Showing papers in "Journal of Forensic Sciences in 2006"

Genetics and genomics of core short tandem repeat loci used in human identity testing.

Abstract: Over the past decade, the human identity testing community has settled on a set of core short tandem repeat (STR) loci that are widely used for DNA typing applications. A variety of commercial kits enable robust amplification of these core STR loci. A brief history is presented regarding the selection of core autosomal and Y-chromosomal STR markers. The physical location of each STR locus in the human genome is delineated and allele ranges and variants observed in human populations are summarized as are mutation rates observed from parentage testing. Internet resources for additional information on core STR loci are reviewed. Additional topics are also discussed, including potential linkage of STR loci to genetic disease-causing genes, probabilistic predictions of sample ethnicity, and desirable characteristics for additional STR loci that may be added in the future to the current core loci. These core STR loci, which form the basis for DNA databases worldwide, will continue to play an important role in forensic science for many years to come.

Evaluating the Accuracy and Precision of Cranial Morphological Traits for Sex Determination

Abstract: Sex determination is a key analysis that forensic anthropologists perform in order to construct a biological profile of human remains. The techniques used in forensic investigations must meet the Mohan or Daubert criteria, for admissibility in a court of law. In this study, the precision and accuracy of 21 morphological characteristics of the skull were tested on a modern sample of 50 adult crania of European White ancestry. The following craniofacial features are identified as high-quality traits, defined by intraobserver error or=80%: mastoid size, supraorbital ridge size, general size and architecture, rugosity of the zygomatic extension, size and shape of the nasal aperture, and gonial angle. Ninety-six percent accuracy and 92% precision were achieved using 20 traits in combination. Fisher's exact probability tests revealed no significant differences (p=0.05) in the levels of precision or accuracy between age categories. Sex-related bias in accuracy was found for the following cranial features: ramus symphysis (p=0.009), zygomatic extension (p=0.0016), and occipital markings (p=0.0013). These traits demonstrated a greater tendency to be scored male than female.

Development and validation of the AmpFlSTR Yfiler PCR amplification kit: a male specific, single amplification 17 Y-STR multiplex system.

Abstract: In the past 5 years, there has been a substantial increase in the use of Y-short tandem repeat loci (Y-STRs) in forensic laboratories, especially in cases where typing autosomal STRs has met with limited success. The AmpF'STR s Yfiler TM PCR amplification kit simultaneously amplifies 17 Y-STR loci including the loci in the ''European minimal haplotype'' (DYS19, DYS385a/b, DYS389I, DYS389II, DYS390, DYS391, DYS392, and DYS393), the Scientific Working Group on DNA Analysis Methods (SWGDAM) recommended Y-STR loci (DYS438 and DYS439), and the highly polymorphic loci DYS437, DYS448, DYS456, DYS458, Y GATA H4, and DYS635 (formerly known as Y GATA C4). The Yfiler TM kit was validated according to the FBI/National Standards and SWGDAM guidelines. Our results showed that full profiles are attainable with low levels of male DNA (below 125pg) and that under optimized conditions, no detectable cross-reactive products were obtained on human female DNA, bacteria, and commonly encountered animal species. Additionally, we demonstrated the ability to detect male specific profiles in admixed male and female blood samples at a ratio of 1:1000.

The RECON typology of stalking: reliability and validity based upon a large sample of North American stalkers.

Abstract: A new typology of stalking, RECON (relationship and context-based), is proposed, based upon the prior relationship between the pursuer and the victim, and the context in which the stalking occurs. The static typology yields four groups: Intimate, Acquaintance, Public Figure, and Private Stranger. The typology was tested on a large (N = 1005) nonrandom sample of North American stalkers gathered from prosecutorial agencies, a large police department, an entertainment corporation security department, and the authors' files. Interrater reliability for group assignment was 0.95 (ICC). Discriminant validity (p < 0.01) was demonstrated on a variety of demographic, clinical, pursuit, threat, and violence characteristics among and between groups. Findings confirm and extend the work of other researchers, most notably the very high risk of threats and violence among prior sexually intimate stalkers, the very low risk of threats and violence among public figure (celebrity) stalkers, and the negative relationship between stalking violence and psychosis.

