From Deutsche Zeitschrift to International journal of legal medicine-100 years of legal medicine through the lens of journal articles, Part 4: International journal of legal medicine from 1990 to 2022.

This is the fourth and final paper in a series related to the analysis of articles published in this journal during its first 100 years of activity. This article covers the time span from 1990 to 2022. It is important to note that, given the period covered by this analysis, it does not aim to provide a historical overview but rather an examination of the most recent trends in our discipline compared to the past. Between 1990 (Volume 104) and 2022 (Volume 136), 4004 articles were published in the International Journal of Legal Medicine (IJLM) across 33 volumes. This corresponds to 53% of all the articles published since the launch of the journal. When compared to the period from 1970 to 1990, some categories no longer appear to be as relevant (e.g., sexual medicine, 1 article; social medicine, 0 articles; biography, 3 articles; history, 4 articles). Conversely, the most recent period has shown an increasing importance in forensic genetics (1388 articles) and the emergence of new significant topics that merit their own classification, such as age estimation (286 articles), forensic anthropology (189 articles), forensic imaging (150 articles), and forensic entomology (90 articles).

From Deutsche Zeitschrift to International Journal of Legal Medicine-100 years of legal medicine through the lens of journal articles : Part 2: Deutsche Zeitschrift für die gesamte gerichtliche Medizin from 1948 to 1969.

The interruption of the publication of the Deutsche Zeitschrift für die gesamte gerichtliche Medizin due to the war ended with volume 39 for the years 1948/1949. Until volume 66/1969, the journal appeared unchanged under the historical title. The 912 publications contained in the 28 volumes of these two decades cover topics from the main fields of forensic medicine, but also from related and unrelated disciplines. The topic-specific analysis of the publications shows a shift of the research focus in the German institutes since the post-war period. This is most evident in the decline in the number of publications from the fields of scientific and technical criminalistics as well as forensic psychiatry and psychology. An opposite trend with a significant increase in scientific papers was observed in alcohology, forensic genetics and traffic medicine. While the evaluated publications on most topics contain new findings that are still valid today, the use of blood group characteristics for forensic purposes came to an end as a result of the introduction of DNA analysis.

From Deutsche Zeitschrift to International Journal of Legal Medicine-100 years of legal medicine through the lens of journal articles : Part 3: Zeitschrift für Rechtsmedizin-Journal of Legal Medicine from 1970 to 1990.

From volume 67 (1970) onwards, the journal appeared under the new bilingual title Zeitschrift für Rechtsmedizin-Journal of Legal Medicine. The editorial board was expanded and internationalised. From 1970 to 1990, 1416 articles were published in 36 volumes. 1036 articles were in German and 380 in English. The authors of 411 articles came from non-German-speaking countries. Compared to the periods under review in the first two parts of our article series, the proportion of papers on forensic genetics increased significantly between 1970 and 1990, with a small increase in publications on the identification of unknown dead bodies. An opposite trend was observed in the articles on forensic psychiatry and psychology, sexual medicine and social medicine. This development reflects a further sharpening of the discipline's profile.

Whole-genome sequencing of artificial single-nucleotide variants induced by DNA degradation in biological crime scene traces.

The aim of this study was to identify artificial single-nucleotide variants (SNVs) in degraded trace DNA samples. In a preliminary study, blood samples were stored for up to 120 days and whole-genome sequencing was performed using the Snakemake workflow dna-seq-gatk-variant-calling to identify positions that vary between the time point 0 sample and the aged samples. In a follow-up study on blood and saliva samples stored under humid and dry conditions, potential marker candidates for the estimation of the age of a blood stain (= time since deposition) were identified. Both studies show that a general decrease in the mean fragment size of the libraries over time was observed, presumably due to the formation of abasic sites during DNA degradation which are more susceptible to strand breaks by mechanical shearing of DNA. Unsurprisingly, an increase in the number of failed genotype calls (no coverage) was detected over time. Both studies indicated the presence of artificial SNVs with the majority of changes happening at guanine and cytosine positions. This confirms previous studies and can be explained by depurination through hydrolytic attacks which more likely deplete guanine while deamination leads to cytosine to thymine variants. Even complete genotype switches from homozygote 0/0 genotypes to the opposite 1/1 genotypes were observed. While positions with such drastic changes might provide suitable candidate markers for estimating short-term time since deposition (TsD), 11 markers were identified which show a slower gradual change of the relative abundance of the artificial variant in both blood and saliva samples, irrespective of storage conditions.

