Developmental validation of a novel lateral flow strip test for rapid identification of human blood (Rapid Stain Identification™-Blood)

Human blood is the body fluid most commonly encountered at crime scenes, and blood detection may aid investigators in reconstructing what occurred during a crime. In addition, blood detection can help determine which items of evidence should be processed for DNA-STR testing. Unfortunately, many common substances can cause red-brown stains that resemble blood. Furthermore, many current human blood detection methods are presumptive and prone to false positive results. Here, the developmental validation of a new blood identification test, Rapid Stain Identification™-Blood (RSID™-Blood), is described. RSID™-Blood utilizes two anti-glycophorin A (red blood cell membrane specific protein) monoclonal antibodies in a lateral flow strip test format to detect human blood. We present evidence demonstrating that this test is accurate, reproducible, easy to use, and highly specific for human blood. Importantly, RSID™-Blood does not cross-react with ferret, skunk, or primate blood and exhibits no high-dose hook effect. Also, we describe studies on the sensitivity, body fluid specificity, and species specificity of RSID™-Blood. In addition, we show that the test can detect blood from a variety of forensic exhibits prior to processing for DNA-STR analysis. In conclusion, we suggest that RSID™-Blood is effective and useful for the detection of human blood on forensic exhibits, and offers improved blood detection when compared to other currently used methods.     

Photographic Image Comparison of Vegetation

Images and videos are common types of evidence in crime scene investigations and laboratory analysis. Images may be taken by the suspect and/or by crime scene investigators and may serve as crucial elements in forensic laboratory analysis. Forensic photographic image comparison is the process of comparing one or more objects or persons in photographs/images when at least one image is known to be related to a crime. The forensic examiner usually compares the images in order to determine whether or not an association between the exhibits in the images can be made. This paper proposes an extension to the currently prevalent photographic image examination method. The extension introduces comparison of landscape and vegetation over time. It is revealed that similarities between images may still be found between the period of time the suspect photograph was taken and the period the crime scene investigator took the photograph from the same area. In this case report, two suspects to be involved in growing a marijuana field were arrested by the police. The forensic experts were asked to examine images taken by the crime scene investigators and to compare them to the images found in the suspects’ phones. They then tried to determine whether the suspects could be linked to the specific locations. While applying techniques derived from morphological comparison methodologies, the plants at the scene provided significant additional information. A tree trunk, branches, and twigs on a hedge in the photographs revealed specific individual characteristics that led the examiner to reach a conclusive decision.     

Turf wars: authority and responsibility for the investigation of mass graves

Mass graves are complex products of large-scale crimes. Such scenes pose four conceptual challenges to investigators and forensic experts: the individual victim, the crime, the setting, and the statistical. Exhumation and post-mortem examination of mortal remains with associated personal and forensic evidence require integrated management of core forensic personnel including investigators, archaeologists, anthropologists, odontologists and pathologists, among whom there is overlapping expertise. The key to avoiding competition and ill-will among experts is to recognize that all such experts should be enabled to make known how their expertise matches with the temporal and spatial boundaries of victim, crime and setting. In turn, they should be apprised of where they fit into the overall judicial process and their limits within the investigation. Consequently, each expert requires access to the factual background of the case, to the site and its contents throughout the investigation. Each forensic team member has a responsibility to influence the investigation--throughout its course when possible--to make findings within their areas of expertise, and to make these available to the rest of the team so as to contribute most meaningfully to the aims of the investigation, both forensic and humanitarian. The on-site crime scene manager has an overarching role to enable integrated access to the complete scene and its contents by each forensic expert team member. In other words, the forensic scientist is given access and the ability to influence the investigation while control of evidence from the site as to identity and criminal activity are maintained by the crime scene manager. This contribution is directed at both the crime scene manager and each forensic expert; it describes the essential spatial and temporal parameters of an expert's opinion so as to encourage cooperation, and discourage conflict, within the forensic team.     

Evaluation of amylase testing as a tool for saliva screening of crime scene trace swabs

Amylase testing has been used as a presumptive test for crime scene saliva for over three decades, mainly to locate saliva stains on surfaces. We have developed a saliva screening application for crime scene trace swabs, utilising an amylase sensitive paper (Phadebas^® Forensic Press test). Positive results were obtained for all tested dried saliva stains (0.5–32 μL) with high or intermediate amylase activity (840 and 290 kU/L). Results were typically obtained within 5 min, and all samples that produced DNA profiles were positive. However, salivary amylase activities, as well as DNA concentrations, vary significantly between individuals. We show that there is no correlation between amylase activity and amount of DNA in fresh saliva. Even so, a positive amylase result indicates presence of saliva, and thereby presence of DNA. Amylase testing may be useful for screening in investigations where the number of DNA analyses is limited due to cost, e.g., in volume crime.     