Evaluation of Real-Time PCR Amplification Efficiencies to Detect PCR Inhibitors

Abstract: Real-time PCR analysis is a sensitive template DNA quantitation strategy that has recently gained considerable attention in the forensic community. However, the utility of real-time PCR methods extends beyond quantitation and allows for simultaneous evaluation of template DNA extraction quality. This study presents a computational method that allows analysts to identify problematic samples with statistical reliability by comparing the amplification efficiencies of unknown template DNA samples with clean standards. In this study, assays with varying concentrations of tannic acid are used to evaluate and adjust sample-specific amplification efficiency calculation methods in order to optimize their inhibitor detection capabilities. Kinetic outlier detection and prediction boundaries are calculated to identify amplification efficiency outliers. Sample-specific amplification efficiencies calculated over a four-cycle interval starting at the threshold cycle can be used to detect reliably the presence of 0.4 ng of tannic acid in a 25 microL PCR reaction. This approach provides analysts with a precise measure of inhibition severity when template samples are compromised. Early detection of problematic samples allows analysts the opportunity to consider inhibitor mitigation strategies prior to genotype or DNA sequence analysis, thereby facilitating sample processing in high-throughput forensic operations.

The use of Polilight in the detection of seminal fluid, saliva, and bloodstains and comparison with conventional chemical-based screening tests.

Abstract: Biological stains can be difficult to detect at crime scenes or on items recovered from crime scenes. The use of a versatile light source may assist in their detection. The ability of Polilight to locate potential semen, saliva, and blood stains on a range of substrates and at different dilutions was tested. We also tested the use of Polilight in comparison with conventional chemical-based presumptive screening tests such as acid phosphatase (AP), Phadebas, and luminol, often used in casework for detecting potential semen, saliva, and blood stains, respectively. The Polilight was able to locate stains that were not apparent to the naked eye. The color of the material on which a stain is deposited can have an effect on the detectibility of the stain. The Polilight was found to be comparable with the AP and Phadebas tests in terms of its sensitivity. In a comparative study between the AP test and Polilight on 40 casework exhibits, one false-negative result was observed when using the Polilight. On a series of mock casework exhibits it was determined that the Polilight can be used successfully to locate saliva stains for DNA analysis. The sensitivity of luminol for detecting potential bloodstains was greater than that of Polilight; however the Polilight has particular application in instances where a bloodstain may have been concealed with paint. Overall, the Polilight is a relatively safe, simple, noninvasive, and nondestructive technique suitable for use in forensic casework.

Differentiating Writing Inks Using Direct Analysis in Real Time Mass Spectrometry

Abstract: Writing ink analysis is used in establishing document authenticity and the sources and relative ages of written entries. Most analytical methods require removing samples or visibly altering the document. Nondestructive, in situ analysis of writing inks on paper without visible alteration is possible using mass spectrometry with a new ion source called Direct Analysis in Real Time. Forty-three different black and blue ballpoint, black fluid, and black gel inks were examined. Both dyes and persistent but thermally labile components of the inks contribute to the mass spectra, principally as protonated molecules (M+H) + . Numerous ink components were identified from the spectra. The spectra were placed in a searchable library, which was then challenged with two spectra from each of the 43 inks. The best match for each of the challenge spectra was correct for all but one ink, which matched with a very similar ink by the same manufacturer.

Differences among species in compact bone tissue microstructure of mammalian skeleton: use of a discriminant function analysis for species identification.

Abstract: In order to develop an identification key for distinguishing between several mammalian species, bone structure of their compact bone tissue was analyzed using qualitative and quantitative characteristics. Ninety femora of adult male humans, pigs, cows, sheep, rabbits, and rats were studied. The average area, perimeter, minimum, and maximum diameter of 1863 Haversian canals and 1863 secondary osteons were measured using a digital image device. The observed data were first used to evaluate inter- and intraspecies diversity. After that, we applied a discriminant function analysis for differentiation of the species by these variables. Classification functions for investigated species give cross-validated correct classification rates for 76.17% of cases. This percentage value can be increased by integrating conclusions from the qualitative analysis.

Using the acetabulum to estimate age at death of adult males.