Comparing preservation substrates under field conditions for efficient DNA recovery in bone.

Often bones are the only biological material left for the identification of human remains. As situations may occur where samples need to be stored for an extended period without access to cooling, appropriate storage of the bone samples is necessary for maintaining the integrity of DNA for profiling. To simulate DNA preservation under field conditions, pig rib bones were used to evaluate the effects of bone cleaning, buffer composition, storage temperature, and time on DNA recovery from bone samples. Bones were stored in three different buffers: TENT, solid sodium chloride, and ethanol-EDTA, at 20 °C and 35 °C for 10, 20, and 30 days. Bones were subsequently dried and ground to powder. DNA was extracted and quantified. Results show that temperature and storage time have no significant influence on DNA yield. DNA recovery from bones stored in solid sodium chloride or ethanol-EDTA was significantly higher compared to bones stored in TENT, and grinding of bones was facilitated by the extent of dehydration in solid sodium chloride and ethanol-EDTA compared to TENT. Overall, solid sodium chloride was found to be superior over ethanol-EDTA; when it comes to transportation, dry material such as salt eliminates the risk of leaking; it is non-toxic and in contrast to ethanol not classified as dangerous goods. Based on this study's results, we recommend NaCl as a storage substrate for forensic samples in cases where no cooling/freezing conditions are available.

Blood taken immediately after fatal resuscitation attempts yields higher quality DNA for genetic studies as compared to autopsy samples.

BACKGROUND: The out-of-hospital cardiac arrest (OHCA) in the young may be associated with a genetic predisposition which is relevant even for genetic counseling of relatives. The identification of genetic variants depends on the availability of intact genomic DNA. DNA from autopsy may be not available due to low autopsy frequencies or not suitable for high-throughput DNA sequencing (NGS). The emergency medical service (EMS) plays an important role to save biomaterial for subsequent molecular autopsy. It is not known whether the DNA integrity of samples collected by the EMS is better suited for NGS than autopsy specimens. MATERIAL AND METHODS: DNA integrity was analyzed by standardized protocols. Fourteen blood samples collected by the EMS and biomaterials from autopsy were compared. We collected 172 autopsy samples from different tissues and blood with postmortem intervals of 14-168 h. For comparison, DNA integrity derived from blood stored under experimental conditions was checked against autopsy blood after different time intervals. RESULTS: DNA integrity and extraction yield were higher in EMS blood compared to any autopsy tissue. DNA stability in autopsy specimens was highly variable and had unpredictable quality. In contrast, collecting blood samples by the EMS is feasible and delivered comparably the highest DNA integrity. CONCLUSIONS: Isolation yield and DNA integrity from blood samples collected by the EMS is superior in comparison to autopsy specimens. DNA from blood samples collected by the EMS on scene is stable at room temperature or even for days at 4 °C. We conclude that the EMS personnel should always save a blood sample of young fatal OHCA cases died on scene to enable subsequent genetic analysis.

Detecting DNA damage in stored blood samples.

Several commercially available quantitative real-time PCR (qPCR) systems enable highly sensitive detection of human DNA and provide a degradation index (DI) to assess DNA quality. From routine casework in forensic genetics, it was observed that DNA degradation in forensic samples such as blood samples stored under sub-optimal conditions leads to visible effects in multiplex analyses of short tandem repeat markers (STRs) due to decreased amplification efficiencies in longer amplicons. It was further noticed that degradation indices often remain below the value that is considered to be critical. Thus, the aim of this work was to systematically analyze this effect and to compare conventional qPCR assays with a modified qPCR approach using uracil DNA glycosylase (UNG) and DNA quality assessment methods based on electrophoresis. Blood samples were stored at three different storage temperatures for up to 316 days. Significantly increased DNA recovery was observed from samples stored at high temperatures (37 °C) compared samples stored at room temperature and 4 °C. We observed typical effects of degradation in STR analyses but no correlation between DI and storage time in any of the storage conditions. Adding UNG slightly increased the sensitivity of detecting DNA degradation in one of the qPCR kits used in this study. This observation was not confirmed when using a second qPCR system. Electrophoretic systems did also not reveal significant correlations between integrity values and time. Methods for detecting DNA degradation are usually limited to the detection of DNA fragmentation, and we conclude that degradation affecting forensic STR typing is more complex.