The Effects of Substance Use on Offender Crime Scene Behavior

Abstract:  Substance use has an effect on an individual's propensity to commit acquisitive crime with recent studies showing substance users more likely to leave forensic material at a crime scene. An examination of acquisitive crime solved in Northamptonshire, U.K., during 2006 enabled 70 crime scene behavior characteristics to be analyzed for substance and nonsubstance use offenders. Logistical regression analyses have identified statistically significant crime scene behavior predictors that were found to be either present at or absent from the crime scene when the offender was a substance user. Most significant predictors present were indicative of a lack of preparation by the offender, irrational behavior, and a desire to steal high value, easily disposed of, property. Most significant predictors absent from the crime scene were indicative of more planning, preparation, and execution by the offender. Consideration is given to how this crime scene behavior might be used by police investigators to identify offenders.     

A comprehensive study into false positive rates for ‘other’ biological samples using common presumptive testing methods

The identification of biological fluids or materials in forensic samples is a key requirement in forensic science that relies on chemical and biological based tests, most of which exhibit false positivity. When reporting results from such tests, Forensic Scientists use words such as probable, possible, and likely, without always being able to provide robust support for these conclusions. In collating information about false positive rates for a number of these tests, we found limited research into the cross reactions observed from ‘other’ biological samples in commonly encountered case sample stains. By ‘other’ we mean biological fluids or materials that are not the primary target of the presumptive test being used. Here we carry out a specificity study to fill gaps in the literature for a number of the presumptive chemical, biological and immunochromatographic tests used to presumptively screen for blood, semen and saliva. The tests selected for this study are the widely used tests: Luminol, TMB/Combur³ Test® E, Kastle-Meyer (KM), RSID™ - Blood, ABAcard® HemaTrace®, Acid Phosphatase (AP), ABAcard® p30, RSID™ - Semen, Phadebas® ‘Tube’ Test, Phadebas® ‘Press’ Test, and RSID™ - Saliva tests. Specificity for each of these was tested in known samples, from volunteers, of blood, semen, saliva, urine, sweat, vaginal material, faeces and breast milk, and then false positive rates were determined.     

Validation of presumptive tests for non-human blood using Kastle Meyer and Hemastix reagents

Kastle Meyer and Hemastix reagents are presumptive tests commonly used in forensic casework for the detection of blood, and their suitability has been reviewed in numerous publications. However, studies to date have focused on the validation of these tests on human blood alone, and no published work has looked at the sensitivity, specificity and effect on DNA analysis when using these reagents to presumptively test for animal blood. The aim of this study was to validate the two reagents for use with animal blood, and compare their performance in order to choose the best test based on the circumstances in wildlife crime investigation.The sensitivity, specificity, stability and robustness of the methods were assessed by experiments with dilutions of animal blood (from 1:4 to 1:65536) using direct and indirect (rub) tests, potential interfering substances, blood sources from different species and aged blood. The effects of the two reagents on subsequent DNA analysis were also investigated.During the direct tests, Kastle Meyer showed a higher sensitivity, detecting blood down to a dilution of 1:16,384, one order of magnitude lower than Hemastix. However during the rub test, Hemastix showed a higher sensitivity, detecting blood down to a dilution of 1:64 on porous materials while Kastle Meyer was positive only down to a dilution of 1:16. Moreover, when using the same swab for presumptive testing and DNA extraction, Hemastix testing allowed amplification of a sufficient amount of DNA for species identification at its limit of sensitivity on porous materials (1:64) while Kastle Meyer inhibited most amplification of DNA at its less sensitive limit of 1:16 dilution. On the other hand, Hemastix showed a much lower specificity, producing false positive results when exposed to tomato, potato, rust, avian uric acid, bleach and sink rot, while Kastle Meyer only produced a faint positive reaction from potato. Both tests performed equally well detecting fresh blood of different animal species. The stability test gave comparable results among the tests except for aged fish blood stains, where the Kastle Meyer test performed poorly.Owing to its ease of use, higher sensitivity, and lack of interference with downstream DNA analysis, and despite its reduced specificity compared to Kastle Meyer, the Hemastix method is more appropriate for use in wildlife crime investigations. Positive results would always be confirmed with DNA analysis and the low interference of the reagent will allow the use of a single swab for presumptive testing and DNA sampling.     

Forensic identification of a rapist using unusual evidence.

This case report demonstrates a rape case, where no semen, hair, or fingerprints were left by the perpetrator at the crime scene, but rather uncharacteristic biological and physical evidence in the form of a lollipop and a pair of glasses. Three separate forensic laboratories collaborated using conventional forensic methods of PCR DNA typing, photography, and toolmark comparisons to provide investigators with scientific evidence which in turn was instrumental in bringing a violent criminal to justice. The importance of evaluating each item of evidence and realizing its forensic value is stressed in this case report.     