Abstract: The acetabular region is often present and adequately preserved in adult human skeletal remains. Close morphological examination of the 242 left male os coxae from the identified collection of Coimbra (Portugal) has enabled the recognition of seven variables that can be used to estimate age at death. This paper describes these variables and argues their appropriateness by analyzing the correlation between these criteria and the age, the intra- and interobserver consistence, and the accuracy in age prediction using Bayesian inference to estimate age of identified specimens. Results show significant close correlation between the acetabular criteria and age, nonsignificant differences in intra- and interobserver test, and 89% accuracy in Bayes prediction. Obtained estimated age of the specimens had similar accuracy in all ages. These results indicate that these seven variables, based on the acetabular area, are potentially useful to estimate age at death for adult specimens.

Sequential monitoring of burials containing large pig cadavers using ground-penetrating radar.

Abstract: Ground-penetrating radar (GPR) was used to monitor 12 pig burials in Florida, each of which contained a small pig cadaver. Six of the cadavers were buried in sand at a depth of 0.50-0.60 m, and the other six were buried in sand at a depth of 1.00-1.10 m to represent deep and shallow burials that are generally encountered in forensic scenarios. Four control excavations with no pig interment were also constructed as blank graves and monitored with GPR. The burials were monitored for durations of either 13 or 21 months, and were then excavated to correlate the decomposition state of the cadaver with the GPR imagery. Overall, this study demonstrated that it may be difficult to detect small cadavers buried in sand soon after they are skeletonized because the area surrounding the body, or the grave, may not provide a strong enough contrasting area to be detected by GPR when compared to that of the surrounding undisturbed soil. Also, depth of burial appears to influence grave detection because bodies that are buried at deeper depths may be detected for a longer period of time due to reduced decomposition rates.

Optimization of DNA extraction from low-yield and degraded samples using the BioRobot EZ1 and BioRobot M48.

Abstract: Robotic extraction of DNA from dilutions of blood and semen using either the BioRobots EZ1 or BioRobots M48 consistently produced lower recoveries than standard organic extractions of the same samples. In an effort to increase the efficiency of robotically extracted DNA, glycogen and carrier RNA were added following cell lysis. The addition of glycogen, postlysis, resulted in no improvement in DNA recovery with the BioRobot EZ1. However, when carrier RNA was added to the cell lysate of limited and degraded samples extracted on the EZ1 or the M48, DNA recoveries dramatically increased four- to 20-fold. DNA yields obtained by robotic extraction in the presence of carrier RNA were as high, or higher, as those obtained by organic extraction lacking carrier RNA, while experiments that utilized carrier RNA in both types of extractions showed increased sensitivity for both methods. Furthermore, carrier RNA substantially increased the recovery of fragmented DNA with the EZ1.

Microchip‐Based Cell Lysis and DNA Extraction from Sperm Cells for Application to Forensic Analysis

Abstract: The current backlog of casework is among the most significant challenges facing crime laboratories at this time. While the development of next-generation microchip-based technology for expedited forensic casework analysis offers one solution to this problem, this will require the adaptation of manual, large-volume, benchtop chemistry to small volume microfluidic devices. Analysis of evidentiary materials from rape kits where semen or sperm cells are commonly found represents a unique set of challenges for on-chip cell lysis and DNA extraction that must be addressed for successful application. The work presented here details the development of a microdevice capable of DNA extraction directly from sperm cells for application to the analysis of sexual assault evidence. A variety of chemical lysing agents are assessed for inclusion in the extraction protocol and a method for DNA purification from sperm cells is described. Suitability of the extracted DNA for short tandem repeat (STR) analysis is assessed and genetic profiles shown. Finally, on-chip cell lysis methods are evaluated, with results from fluorescence visualization of cell rupture and DNA extraction from an integrated cell lysis and purification with subsequent STR amplification presented. A method for on-chip cell lysis and DNA purification is described, with considerations toward inclusion in an integrated microdevice capable of both differential cell sorting and DNA extraction. The results of this work demonstrate the feasibility of incorporating microchip-based cell lysis and DNA extraction into forensic casework analysis.

Sex determination using nonmetric characteristics of the mandible in Koreans.

