Showing papers in "Journal of Forensic Sciences in 2002"

Individuality of handwriting.

Abstract: Motivated by several rulings in United States courts concerning expert testimony in general, and handwriting testimony in particular, we undertook a study to objectively validate the hypothesis that handwriting is individual. Handwriting samples of 1500 individuals, representative of the U.S. population with respect to gender, age, ethnic groups, etc., were obtained. Analyzing differences in handwriting was done by using computer algorithms for extracting features from scanned images of handwriting. Attributes characteristic of the handwriting were obtained, e.g., line separation, slant, character shapes, etc. These attributes, which are a subset of attributes used by forensic document examiners (FDEs), were used to quantitatively establish individuality by using machine learning approaches. Using global attributes of handwriting and very few characters in the writing, the ability to determine the writer with a high degree of confidence was established. The work is a step towards providing scientific support for admitting handwriting evidence in court. The mathematical approach and the resulting software also have the promise of aiding the FDE.

Validation of a 16-locus fluorescent multiplex system.

Abstract: STR multiplexes have been indispensable for the efficient genotyping of forensic samples. The PowerPlex 16 System contains the coreCODIS loci, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, D21S11, CSF1PO, FGA, THOI, TPOX, vWA, the sex determinant locus, amelogenin, and two pentanucleotide STR loci, Penta D and Penta E. This multiplex satisfies the locus requirements for most national databases and is the most efficient currently available system due to its single PCR amplification. To provide the groundwork for judicial acceptance, including the publication of primer sequences, and to evaluate laboratory-to-laboratory variation, a developmental validation for casework on this commercially available system was performed in 24 laboratories and produced the following conclusions. Amplification was reliable on a variety of thermal cyclers and product could be analyzed on either an ABI PRISM 310 Genetic Analyzer or an ABI PRISM 377 DNA Sequencer. Genotyping using single source samples was consistent between 0.25 and 2 ng of input DNA template with a few laboratories obtaining complete genotypes at 0.0625 ng. However, heterozygote allele imbalance (<60% peak height balance) caused by stochastic effects was observed at a rate of 13% with 0.125 ng DNA and 22% at 0.0625 ng DNA. Mixture analyses were done using a total of 1 ng of DNA template. Most alleles were detected in mixtures of 4 to 1 and some minor alleles were detected in mixtures of 19 to 1. Optimum amplification cycle number was dependent on the sensitivity of the detection instrument used and could also be adjusted to accommodate larger amounts of DNA on solid supports such as FTA paper. Reaction conditions including volume, annealing temperature, and concentrations of primer, AmpliTaq Gold, and magnesium were shown to be optimal yet robust enough to withstand moderate variations without affecting genotype analysis. Environmental, matrix and standard source analyses revealed an ability to obtain complete genotypes in all sample types except those exposed to 80 degrees C for 12-48 days. Finally, comparison of genotype results from the PowerPlex 16 System with other commercially available systems on non-probative reference and forensic samples showed consistent results.

Trace DNA: a review, discussion of theory, and application of the transfer of trace quantities of DNA through skin contact.

Abstract: Advances in STR PCR DNA profiling technology allow for the analysis of minute quantities of DNA. It is frequently possible to obtain successful DNA results from cellular material transferred from the skin of an individual who has simply touched an object. Handling objects, such as weapons or other items associated with a crime, touching surfaces, or wearing clothing, may represent sufficient contact to transfer small numbers of DNA bearing cells, or trace DNA, which can be successfully analyzed. With this minimal amount of contact required to yield a suspect profile comes tremendous crime solving potential, and a number of considerations for prudent application, and the maximization of evidentiary value. Evidentiary materials not previously considered must be recognized and preserved, and the resulting DNA type profiles interpreted in their proper forensic context.

Decomposition chemistry of human remains: a new methodology for determining the postmortem interval.

Abstract: This study was conducted to characterize the chemistry associated with the decomposition of human remains with the objective of identifying time-dependent biomarkers of decomposition. The purpose of this work was to develop an accurate and precise method for measuring the postmortem interval (PMI) of human remains. Eighteen subjects were placed within a decay research facility throughout a four-year time period and allowed to decompose naturally. Field autopsies were performed and tissue samples were regularly collected until the tissues decomposed to the point where they were no longer recognizable (encompassing a cumulative degree hour (CDH) range of approximately 1000 (approximately 3 weeks)). Analysis of the biomarkers (amino acids, neurotransmitters, and decompositional by-products) in various organs (liver, kidney, heart, brain, muscle) revealed distinct patterns useful for determining the PMI when based on CDHs. Proper use of the methods described herein allow for PMIs so accurate that the estimate is limited by the ability to obtain correct temperature data at a crime scene rather than sample variability.