The Forensic Confirmation Bias: A Comparison Between Experts and Novices

A large body of research has described the influence of context information on forensic decision‐making. In this study, we examined the effect of context information on the search for and selection of traces by students (N = 36) and crime scene investigators (N = 58). Participants investigated an ambiguous mock crime scene and received prior information indicating suicide, a violent death or no information. Participants described their impression of the scene and wrote down which traces they wanted to secure. Results showed that context information impacted first impression of the scene and crime scene behavior, namely number of traces secured. Participants in the murder condition secured most traces. Furthermore, the students secured more crime‐related traces. Students were more confident in their first impression. This study does not indicate that experts outperform novices. We therefore argue for proper training on cognitive processes as an integral part of all forensic education.     

Estimation of short-term postmortem interval utilizing core body temperature: a new algorithm

The use of temperature-based short-term postmortem interval (PMI) estimation methods can be useful to homicide investigators at the scene of a questionable death; however, a number of current PMI estimation techniques have high error rates such that they are of limited utility to law enforcement investigators and forensic professionals. These methods fail to control confounding errors present in individual data. An averages-based method of short term postmortem interval estimation was compared to eight other methods, and found to predict postmortem interval considerably more accurately, and for much longer periods of time.     

Unnatural death investigation in India. A plea for the introduction of a medical examiner system of inquest

The inquest upon a dead body is the basis of the scientific crime investigation and administration of criminal justice. The pivotal issue in any inquest is the initial visualization of the body and its surroundings at the scene of incident by a team of investigators and experts. This must be carried out meticulously, since the subsequent course of the case, and its success or failure in the court of justice, essentially depends upon its proper execution. Unfortunately, such an important step as the inquest is generally the most neglected part of any crime investigation process as it exists in India today. It is quite often left entirely to the inspired guesswork and seasoned experience of the lowest ranking police officers and more often than not to a Head Constable of a police station or substation. Nevertheless, while the coroner's system has proved to be utterly unsuitable for Indian conditions, we have failed to find an alternative solution to the growing challenge of scientific crime investigation in India, by involving the forensic pathologist in inquests held upon dead bodies, by including him in the first crime investigation. A strong case is made for adoption of the medical examiner's system of inquest, wherein the forensic pathologist plays the pivotal role and leads the "unnatural death investigation team."     

A Rapid,Confirmatory Test for Body Fluid Identification,

We have developed a technique that allows investigators to confirm the presence of blood, semen, and/or saliva in a crime scene sample. It is a confirmatory test where multiple samples can be processed in less than an hour, and it is potentially portable, permitting samples to be processed at the crime scene. Samples at a scene giving a positive result can be further processed while those failing to do so may be ignored. There is a large and growing backlog of DNA evidence in the USA, slowing down the criminal justice system. This backlog has continued to grow despite an increase in the ability to process evidence faster. This technique uses quantum dot molecular beacons to test for tissue‐specific RNA species, identifying particular body fluids. We have demonstrated the tissue specificity of molecular beacons for blood, semen, and saliva.     

Why teach crime scene management to forensic university students?

Despite enhanced forensic collaborations between law enforcement agencies and universities, crime scene management remains a domain seen more as technical than scientific, largely carried out by sworn police officers qualified as scene of crime officers (SOCOs), confining forensic graduates to specialized expertise fields. Under such circumstances, we must ask ourselves why and how do we provide a generalised dedicated course to academic pupils who are not primarily intended to join such teams, excepting sometimes to assist them on a specialized field (explosion, arson, etc.)?Currently in Quebec, forensic graduates cannot join crime scene units attached to police forces, mainly because of unions, which argue about the need to be an experienced police officer before qualifying for a crime scene course.Based on the operational experience of the author, who created the foundational graduate forensic programme in Quebec, Canada, this paper will explain why such an academic course is still of high importance, its rationale within an academic curriculum, its goal and its implementation. Challenges are still to be considered, but selected feedback from students who understood that the aim of this course is distinct from their formal support disciplines, encourage such an approach.     

Suicidal Shotgun Wound Employing a Shotgun Barrel,a Shotgun Shell,and a BB*

Abstract: A vast majority of firearms‐related suicides involve the conventional use of a properly functioning, intact firearm. Occasionally, forensic investigators encounter a case wherein the firearm suicide victim employs some form of unconventional use of a weapon, or utilizes an unusual weapon type. In this case report, the authors present an unusual case in which a man committed suicide by using a shotgun shell, a shotgun barrel (separate from the rest of the gun), and a BB. Some confusion as to the cause and manner of death was present during the initial scene investigation. Examination at autopsy revealed the cause of death to be a shotgun wound of the chest and prompted further scene investigation. Therefore, this case serves as another example of the importance of cooperation between scene investigators and forensic pathologists when investigating a death. A review of suicidal firearm cases using unconventional means and/or firearms is presented.     