Abstract: The mandible is the largest and hardest facial bone and retains its shape better than other bones in the forensic and physical anthropologic field. The mandible can be used to distinguish among ethnic groups and between sexes. We examined the morphological characteristics of the mandibles of 102 Koreans of either sex. Of 13 nonmetric items of the mandible, the characteristic that best allowed the sexes to be distinguished was the contour of the lower border of the mandible: rocker-shaped mandibles predominated in males (68.1%), whereas most females (84.6%) exhibited a straight mandible. In addition, the mental region was shaped differently between the sexes: the shape of the chin in most males was generally bilobate or square (91.7%), whereas the chin in females was either square (45.5%) or pointed (54.5%). In this study, the positive predict values of male and female were 92.5% and 73.7%, respectively. Therefore, the nonmetric method used to analyze the mandible in this study can be used for sex discrimination.

Facial soft tissue thicknesses in Australian adult cadavers.

Abstract: Craniofacial identification methods heavily rely on the knowledge of average soft tissue depths. This study measured soft tissue thicknesses of an Australian cadaver sample (N = 33) using published needle puncture techniques at 13 anatomical locations. Data were compared and contrasted with other studies that used essentially identical samples and methods. Full descriptive statistics were calculated for measurements made in this study and means, medians, and modes were reported. Differences between mean values for males and females were found to be minimal (2.2 mm or less) and considerable overlap was found between the groups. There were no statistically significant differences between the soft tissue depths of the sexes (P>0.05). These findings indicate that differences between male and female soft tissue depths are of little practical significance for craniofacial identification and, therefore, data (means, standard deviations, and sample sizes) reported for Australians were pooled across the sexes and the studies. Although these new pooled means have increased statistical power, data distributions at some landmarks were skewed and thus emphasis is placed on median and modes reported for this study rather than upon the collapsed data means.

Relative Degradation of Nuclear and Mitochondrial DNA: An Experimental Approach*

Abstract: Single copy nuclear loci often cannot be amplified from degraded remains, necessitating the analysis of mitochondrial DNA (mtDNA). The success in analyzing mtDNA is generally thought to result from its higher copy number in the cell; however, other factors, such as cellular location or molecular features, may be equally or more important in the superior preservation of mtDNA. To explore and compare mtDNA and nuclear DNA degradation, mouse tissues (muscle, liver, and brain) were allowed to degrade at different temperatures, and the relative degradation of a mitochondrial gene, a single copy nuclear gene, and a multi-copy nuclear gene was assayed using real-time polymerase chain reaction. The tissues were also homogenized, allowing the three loci to degrade in the same cellular environment. Gene copy number and cellular location both influence DNA recovery. In some instances, multi-copy loci could be recovered when the single copy locus could not; however, the pattern of relative DNA degradation changed between whole and homogenized tissues. The overall results indicate that DNA degradation is influenced by multiple factors-including cellular location, chromatin structure, and transcriptional activity-factors that could be used to exploit loci for more robust forensic analysis from degraded biological material.

Laser Microdissection Separation of Pure Spermatozoa from Epithelial Cells for Short Tandem Repeat Analysis

Abstract: Short tandem repeat (STR) analysis is a valuable tool in identifying the source of biological stains, particularly from the investigation of sexual assault crimes. Difficulties in analysis arise primarily in the interpretation of mixed genotypes when cell separation of the sexual assailant's sperm from the victim's cells is incomplete. The forensic community continues to seek improvements in cell separation methods from mixtures for DNA typing. The feasibility of applying laser microdissection (LMD) technology to precisely separate sexual assault cell mixtures by visual inspection coupled with laser dissection was assessed through three experiments. First, various histological stains were evaluated for use with LMD and DNA analysis. Second, different DNA isolation methods were evaluated on LMD-collected cells. Finally, STR analysis was performed on LMD-separated sperm cells from mixtures of semen and female buccal epithelial cells. The results indicated (a) hematoxylin/eosin staining performed best in its ability to differentiate sperm and epithelial cells while exhibiting the least negative effect on further downstream analysis; (b) both QIAamp and Lyse-N-Go methods were useful for recovery of DNA from LMD-collected sperm cells; and (c) LMD separation provided clear STR profiles of the male donor with the absence of any additional alleles from the female donor. This report describes an efficient, low-manipulation LMD method for the efficient separation of spermatozoa from two-donor sperm/epithelial cell mixtures.