The Accuracy and Precision of Third Molar Development as an Indicator of Chronological Age in Hispanics

Abstract: The accuracy and precision of chronological age estimation based on the stages of third molar tooth development was studied in a sample of 679 radiographs from individuals of Hispanic origin. The age range was 14.0 to 25.0 years. Eight raters from the University of Texas Health Science Center at Houston Dental Branch evaluated the radiographs according to Demirjian's schematic definitions of crown and root formation. The objective of this study was to evaluate the chronology of third molar development in Hispanics following the protocol of a previous study. Within the Hispanic population, the rate of male third molar development is greater than that of female third molar development for ten stages of crown-root formation. Also within this Hispanic population sample, the rate of maxillary third molar development is ahead of mandibular third molar development. The mean absolute difference between chronological age and estimated age was +/-3.0 years in females and +/- 2.6 years in males.

Chemical composition of fingerprints for gender determination.

Abstract: This work investigates the chemical nature of fingerprints to ascertain whether differences in chemical composition or the existence of chemical markers can be used to determine personal traits, such as age, gender, and personal habits. This type of information could be useful for reducing the pool of potential suspects in criminal investigations when latent fingerprints are unsuitable for comparison by traditional methods. Fin-gertip residue that has been deposited onto a bead was extracted with a solvent such as chloroform. Samples were analyzed by gas chromatogra-phy/ mass spectrometry (GC/MS). The chemical components identified include fatty acids, long chain fatty acid esters, cholesterol and squalene. The area ratios of ten selected components relative to squalene were calculated for a small preliminary experiment that showed a slight gender dif-ference for three of these components. However, when the experiment was repeated with a larger, statistically designed experiment no significant differences between genders were detected for any of the component ratios. The multivariate Hotelling's T2 test that tested all ten-component ratios simultaneously also showed no gender differences at the 5% significance level.

Variability of the pattern of aging on the human skeleton: evidence from bone indicators and implications on age at death estimation.

Abstract: Age at death assessment of adult skeletons is one of the most difficult problems in forensic and physical anthropology Two fundamental sources of error are described: the complex variability in the process of skeletal aging and methodological bias Taking into account these limits, we developed a new scoring system for the auricular surface of the ilium and the pubic symphysis In order to address a large variability, we examine reference samples from Europe, North America, Africa, and Asia Data were processed using Bayesian prediction in order to classify specimens in age range categories Results show that combining indicators do not perform better than the auricular surface used as a single indicator Morphological changes with aging are variable between Asian, African, and European populations, confirming the necessity to use population-specific standards Bayesian prediction produces reliable classification and is applicable for subjects over 50 years old, a real methodological improvement

The use of SEM/EDS analysis to distinguish dental and osseus tissue from other materials.

Abstract: With increasing frequency, relatively small, fragmentary evidence thought to be osseous or dental tissue of human origin is submitted to the forensic laboratory for DNA analysis with the request for positive identification. Prior to performing DNA analysis, however, it is prudent to first perform a presumptive test or "screen" to determine whether the questioned material may be eliminated from further consideration. When material is shown not to be consistent with bone/teeth, DNA testing is not performed. When such determinations cannot be made from gross morphological features, elemental analysis can be indicative. This presumptive test is made possible by applying scanning electron microscopy/energy dispersive X-ray spectroscopy (SEM/EDS) in conjunction with an X-ray spectral database recently developed by the FBI laboratory. This database includes spectra for many different materials including known examples of bone and tooth from many different contexts and representing the full range of taphonomic conditions. Results of SEM/EDS analysis of evidence can be compared to these standards to determine if they are consistent with bone and/or tooth and, if not, then what the material might represent. Analysis suggests that although the proportions and amounts of calcium and phosphorus are particularly important in differentiating bone and tooth from other materials, other minor differences in spectral profile can also provide significant discrimination. Analysis enables bone and tooth to be successfully distinguished from other materials in most cases. Exceptions appear to be ivory, mineral apatite, and perhaps some types of corals.