Forensic Comparison of Soil Samples Using Nondestructive Elemental Analysis

Soil can play an important role in forensic cases in linking suspects or objects to a crime scene by comparing samples from the crime scene with samples derived from items. This study uses an adapted ED‐XRF analysis (sieving instead of grinding to prevent destruction of microfossils) to produce elemental composition data of 20 elements. Different data processing techniques and statistical distances were evaluated using data from 50 samples and the log‐LR cost (C_llr). The best performing combination, Canberra distance, relative data, and square root values, is used to construct a discriminative model. Examples of the spatial resolution of the method in crime scenes are shown for three locations, and sampling strategy is discussed. Twelve test cases were analyzed, and results showed that the method is applicable. The study shows how the combination of an analysis technique, a database, and a discriminative model can be used to compare multiple soil samples quickly.     

Examination of lightbulb filaments after a car crash: difficulties in interpreting the results

A drunk car driver was involved in a fatal crash with a scooter at night. Examination of the lightbulbs from the vehicles revealed some mechanical and analytical incompatibilities. The laboratory's initial observations led to complementary police investigations which finally solved the first incompatibility. The different precautions taken by the technical police investigators on the crime scene finally allowed the forensic scientists to establish the functional state of the lightbulbs during the crash.     

Effective use of forensic science in volume crime investigations: Identifying recurring themes in the literature

New scientific, technological and legal developments, particularly the introduction of national databases for DNA and fingerprints, have led to increased use of forensic science in the investigation of crime. There is an assumption, and in some instances specific assertions, that such developments bring improvements either in broad criminal justice terms or more narrowly in terms of economic or practical efficiencies. The underlying presumption is that the new technological opportunities will be understood and effectively implemented. This research investigates whether such increases in activity have also been accompanied by improvements in the effective use of forensic science. A systematic review of thirty-six reports published (predominantly in England and Wales) since the 1980s, which have considered the use of forensic science in the investigation of volume crimes, was carried out. These reports have identified a number of recurrent themes that influenced how effectively forensic science was used in investigations. The themes identified included forensic knowledge and training of investigators, communication and information exchange between specialists and investigators, timeliness of forensic results, interagency relationships and deployment of crime scene examiner resources. The research findings suggest that these factors continue to hinder the effective use of forensic science despite technological advances and this paper considers their potential causes.     

Validation studies of rapid stain identification-blood (RSID-blood) kit in forensic caseworks

When bloodstains are detected at crime scene using presumptive tests (e.g. luminol, phenolphthalein, leuchomalachite green), it is important to establish the real human nature of each stain. This is possible using confirmatory tests. One of these is rapid stain identification-blood (RISD-blood) a lateral flow immuno-chromatographic strip test format which allows the identification of human blood by detection of glycophorin A, a red blood cell membrane antigen, using two anti-human glycophorin A (GPA) monoclonal antibodies.The aim of this study is to assess the sensitivity of RSID-blood test in old, degraded bloodstains and in some bloodstains previously treated with BlueStar Forensic, a presumptive test which is often used in crime scene investigations to detect latent bloodstains. The genetic analysis of all bloodstains of confirmed human nature was subsequently performed using the AmpF1STR Identifiler PCR Amplification Kit (Applied Biosystems), to validate the possibility of obtain a consistent and reliable DNA typing results.     

Spherical Photography and Virtual Tours for Presenting Crime Scenes and Forensic Evidence in New Zealand Courtrooms

The delivery of forensic science evidence in a clear and understandable manner is an important aspect of a forensic scientist's role during expert witness delivery in a courtroom trial. This article describes an Integrated Evidence Platform (IEP) system based on spherical photography which allows the audience to view the crime scene via a virtual tour and view the forensic scientist's evidence and results in context. Equipment and software programmes used in the creation of the IEP include a Nikon DSLR camera, a Seitz Roundshot VR Drive, PTGui Pro, and Tourweaver Professional Edition. The IEP enables a clear visualization of the crime scene, with embedded information such as photographs of items of interest, complex forensic evidence, the results of laboratory analyses, and scientific opinion evidence presented in context. The IEP has resulted in significant improvements to the pretrial disclosure of forensic results, enhanced the delivery of evidence in court, and improved the jury's understanding of the spatial relationship between results.     

A case of suicidal hanging staged as homicide

Suicides staged as homicides are rarely encountered by crime scene investigators. The case of one such staged homicide is presented in which the victim used restraints during a hanging. No other cases of suicidal hangings staged as homicides could be found in the forensic literature. Similar cases should be reported so additional data can be gained from these deaths to help reveal indicators of suicide rather than homicide.