Dental CT imaging as a screening tool for dental profiling: advantages and limitations.

Abstract: The use of dental processing software for computed tomography (CT) data (Dentascan) is described on postmortem (pm) CT data for the purpose of pm identification. The software allows reconstructing reformatted images comparable to conventional panoramic dental radiographs by defining a curved reconstruction line along the teeth on oblique images. Three corpses that have been scanned within the virtopsy project were used to test the software for the purpose of dental identification. In every case, dental panoramic images could be reconstructed and compared to antemortem radiographs. The images showed the basic component of teeth (enamel, dentin, and pulp), the anatomic structure of the alveolar bone, missing or unerupted teeth as well as restorations of the teeth that could be used for identification. When streak artifacts due to metal-containing dental work reduced image quality, it was still necessary to perform pm conventional radiographs for comparison of the detailed shape of the restoration. Dental identification or a dental profiling seems to become possible in a noninvasive manner using the Dentascan software.

Analysis of artificial radiocarbon in different skeletal and dental tissue types to evaluate date of death.

Abstract: Radiocarbon dating, with special reference to the modern bomb-curve, can provide useful information to elucidate the date of death of skeletonized human remains. Interpretation can be enhanced with analysis of different types of tissues within a single skeleton because of the known variability of formation times and remodeling rates. Analysis of radiocarbon content of teeth, especially the enamel in tooth crowns, provides information about the date of formation in the childhood years and in consideration of the known timing of tooth formation can be used to estimate the birth date after 1950 ad. Radiocarbon analysis of modern cortical and trabecular bone samples from the same skeleton may allow proper placement on the pre-1963 or post-1963 sides of the bomb-curve as most trabecular bone generally undergoes more rapid remodeling than does most cortical bone. Pre-1963 bone formation would produce higher radiocarbon values for most trabecular bone than for most cortical bone. This relationship is reversed for formation after 1963. Radiocarbon analysis was conducted in this study on dental, cortical, and trabecular bone samples from two adult individuals of known birth (1925 and 1926) and death dates (1995 and 1959). As expected, the dental results correspond to prebomb-curve values reflecting conditions during the childhoods of the individuals. The radiocarbon content of most bone samples reflected the higher modern bomb-curve values. Within the bone sample analyses, the values of the trabecular bone were higher than those of cortical bone and supported the known placement on the pre-1963 side of the bomb-curve.

Genetic Variation in Hemp and Marijuana (Cannabis sativa L.) According to Amplified Fragment Length Polymorphisms

Abstract: Cannabis sativa L (Cannabaceae) is one of the earliest known cultivated plants and is important in the global economy today as a licit and an illicit crop Molecular markers distinguishing licit and illicit cultivars have forensic utility, but no direct comparison of hemp and marijuana amplified fragment length polymorphism (AFLP) has been made to date Genetic variation was surveyed in three populations of fiber hemp and a potent cultivar of marijuana using AFLP markers Ten primer pairs yielded 1206 bands, of which 88% were polymorphic Eighteen bands represented fixed differences between all fiber populations and the drug cultivar These markers have practical utility for (1) establishing conspiracy in the cultivation and distribution of marijuana, (2) identifying geographic sources of seized drugs, and (3) discriminating illegal, potent marijuana cultivars from hemp where the cultivation of industrial hemp is permitted

Metric assessment of the "mastoid triangle" for sex determination: a validation study.

Abstract: Recently, a metric approach to skeletal sex determination was published by Paiva and Segre which is based on the summation of two triangular areas defined by three distinct craniometric landmarks: Porion, Mastoidale, and Asterion. According to the authors, values for the total triangle > or =1447.40 mm(2) are characteristic for male crania, while values < or =1260.36 mm(2) are indicative of female skulls (95% confidence). In order to evaluate the method's validity, two sex- and age-documented samples of different provenience were analyzed (N=197). The results show that while the indicated measurements display significant sex differences, the technique is of little practical meaning where a single individual must be independently classified. It is hypothesized that differences in the expression of sexual dimorphism as well as a population-specific variability of the asterion location undermine the value of the mastoid triangle as a sex determinant.