From Deutsche Zeitschrift to International Journal of Legal Medicine-100 years of legal medicine through the lens of journal articles : Part 1: Deutsche Zeitschrift für die gesamte gerichtliche Medizin from 1922 to 1944.

From its launc h in 1922 to the end of the Second World War, the Deutsche Zeitschrift für die gesamte gerichtliche Medizin spanned 38 volumes. The 1762 papers contained in those volumes reflect contemporary interests and include many papers from peripheral fields and non-medico-legal disciplines. Publications concerned with issues outside core legal medicine fields in particular allow two distinct tendencies in the development of German institutes of legal medicine to be discerned. Firstly, there is a focus on the psychological and psychiatric aspects of the discipline. Secondly, there is tendency towards a scientific-criminalistic outlook. The fatal consequences of the Nazi seizure of power in 1933 did not spare the sciences. For legal medicine, a discipline with close links to the state, it is unsurprising that fundamental changes to the political system had a significant impact on subject matter. Leaving aside articles notable principally for their ideological content, our analysis of the 38 volumes shows that the papers examined contain new insights into many subjects, some of which are still valid today.

Fractures and skin lesions in pediatric abusive head trauma: a forensic multi-center study.

Abusive head trauma (AHT) and its most common variant, the shaken baby syndrome (SBS), are predominantly characterized by central nervous system-associated lesions. Relatively little data are available on the value of skeletal and skin injuries for the diagnosis of SBS or AHT. Thus, the present study retrospectively investigated 72 cases of living children diagnosed with the explicit diagnosis of SBS during medico-legal examinations at three German university institutes of legal medicine. The risk of circular reasoning was reduced by the presence of 15 cases with confession by perpetrators. Accordingly, the comparison with the 57 non-confession cases yielded no significant differences. Skeletal survey by conventional projection radiography, often incomplete, was found to be performed in 78% of the cases only. Fractures were found in 32% of the cases. The skull (43%) and ribs (48%) were affected most frequently; only 8% of the cases showed classic metaphyseal lesions. In 48% of the cases, healing fractures were present. Skin lesions (hematomas and abrasions) were found in 53% of the cases with the face (76%), scalp (26%), and trunk (50%) being the major sites. In 48% of the cases, healing skin lesions were observed. Nearly 80% of the cases with fractures also showed skin lesions. The data prove that SBS is frequently accompanied by other forms of physical abuse. Therefore, skeletal survey is indispensable and should always be done completely and according to existing imaging guidelines if child abuse is suspected.

Mass SARS-CoV-2 Testing in a Dormitory-Style Correctional Facility in Arkansas.

Objectives. To assess SARS-CoV-2 transmission within a correctional facility and recommend mitigation strategies.Methods. From April 29 to May 15, 2020, we established the point prevalence of COVID-19 among incarcerated persons and staff within a correctional facility in Arkansas. Participants provided respiratory specimens for SARS-CoV-2 testing and completed questionnaires on symptoms and factors associated with transmission.Results. Of 1647 incarcerated persons and 128 staff tested, 30.5% of incarcerated persons (range by housing unit = 0.0%-58.2%) and 2.3% of staff tested positive for SARS-CoV-2. Among those who tested positive and responded to symptom questions (431 incarcerated persons, 3 staff), 81.2% and 33.3% were asymptomatic, respectively. Most incarcerated persons (58.0%) reported wearing cloth face coverings 8 hours or less per day, and 63.3% reported close contact with someone other than their bunkmate.Conclusions. If testing remained limited to symptomatic individuals, fewer cases would have been detected or detection would have been delayed, allowing transmission to continue. Rapid implementation of mass testing and strict enforcement of infection prevention and control measures may be needed to mitigate spread of SARS-CoV-2 in this setting.