Application of Lamendin's Adult Dental Aging Technique to a Diverse Skeletal Sample

Abstract: Lamendin et al. (1) proposed a technique to estimate age at death for adults by analyzing single-rooted teeth. They expressed age as a function of two factors: translucency of the tooth root and periodontosis (gingival regression). In their study, they analyzed 306 singled rooted teeth that were extracted at autopsy from 208 individuals of known age at death, all of whom were considered as having a French ancestry. Their sample consisted of 135 males, 73 females, 198 whites, and 10 blacks. The sample ranged in age from 22 to 90 years of age. By using a simple formulae (A = 0.18 x P + 0.42 x T + 25.53, where A = Age in years, P = Periodontosis height x 100/root height, and T = Transparency height x 100/root height), Lamendin et al. were able to estimate age at death with a mean error of +/- 10 years on their working sample and +/- 8.4 years on a forensic control sample. Lamendin found this technique to work well with a French population, but did not test it outside of that sample area. This study tests the accuracy of this adult aging technique on a more diverse skeletal population, the Terry Collection housed at the Smithsonian's National Museum of Natural History. Our sample consists of 400 teeth from 94 black females, 72 white females, 98 black males, and 95 white males, ranging from 25 to 99 years. Lamendin's technique was applied to this sample to test its applicability to a population not of French origin. Providing results from a diverse skeletal population will aid in establishing the validity of this method to be used in forensic cases, its ideal purpose. Our results suggest that Lamendin's method estimates age fairly accurately outside of the French sample yielding a mean error of 8.2 years, standard deviation 6.9 years, and standard error of the mean 0.34 years. In addition, when ancestry and sex are accounted for, the mean errors are reduced for each group (black females, white females, black males, and white males). Lamendin et al. reported an inter-observer error of 9+/-1.8 and 10+/-2 sears from two independent observers. Forty teeth were randomly remeasured from the Terry Collection in order to assess an intra-observer error. From this retest, an intra-observer error of 6.5 years was detected.

Forensic handwriting examiners' expertise for signature comparison.

Abstract: This paper reports on the performance of forensic document examiners (FDEs) in a signature comparison task that was designed to address the issue of expertise. The opinions of FDEs regarding 150 genuine and simulated questioned signatures were compared with a control group of non-examiners' opinions. On the question of expertise, results showed that FDEs were statistically better than the control group at accurately determining the genuineness or non-genuineness of questioned signatures. The FDE group made errors (by calling a genuine signature simulated or by calling a simulated signature genuine) in 3.4% of their opinions while 19.3% of the control group's opinions were erroneous. The FDE group gave significantly more inconclusive opinions than the control group. Analysis of FDEs' responses showed that more correct opinions were expressed regarding simulated signatures and more inconclusive opinions were made on genuine signatures. Further, when the complexity of a signature was taken into account, FDEs made more correct opinions on high complexity signatures than on signatures of lower complexity. There was a wide range of skill amongst FDEs and no significant relationship was found between the number of years FDEs had been practicing and their correct, inconclusive and error rates.

A test of the phenice method for the estimation of sex.

Abstract: The Phenice method for the estimation of sex of an individual from the pelvis was tested on 198 individuals of known sex from the Terry collection by an individual with training in the technique, but no additional training in forensic anthropology. Sex was correctly estimated in 88.4% of all individuals and more accurately estimated in females than in males. In a subsequent test of the same individuals by the same investigator utilizing additional nonmetric pelvic indicators, overall accuracy increased to 96.5%, with slightly greater accuracy for males. The study suggests that experience likely contributes to the accuracy of the Phenice method and that for inexperienced investigators, consultation of all pelvic indicators offers an advantage over using just the three variables in the Phenice method.

Interpreting small quantities of DNA: the hierarchy of propositions and the use of Bayesian networks.