Computation of likelihood ratios in fingerprint identification for configurations of three minutiæ

Abstract: Recent challenges to fingerprint evidence have brought forward the need for peer-reviewed scientific publications to support the evidential value assessment of fingerprint. This paper proposes some research directions to gather statistical knowledge of the within-source and between-sources variability of configurations of three minutiae on fingermarks and fingerprints. This paper proposes the use of the likelihood ratio (LR) approach to assess the value of fingerprint evidence. The model explores the statistical contribution of configurations of three minutiae using Tippett plots and related measures to assess the quality of the system. Features vectors used for statistical analysis have been obtained following a preprocessing step based on Gabor filtering and image processing to extract minutia position, type, and direction. Spatial relationships have been coded using Delaunay triangulation. The metric, used to assess similarity between two feature vectors is based on an Euclidean distance measure. The within-source variability has been estimated using a sample of 216 fingerprints from four fingers (two donors). Between-sources variability takes advantage of a database of 818 ulnar loops from randomly selected males. The results show that the data-driven approach adopted here is robust. The magnitude of LRs obtained under the prosecution and defense propositions stresses upon the major evidential contribution that small portions of fingermark, containing three minutiae, can provide regardless of its position on the general pattern.

The Effects of Chemical and Heat Maceration Techniques on the Recovery of Nuclear and Mitochondrial DNA from Bone

Abstract: Forensic anthropologists use a number of maceration techniques to facilitate skeletal analysis of personal identity and trauma, but they may unwittingly eliminate valuable DNA evidence in the process. This study evaluated the effect of 10 maceration methods on gross bone structure and the preservation of DNA in ribs of 12 pigs (Sus scrofa). A scoring system was applied to evaluate the ease of maceration and resulting bone quality while DNA purity was quantified by optical densitometry analysis, followed by polymerase chain reaction (PCR) amplification of three mitochondrial and three nuclear loci. The results demonstrated that while mitochondrial DNA could be amplified for all experiments, cleaning treatments using bleach, hydrogen peroxide, ethylenediaminetetraacetic acid/papain, room temperature water and detergent/sodium carbonate followed by degreasing had low DNA concentrations and failed to generate nuclear PCR products. In general, treatments performed at high temperatures (90 degrees C or above) for short durations performed best. This study shows that traditionally "conservative" maceration techniques are not necessarily the best methods to yield DNA from skeletal tissue.

Review of analytical techniques for arson residues.

Abstract: Arson is a serious crime that affects society through cost, property damage, and loss of life. It is important that the methods and technologies applied by fire investigators in detection of evidence and subsequent analyses have a high degree of reliability, sensitivity, and be subject to rigorous quality control and assurance. There have been considerable advances in the field of arson investigation since the 1950s. Classification of ignitable liquids has been updated to include many new categories due to developments in the petroleum industry. Techniques such as steam or vacuum distillation and gas chromatography (GC) with flame ionization detection that may have been considered acceptable--even a benchmark--40 years ago, are nowadays generally disfavored, to the extent that their implementation may almost be considered as ignorance in the field. The advent of readily available mass spectrometric techniques has revolutionized the field of fire debris analysis, increasing the degree of sensitivity and discrimination possible considerably. Multi-dimensional GC--particularly GC x GC--while not yet widely applied, is rapidly gaining recognition as an important technique. This comprehensive review focuses on techniques and practices used in fire investigation, from scene investigation to analysis.

