildlife offence cases. With respect to DNA extraction technology we found that automatic extraction method of Biorobot EZ1^® (Qiagen) is quite useful with less amount of sample (much below recommended amount).     

'The story of Abraham,Isaac and Jacob" or "am I my brother's keeper?"

Presented is a case report of a violent sexual assault where the DNA profile obtained from an item of evidence was compared to a suspect's profile. The profiles did not match, but the sharing of such a large number of alleles raised the suspicion that perhaps the real perpetrator was a blood relative of the suspect. The investigators requested a sample from the suspect's brother, and a match was defined. In an era of technological breakthroughs in the field of forensic DNA analysis, the importance of the scientist's attention to the evidence presented in each case is stressed.     

降解生物检材DNA分析研究新进展

在法医检案过程中,如何从降解生物检材中获得正确的DNA分型是法医学界的一个难题。本文对降解生物检材DNA分析研究取得的新进展进行综述,从检案的各个环节出发对近年来提高DNA分型成功率的方法和技术进行了详细介绍,例如新型遗传标记的开发和二代测序技术的应用等。希望为降解检材的分析提供新的思考和方法。     

Recovery of trace DNA and its application to DNA profiling of shoe insoles

In recent years, the analysis of trace amounts of DNA has become a necessary and useful forensic tool. DNA profiles can be obtained from items that have been worn or handled, due to the presence of transferred DNA derived from skin cells. Shoeprints collected from crime scenes that match a suspects shoe can link a shoe to the crime scene. A DNA profile from inside the shoe can link a wearer to a shoe thus increasing the evidential value of the forensic evidence. In this work, variation in the amount of DNA recovered from hands and feet of different individuals is investigated. Sites for sampling DNA from shoe insoles are compared and a protocol for the subsequent sampling and extraction is developed. Finally, a case study is described where DNA analysis of shoe insoles has provided forensic evidence.