Abstract: The dramatic increase in the sensitivity of DNA profiling systems that has occurred over recent years has led to the need to address a wider range of interpretational problems in forensic science. The issues surrounding questions of the kind "whose DNA is this?" have been the subject of considerable controversy but now it is clear that the emphasis is shifting to questions of the kind "how did this DNA get here?" Such issues are discussed in this paper and new insights are provided by two particular recent developments. First, the notion of the "hierarchy of propositions" that has arisen from a project called Case Assessment and Interpretation (CAI) that has been running in the British Forensic Science Service (FSS). Second, a technique for drawing inferences in the face of many interacting considerations, known as "Bayesian networks"--or "Bayes' nets" for short--that has been the subject of an earlier paper in this journal (1). The discussion is carried out by means of case studies, based on actual cases. It is clear that, whereas the inference in relation to the source of the DNA in a crime sample might be overwhelmingly strong, the inference in relation to the propositions that a jury must consider relating to the identity of the actual offender may be much more tentative.

Skeletal identification using the frontal sinus region: a retrospective study of 39 cases.

Abstract: The importance of identification using the frontal sinus has been previously demonstrated in case reports. In this study, 39 cases of identification using frontal sinus comparison from the Ontario Chief Coroner's Office were reviewed and differences between antemortem and postmortem radiographs examined. All cases involved decedents older than twenty years. Three cases were rejected due to poor antemortem and postmortem film quality. One subject had no frontal sinus. Thirty-five cases provided conclusive postmortem to antemortem pattern matches. Sixteen cases also yielded metric (quantitative) matches. Duration between antemortem and postmortem radiographic examinations, age, gender, and cause of death did not affect the ability to obtain a match. This is the largest study undertaken on actual cases and demonstrates the validity of frontal sinus pattern matching for forensic identification.

Evaluation of mRNA markers for the identification of menstrual blood.

Abstract: The detection of epithelial cells in dried bloodstains by reverse transcriptase-polymerase chain reaction is based on cell- and tissue-specific gene expression. In this paper mRNA markers suitable for the identification of menstrual blood were evaluated. RNA isolated from autopsy tissue samples including endometrium, vaginal mucosa, and blood were screened for tissue-specific expression patterns using RT-PCR with primers for hormone receptors, intermediate filaments, matrix metalloproteinases, heat shock proteins, cytokines, and growth factors. Matrix metalloproteinase (MMP) mRNA could be detected in endometrium but not in blood and other epithelia. This was confirmed in further studies with artificial menstrual bloodstains, indicating that the detection of MMP expression in bloodstains may serve as a forensic marker for menstrual blood.

The use of oral fluid and sweat wipes for the detection of drugs of abuse in drivers.

Abstract: Blood, urine, oral fluid (by spitting or with a Salivette), and sweat samples (by wiping the forehead with a fleece moistened with isopropanol) were obtained from 180 drivers who failed the field sobriety tests at police roadblocks. With quantitative GC-MS, the positive predictive value of oral fluid was 98, 92, and 90% for amphetamines, cocaine, and cannabis respectively. The prevalence of opiate positives was low. The proposed SAMHSA cut-off values for oral fluid testing at the workplace, proved their usefulness in this study. The positive predictive value of sweat wipe analysis with GC-MS was over 90% for cocaine and amphetamines and 80% for cannabis. The accuracy of Drugwipe was assessed by comparing the electronic read-out values obtained on-site after wiping the tongue and the forehead, with the corresponding GC-MS results in plasma, oral fluid, and sweat. The accuracy was always less than 90% except for the amphetamine-group in sweat.

Population-specific histological age-estimating method: a model for known African-American and European-American skeletal remains.

Abstract: Previously developed histological age-estimating methods have been based on samples lacking interpopulation variability. A comparison of age-associated rib histomorphometrics between an European-American sample and an African-American sample indicates that ethnicity can have a significant effect on osteon population density (OPD), osteon cross-sectional area (On.Ar), and relative cortical area (Ct.Ar/Tt.Ar). Based upon these findings, new histological age-predicting formulae are presented that are recommended when estimating age for African-American or European-American skeletal remains. A general formula that is applicable to remains of unknown ethnicity is also provided.

Time of submergence using aquatic invertebrate succession and decompositional changes.