The Association of Alcohol-Induced Blackouts and Grayouts to Blood Alcohol Concentrations

Abstract: The primary aim of this study was to investigate the association between measured blood alcohol concentration (BAC) and the presence and degree of amnesia (no amnesia, grayout, or blackout) in actively drinking subjects. A secondary aim was to determine potential factors other than BAC that contribute to the alcohol-induced memory loss. An interview questionnaire was administered to subjects regarding a recent alcohol associated arrest with a documented BAC greater than 0.08 g/dL for either public intoxication, driving under the influence, or under age drinking was administered. Demographic variables collected included drinking history, family history of alcoholism, presence of previous alcohol-related memory loss during a drinking episode, and drinking behavior during the episode. Memory of the drinking episode was evaluated to determine if either an alcohol-induced grayout (partial anterograde amnesia) or blackout (complete anterograde amnesia) occurred. Differences in (1) mean total number of drinks ingested before arrest, (2) gulping of drinks, and (3) BAC at arrest were found for those having blackouts compared with no amnesia; while differences in drinking more than planned were found between the no amnesia and grayout groups. A strong linear relationship between BAC and predicted probability of memory loss, particularly for blackouts was obvious. This finding clinically concludes that subjects with BAC of 310 g/dL or greater have a 0.50 or greater probability of having an alcoholic blackout.

The Application of Ultraviolet Irradiation to Exogenous Sources of DNA in Plasticware and Water for the Amplification of Low Copy Number DNA

Abstract: Using high sensitivity forensic STR polymerase chain reaction (PCR) typing procedures, we have found low concentrations of DNA contamination in plasticware and water assumed to be sterile, which is not detected by standard DNA procedures. One technique commonly used to eliminate the presence of DNA is ultraviolet (UV) irradiation; we optimized such a protocol used in the treatment of water, tubes, plates, and tips for low copy number DNA (LCN) amplification. UV light from a Stratalinker((R)) 2400 was administered to 0.2, 1.5 mL tubes, and PCR plates contaminated with up to 500 pg of DNA. They were subsequently quantified with an ALU-based real-time PCR method using the Rotorgene 3000. Overall, there was a decrease in concentration of DNA recovered as the duration of treatment increased. Nonetheless, following 45 min of irradiating a PCR plate with 500 pg of DNA, nearly 6 pg were still detected. However, when the plate was raised within an inch of the UV source, less than 0.2 pg of DNA was detected. Additionally, lining the area around the samples with aluminum foil further reduced the amount of time necessary for irradiation, as only 30 min eliminated the presence DNA in the raised PCR plate. Similar experiments were conducted using tubes filled with a solution of DNA and water in equivalent concentrations for 50, 15, and 1.5 mL tubes with comparative results. It is plausible that the aluminum foil increased the amount of reflection in the area thereby enhancing penetration of UV rays through the walls of the plasticware. This protocol was tested for the possibility of inhibitors produced from irradiation of plastic tubes. As our protocols require less irradiation time than previous studies, PCR sensitivity was not affected. Moreover, the lifespan of the UV lamps was extended. Our findings demonstrate that this method is useful as an additional precautionary measure to prevent amplification of extraneous DNA from plasticware and water without compromising the sensitivity of LCN DNA amplifications.

Simultaneous determination of total human and male DNA using a duplex real-time PCR assay.

Abstract: A single duplex assay to determine both the amount of total human DNA and the amount of male DNA in a forensic sample has been developed. This assay is based on TaqMan technology and uses the multicopy Alu sequence to quantitate total human DNA and the multicopy DYZ5 sequence to quantitate Y chromosomal (male) DNA. The assay accepts a wide concentration range of input DNA (2 muL of 64 ng/microL to 0.5 pg/microL), and also allows detection of PCR failure. The PCR product sizes Alu (127 bp) and DYZ5 (137bp) approximate that of the smaller short tandem repeats (STRs) which should make the assay predictive of STR success with degraded DNA. The assay was optimized for probe/primer concentrations and BSA addition and validated on its reproducibility, on its human specificity, on its nonethnic variability, for artificial mixtures and adjudicated casework, for the effect of inhibitors and for state of DNA degradation. This assay should prove very usual in forensic analyses because knowing the relative amounts of male versus female DNA can allow the examiner to decide which samples may yield the most probative value in a case or direct the samples to methods that would yield the greatest information.

Changing patterns of drug and alcohol use in fatally injured drivers in Washington State.

