The use of crime scene detection dogs to locate semen stains on different types of fabric.

In sexual assault cases, the detection and identification of semen is extremely important as this type of evidence can be used as a source for investigative leads and contributes to case evidence. However, the detection of semen stains is often difficult, even indoors, because of different (environmental) factors, such as substrate type, coloured items and large search areas. In 2015, a project was initiated by the Dutch police to evaluate the feasibility of the use of detection dogs to locate semen stains in forensic practise. Since promising results were obtained, here, a double-blind study was designed to investigate how these detection dogs can optimally be implemented in the current work flow of crime scene investigators and to compare the dog's sensitivity and specificity with current detection methods. The performance of the detection dogs was compared to three commonly used detection methods for semen, (i) forensic light sources (FLS), (ii) the RSID semen field kit and (iii) the enzymatic Acid Phosphatase (AP)-test on semen deposited at different types of fabrics. A 100% sensitivity and specificity for the detection of semen stains using the detection dogs was obtained, compared to an overall sensitivity and specificity of 76.3% and 100% for FLS, 81.6% and 100% for RSID-test, and 92.1% and 100% for AP-test, respectively. Especially, on fabrics with a pattern or interfering fluorescent properties, detection dogs demonstrated to be of additional value to locate the semen stains. We recommend to use the following order of testing, FLS, detection dog, AP-test and RSID test in a forensic workflow.

White light reflectance spectroscopy biosensing system for fast quantitative prostate specific antigen determination in forensic samples.

A label-free biosensor based on white light reflectance spectroscopy for the determination of PSA as semen indicator in forensic samples is presented. The sensor is based on a two-step immunoassay which employs the same polyclonal anti-PSA antibody as capture and detection antibody followed by reaction with streptavidin as a signal enhancement step. The whole assay time was set to 10min; 5min reaction of immobilized antibody with the PSA calibrators or the samples, 3min reaction with the biotinylated anti-PSA antibody and 2min reaction with streptavidin. Following this protocol, a detection limit of 0.5ng/mL was achieved and the assay's linear response range extended up to 500ng/mL. Thus, taking into account the quantification limit of 1.0ng/mL and the average PSA concentration in semen (0.2-5.5mg/mL), semen quantities of a few nanoliters could be detected. The accuracy of the sensor developed was demonstrated through recovery (% recovery ranged from 89.6 to 106) and semen dilution experiments. A linear correlation was found for semen dilutions ranging from 5000 to 360,000. The lack of interference by other bodily fluids was confirmed by analysing stains of blood, urine and saliva prior to and after the addition of semen. Finally, the sensor was evaluated by analysing 51 forensic casework samples which were also analysed with a semi-quantitative membrane strip test (Seratec® PSA), through microscopic detection of spermatozoa, and male DNA identification through detection of Y chromosome. The results obtained with the sensor were in excellent agreement with those provided by an immunoradiometric assay kit (PSA-RIACT) and in complete agreement with the findings using the membrane strip assay, spermatozoa and Y chromosome detection. The excellent analytical performance and small size of the instrument make the sensor developed an attractive tool for use in forensic evidence screening for semen detection.

In Situ Identification of Semen Stains on Common Substrates via Raman Spectroscopy,.

The spectroscopic identification of body fluids in situ is a major objective in forensic science. This approach offers the confirmatory, nondestructive, rapid, and on-scene identification of various body fluids. Although Raman spectroscopy has shown tremendous promise toward this goal in prior proof-of-concept experiments, a significant challenge which still remains is substrate interference. Here, an approach for detecting semen stains in situ on various substrates using Raman spectroscopy is explored. Simulated semen evidence was prepared on skin, glass, and various fabrics. Raman data were accumulated from stains without any pretreatment using a common confocal mapping spectrometer using 785 nm laser excitation. The results demonstrate that the spectroscopic interferences encountered by substrates can be reduced and eliminated using a combination of existing subtraction techniques and chemometric models. Heterogeneous substrates proved most challenging, however, automatic subtraction treatment, and location of fluid hotspots was able to elucidate a clear spectroscopic signature of semen in every instance.

Comparison between prostate specific antigen and acid phosphatase for detection of semen in vaginal swabs from raped women.

OBJECTIVE: To compare the effectiveness of the prostate specific antigen (PSA) test and the acid phosphatase (AP) test for semen detection in human vaginal samples. MATERIAL AND METHOD: The source materials were vaginal swabs that were tested at Ramathibodi Hospital between 2008 and 2010 from 2450 cases of raped women. Each swab was tested for semen by three methods: sperm detection by light microscopy, the AP enzymatic reaction, and the presence of PSA by using an immuno-chromatographic rapid kit test. The efficiencies of the AP and PSA tests were compared using the light microscopy result for the presence of sperm as the gold standard. RESULT: The specificities of the AP, the PSA and the combined AP-PSA tests were 96.4%, 92.3% and 91.9%, respectively, and the sensitivities were 65.5%, 80.4% and 84.5%, respectively. The receiver operating characteristic (ROC) area of the AP, PSA and combined AP-PSA tests were 0.8091, 0.8639 and 0.8823, respectively. The ROC area of the PSA test was significantly greater than that of the AP test (p < 0.0001), and the ROC area of the combined AP-PSA test was significantly greater than both the tests individually (p < 0.0001). DISCUSSION: Based on the ROC area, the PSA test was better than the AP test for semen detection in the vaginal swabs, and the combined results (AP + PSA) were better than the individual tests. The specificity of the AP test was higher than the PSA test in this study because a positive detection was made within only 15 s. While the PSA test was more convenient as it was available in a rapid test kit format, our recommendation is PSA detection should be done together with AP test and spermatozoa examination to identify evidence of rape. CONCLUSION: Using these three tests together (AP, PSA, and spermatozoa detection) was recommended as a forensic tool for investigations of vaginal swabs of the rape victims.

Fundamental issues in forensic semen detection / Problemas fundamentales en la detección forense de semen

Sperm detection can be an important factor in confirming sexual assault in cases of rape. A large number of cases received in a forensic laboratory involve sexual offenses, making it necessary to examine exhibits for the presence of seminal stains. The objective of this paper is to provide an overview of the most important methods and tests used in the identification of spermatozoa or constituents ofseminal fluid during the investigation ofalleged sexual assault cases in forensic medical practice. Furthermore, this paper focusses on the basic knowledge that is necessary to the graduate students who wish to specialize in forensic sciences.

La detección de esperma puede ser un factor importante a la hora de confirmar un ataque sexual en los casos de violación. Un gran número de los casos recibidos en el laboratorio forense tienen relación con ofensas sexuales, lo cual hace necesario examinar muestras de presencia de manchas seminales. El objetivo de este trabajo es proporcionar una apreciación global de los métodos y pruebas más importantes usados en la identificación de espermatozoos o constituyentes del fluido seminal durante la investigación de supuestos casos de ataque sexual, en la práctica médica forense. Además, este trabajo presta atención al conocimiento básico necesario para los estudiantes graduados que desean especializarse en las ciencias forenses.