Abstract: Pig carcasses were placed in pond and stream habitats in the Malcolm Knapp Research Forest in Maple Ridge, B.C. for approximately one year, to examine the development, species, and sequence of invertebrates associated with the carrion. An invertebrate successional database was created for pond and stream habitats for potential use in estimating time of submergence in water related death investigations. Analysis has shown that a predictable succession of invertebrates colonize the carrion. However, whether or not this succession is carrion dependent or seasonal is unknown. There is a difference in the species composition between pond and stream habitats. Habitats influence invertebrate fauna, therefore, species colonizing carrion are habitat-specific. In both habitats, no one organism can determine time of submergence alone. Decompositional descriptions from this research were compared with 15 freshwater related death investigations. Similarities were seen in the earlier decompositional characteristics including bloat, discoloration, and nail shedding; however, the human descriptions were so vague that they had little value in determining time of submergence and hence time of death.

Forensic comparison of soils by bacterial community DNA profiling.

Abstract: This preliminary investigation has shown that a soil microbial community DNA profile can be obtained from the small sample of soil recovered from the sole of a shoe, and from soil stains on clothing. We have also shown that these profiles are representative of the site of collection and therefore could potentially be used as associative evidence to prove a link between suspects and crime scenes. Soil community profiles were obtained using the T-RFLP fingerprinting method that uses fluorescent primer technology and semi-automated analysis techniques similar to those used in human DNA profiling in forensic laboratories.

Charred Body: Virtual Autopsy with Multi-slice Computed Tomography and Magnetic Resonance Imaging

Abstract: The correct examination of a charred body is a forensic challenge. Examination, interpretation, and conclusion in respect to identification, vital reactions, toxicological analysis, and determining cause and manner of death are all more difficult than without burns. To evaluate what can be seen in the case of a charred body, we made an examination with the new radiological modalities of cross-section techniques, via multi-slice Computed Tomography (MSCT) and Magnetic Resonance Imaging (MRI), prior to performing the classical forensic autopsy. In a charred body case of a single motor vehicle/fixed object collision with a post crash fire, the radiological methods of MSCT and MRI made it possible to document the injuries caused by burn as well as the forensic relevant vital reactions (air embolism and blood aspiration). In conclusion, we think postmortem imaging is a good forensic visualization tool with a great potential for the forensic documentation and examination of charred bodies.

Interobserver variation of selected postcranial skeletal measurements.

Abstract: Osteometric data are of great importance for analytical purposes in the field of forensic anthropology, and it is critical that interobserver concordance is high in order for the results of these analyses to be reliable. Significant interobserver variation of skeletal measurements is cause for concern since it may result in conflicting conclusions. The range of interobserver variation of selected postcranial measurements is addressed. Thirteen standard measurements familiar to most forensic anthropologists were examined, as well as nine nonstandard measurements that were unfamiliar to most participants in the study. Sixty-eight individuals participated in the study, the majority of whom considered themselves to be forensic anthropologists with various levels of experience. In general, interobserver variation of the tested standard postcranial measurements was found to be minimal, with the exception of pubis length and subtrochanteric femur measurements. The difficulties that can lead to measurement error are discussed and possible solutions are recommended.

In vivo facial tissue depth measurements for white British children.

Abstract: This paper reports the results of a study of facial tissue depth measurement in White British children of both sexes, aged between 11 and 18 years. The purpose of this research was to increase the information available upon tissue depth data for children, primarily for use in forensic facial reconstruction. Facial tissue depths were measured at 21 anatomical points using ultrasonic echo-location. The mid-philtral, upper lip border and lower lip border points showed consistently larger tissue depths in the males than the females, and the zygomatic attachment showed consistently larger tissue depths in the females than the males. The males showed a general increase in tissue depth with an increase in age at all the mid-line facial points and the cheek points. The females showed increased tissue depth with age at all the points except the infra-orbital, lateral orbital, mid-zygomatic arch and mid-mandibular points. A table of mean tissue depths was developed for males and females divided into two-yearly age groups.

New formulae for estimating stature in the Balkans.

Abstract: Recent studies of secular change and allometry have observed differential limb proportions between the sexes, among and within populations. These studies suggest that stature prediction formulae developed from American Whites may be inappropriate for European populations. The purpose of this investigation is to present more appropriate stature prediction equations for use in the Balkans to aid present-day identifications of the victims of genocide. The reference sample totals 545 white males obtained from World War II data. The Eastern European sample totals 177 males and includes both Bosnian and Croatian victims of the recent war. Mean stature for Eastern Europeans was obtained from the literature. Results show that formulae based on Trotter and Gleser systematically underestimate stature in the Balkans. Because Eastern Europeans are taller than American Whites it is appropriate to use this as an "informative prior" that can be applied to future cases. This informative prior can be used in predictive formulae, since it is probably similar to the sample from which the Balkan forensic cases were drawn. Based on Bayes' Theorem new predictive stature formulae are presented for Eastern Europeans.