Abstract: We have previously reported on patterns of drug and alcohol use in fatally injured drivers in Washington State. Here we revisit that population to examine how drug use patterns have changed in the intervening 9 years. Blood and serum specimens from drivers who died within 4 h of a traffic accident between February 1, 2001, and January 31, 2002, were analyzed for illicit and therapeutic drugs and alcohol. Drugs when present were quantitated. Samples suitable for testing were obtained from 370 fatally injured drivers. Alcohol was detected above 0.01 g/100 mL in 41% of cases. The mean alcohol concentration for those cases was 0.17 g/100 mL (range 0.02-0.39 g/100 mL). Central nervous system (CNS) active drugs were detected in 144 (39%) cases. CNS depressants including carisoprodol, diazepam, hydrocodone, diphenhydramine, amitriptyline, and others were detected in 52 cases (14.1%), cannabinoids were detected in 47 cases (12.7%), CNS stimulants (cocaine and amphetamines) were detected in 36 cases (9.7%), and narcotic analgesics (excluding morphine which is often administered iatrogenically in trauma cases) were detected in 12 cases (3.2%). For those cases which tested positive for alcohol c. 40% had other drugs present which have the potential to cause or contribute to the driver's impairment. Our report also considers the blood drug concentrations in the context of their interpretability with respect to driving impairment. The data reveal that over the past decade, while alcohol use has declined, some drug use, notably methamphetamine, has increased significantly (from 1.89% to 4.86% of fatally injured drivers) between 1992 and 2002. Combined drug and alcohol use is a very significant pattern in this population and is probably overlooked in DUI enforcement programs.

Lessons learned from large-scale comparative dental analysis following the South Asian tsunami of 2004.

Abstract: The aim of this study was to examine the quality of the ante-(AM) and postmortem (PM) dental data that were submitted for entry into the PLASS data system in Phuket, Thailand, following the Boxing Day (December 26) Tsunami, 2004. The investigators were two forensic odontologists who were part of the New Zealand Disaster Victim Identification team that worked at Wat Yang Yao morgue and at the Information Management Center in Phuket. Our findings underline the usefulness of dental data in human identification, but point to a number of significant sources of error. Of the 78 PM records received, only 68% of radiographs and 49% of photos confirmed the accompanying dental charting. This underlines the value, particularly of photographs of the dental arches, in quality control. It also points to a large error component, which may have been due to inexperience of the operators, fatigue, poor conditions in the temporary morgue, or the problem of tooth-colored fillings. Of the 106 AM records received, 62% were of unacceptable quality and 64% were either not accompanied by radiographs or had poor quality radiographs. These results indicate that AM data collection ideally needs to be collated and checked by a forensically trained dentist(s) in the country of origin.

Detection of visible and latent fingerprints using micro-X-ray fluorescence elemental imaging.

Abstract: Using micro-X-ray fluorescence (MXRF), a novel means of detecting fingerprints was examined in which the prints were imaged based on their elemental composition. MXRF is a nondestructive technique. Although this method requires a priori knowledge about the approximate location of a print, it offers a new and complementary means for detecting fingerprints that are also left pristine for further analysis (including potential DNA extraction) or archiving purposes. Sebaceous fingerprints and those made after perspiring were detected based on elements such as potassium and chlorine present in the print residue. Unique prints were also detected including those containing lotion, saliva, banana, or sunscreen. This proof-of-concept study demonstrates the potential for visualizing fingerprints by MXRF on surfaces that can be problematic using current methods.

Metacarpal sexual determination in a Spanish population.

Abstract: Anthropologists and forensic pathologist determine the sex of skeletons by analyzing quantitative and qualitative characters in the bone remains. Generally, the skull and os coxae are the elements most used, but they are not always preserved. In such cases, the investigator needs to have available other techniques based on different remains. The aim of the present work is to develop and describe discriminating functions for sex determination in a recent Spanish population using metacarpal morphology. A sample of bones corresponding to a contemporary Spanish population deposited at the Complutense University of Madrid (UCM) was analyzed. This sample comprised 697 metacarpals, corresponding to 79 adult individuals (37 men and 42 women). These allowed us to obtain 120 unifactorial discriminant functions. We selected the 10 equations, one for each metacarpal from both hands, that provided the best sexual discrimination. The correct sex classification rank progressed from 81%, for right (R) metacarpals IV and V, to 91%, for left (L) metacarpal II. The results suggest that metacarpals are structures that can be used for sex determination in paleoanthropological and forensic identifications.