Correlation of microscopic and mitochondrial DNA hair comparisons.

Abstract: Expert opinions regarding the microscopic comparison of human hairs have been accepted routinely in courts for decades. However, with the advent of mitochondrial DNA (mtDNA) sequencing, an assessment can be made of the association by microscopic hair comparisons in casework between a questioned hair and reference hairs from an individual. While each method can be used separately, the two analytical methods can be complementary and together can provide additional information regarding source association. Human hairs submitted to the FBI Laboratory for analysis between 1996 and 2000 were reviewed. Of 170 hair examinations, there were 80 microscopic associations; of these, only nine were excluded by mtDNA. Importantly, 66 hairs that were considered either unsuitable for microscopic examinations or yielded inconclusive microscopic associations provided mtDNA results. Only six hairs did not provide sufficient mtDNA, and only three yielded inconclusive results. Consistency was observed in exculpatory results with the two procedures. This study demonstrates the utility of microscopic hair examinations and the strength of combining microscopic analysis with mtDNA sequencing.

Brake linings: a source of non-GSR particles containing lead, barium, and antimony.

Abstract: The observation of environmental particles similar in composition to gunshot residue (GSR) are not new to forensic experts and have been described in the scientific literature. In order to better define the origin of these particles, brake linings and their wear products were examined by SEM-EDX. The results obtained demonstrate that some types of brake linings contain lead, barium, and antimony and that they can represent a source of particles showing GSR-like elemental profiles. Most of these particles can be easily discriminated from primer discharge residue because of the high levels of iron or the presence of "prohibited" elements in the spectrum. However, particles with iron at minor or trace levels and lacking "prohibited" elements were also found. It is thus advisable to use caution when describing the composition of similar particles as "unique" to primer discharge residue. The strict application of a rigorous morphological criterion is also recommended.

Classification of narcotics in solid mixtures using principal component analysis and Raman spectroscopy.

Abstract: Eighty-five solid samples consisting of illegal narcotics diluted with several different materials were analyzed by near-infrared (785 nm excitation) Raman spectroscopy. Principal Component Analysis (PCA) was employed to classify the samples according to narcotic type. The best sample discrimination was obtained by using the first derivative of the Raman spectra. Furthermore, restricting the spectral variables for PCA to 2 or 3% of the original spectral data according to the most intense peaks in the Raman spectrum of the pure narcotic resulted in a rapid discrimination method for classifying samples according to narcotic type. This method allows for the easy discrimination between cocaine, heroin, and MDMA mixtures even when the Raman spectra are complex or very similar. This approach of restricting the spectral variables also decreases the computational time by a factor of 30 (compared to the complete spectrum), making the methodology attractive for rapid automatic classification and identification of suspect materials.

Ecstasy (MDMA) deaths in New York City: a case series and review of the literature.

Abstract: MDMA ("ecstasy") has gained renewed popularity as a drug of abuse. To access the epidemiology and causes of death of MDMA-positive fatalities, all deaths investigated by the OCME that tested positive for MDMA (22 deaths) between January 1997 and June 2000 were reviewed. There were three deaths in each 1997 and 1998, eleven in 1999, and five in the first part of 2000. Of these 22 deaths, 13 were due to acute drug intoxications, 7 due to mechanical injury (blunt trauma, gunshot wounds), and 2 due to a combination of natural disease and acute drug intoxication. Evidence of recent opiate and/or cocaine use was found in 7 of the acute intoxication deaths and in none of the traumatic or combination natural/intoxication deaths. The race of all decedents was White between the ages of 17-41 years, and 18 of 22 were men.

An improved method for post-PCR purification for mtDNA sequence analysis

Abstract: Mitochondrial DNA (mtDNA) analysis of forensic samples typically is performed when the quantity and quality of DNA are insufficient for nuclear DNA analysis or when maternal relatives may be the only reference source. Many of the steps required in the analytical process are both lengthy and labor intensive. Therefore, improvements in the process that reduce labor without compromising the quality of the data are desirable. The current procedure requires purification of the amplicons by centrifugal filtration after PCR and prior to cycle sequencing. Because this method requires several manipulations to perform, alternate cleanup procedures were investigated. These include the use of 1) Qiagen QlAquick PCR Purification columns, 2) Concert Rapid PCR Purification columns, and 3) ExoSAP-IT reagent. When the yield of purified amplicons, quality of the sequence profile, and ease of assay were evaluated, the use of ExoSAP-IT reagent for post-amplification purification was chosen to replace the filtration method.

Twgdam Validation of Ampf_str•: PCR Amplification Kits for Forensic DNA Casework

Abstract: Laboratory procedures used in short tandem repeat (STR) analysis were subjected to various scenarios that assessed reliability and identified potential limitations. These validation studies were designed as recommended by the Technical Working Group on DNA Analysis Methods (TWGDAM) and the DNA Advisory Board (DAB) (17,18). Various DNA samples were amplified by the polymerase chain reaction (PCR) using AmpFlSTR PCR Amplification Kits (i.e., AmpFlSTR Green I, Profiler, Profiler Plus, and COfiler kits), detected with ABI Prism instrumentation, and analyzed using GeneScan and Genotyper software. Data acquired in these studies reinforced an existing body of knowledge and expertise regarding application and interpretation of STR typing in the forensic science community. Consistent STR genotypes were detected in various body tissues and fluids. Inter-laboratory comparisons produced concordant genotype results. Quantitative interpretational aids for DNA mixtures were characterized. Ability of the typing systems to type potentially compromised samples reliably was evaluated. Nonprobative case evidentiary DNA was successfully amplified, genotyped, and interpreted. Potential limitations or cautionary factors in the interpretation of minimal fluorescence intensity were demonstrated. Differential amplification between loci was observed when PCR was inhibited; preferential amplification typically was not. Single AmpFlSTR locus amplification did not offer consistent benefit over AmpFlSTR multiplexing, even in cases of DNA degradation or PCR inhibition. During rigorous evaluation, AmpFlSTR PCR Amplification Kits reproducibly yielded sensitive and locus-specific results, as required in routine forensic analyses.

Effect of temperature on development of Liopygia (= Sarcophaga) argyrostoma (Robineau-Desvoidy) (Diptera: Sarcophagidae) and its forensic implications.

Abstract: The temperature-dependent development of the forensically important flesh fly Liopygia (Thomsonea) argyrostoma (Robineau-Desvoidy) (=Sarcophaga argyrostoma) was studied at six constant temperature regimes in the laboratory. Total developmental time of L. argyrostoma from larviposition to adult emergence was 54.9+/-1.45, 31.3+/-1.1, 22.2+/-0.67, 16.3+/-0.54, and 14.9+/-0.4 days (+/-S.D.) when reared at 15, 20, 25, 30, and 35 degrees C, respectively. At 8 degrees C, larval development was not completed. From linear regression of development rates of five of the six studied constant temperature regimes, it was determined that the minimum development threshold (tL) for total immature development is 7.4 degrees C, and the overall thermal constant (K) for L. argyrostoma is 396.4+/-19.18 (mean +/- S.D.) day-degrees (DD) above the threshold.

Detection of petrol (gasoline) in fire debris by Gas chromatography/mass spectrometry/mass spectrometry (GC/MS/MS)

Abstract: Gas chromatography/mass spectrometry (GC/MS) is a well-known instrumental technique used for the analysis of fire debris for accelerant detection. However, matrix problems, such as pyrolysis product interference, are still encountered. These interferences often lead to inconclusive interpretation of the chromatographic results obtained. This paper describes a method of analyzing arson accelerants using gas chromatography coupled with ion trap mass spectrometry/mass spectrometry (GC/MS/MS). Petrol was investigated as an accelerant. Analysis incorporated an ion preparation method (IPM) that manipulates the ion population in the ion trap following ionization but prior to ion analysis. Parent ions of selected mass-to-charge (m/z) values, characteristic for petrol, are stored within the ion trap. All other unwanted ions are physically removed from the trap by sophisticated electrostatic waveforms. The parent ions fill the trap and are then fragmented by means of collision-induced dissociation (CID). The resultant daughter ion spectra are obtained, which can be used for identification purposes. In this manner, ions of pyrolysis origin are removed.