Sudden infant death syndrome: deletions of glutathione-S-transferase genes M1 and T1 and tobacco smoke exposure.

In developed countries, sudden infant death syndrome (SIDS) is the leading cause of death in infants in their first year of life. The risk of SIDS is increased if parents smoked during pregnancy and in presence of the child. Glutathione S-transferases (GSTs) catalyse the conjugation of glutathione with electrophilic compounds and toxins, making them less reactive and easier to excrete. As a gene dose effect was observed for GSTM1 and GSTT1, the aim of this study was to investigate whether there is a connection between homozygous or heterozygous gene deletions of GSTM1 or GSTT1 and the occurrence of SIDS. We found that heterozygous deletion of GSTM1 occurred significantly more frequently in the SIDS case group compared to the control group. A homozygous deletion of GSMT1 was slightly more frequently in the control group. A homozygous gene deletion of GSTT1 showed no significant difference between the SIDS group and the control group. We also found that in the SIDS group, the number of victims that were exposed to cigarette smoke was significantly higher than the number of victims without cigarette smoke exposure and that the mean lifetime of children whose mothers smoked was shorter in comparison with non-smoking mothers. In SIDS cases with homozygous gene deletions of GSTM1, the median life span of children with tobacco smoke exposure was 60 days shorter than without smoke exposure. In conclusion, the absence of these two genes is not the only trigger for SIDS but could be a critical aspect of SIDS aetiology, particularly in SIDS cases with smoking parents.

Abusive head trauma in court: a multi-center study on criminal proceedings in Germany.

The shaken baby syndrome (SBS) is a common variant of abusive head trauma (AHT) in infants and toddlers. Data on the legal outcome of such cases are still sparse. By means of a retrospective multi-center analysis, 72 cases of living children diagnosed with SBS/AHT from three German university institutes of legal medicine were identified. Forty-six of these cases with 68 accused individuals were available and could be evaluated with regard to basic data on the course of the criminal proceedings as well as the profile of the defendants (sub-divided into suspects, convicts, and confessed perpetrators). Criminal proceedings predominantly commenced with a complaint by the treating hospital (62%) and were found to be closed (without judgment) in 50% of the cases, mostly due to a "lack of sufficient suspicion." Of the 23 cases with judgment, the court decided on acquittal in 4 cases (17%). Imprisonment was the most frequent sentence (16 out of 19 cases with conviction, 84%), whereby the sentence has been suspended on probation in 63% of the cases. Suspects and perpetrators were mostly male and derived from the close family environment of the injured children. All confessed perpetrators stated an "excessive demand" as the reason for the violent shaking of the child. The results of the present study are in line with data from other studies with other legal systems. As many criminal proceedings were closed and the 4 acquittals occurred because the perpetration could not be ascribed to a specific perpetrator, improving the forensic methods for such an unequivocal assignment would be desirable.

Primary Indicators to Systematically Monitor COVID-19 Mitigation and Response - Kentucky, May 19-July 15, 2020.

State and local health departments in the United States are using various indicators to identify differences in rates of reported coronavirus disease 2019 (COVID-19) and severe COVID-19 outcomes, including hospitalizations and deaths. To inform mitigation efforts, on May 19, 2020, the Kentucky Department for Public Health (KDPH) implemented a reporting system to monitor five indicators of state-level COVID-19 status to assess the ability to safely reopen: 1) composite syndromic surveillance data, 2) the number of new COVID-19 cases,* 3) the number of COVID-19-associated deaths,† 4) health care capacity data, and 5) public health capacity for contact tracing (contact tracing capacity). Using standardized methods, KDPH compiles an indicator monitoring report (IMR) to provide daily analysis of these five indicators, which are combined with publicly available data into a user-friendly composite status that KDPH and local policy makers use to assess state-level COVID-19 hazard status. During May 19-July 15, 2020, Kentucky reported 12,742 COVID-19 cases, and 299 COVID-19-related deaths (1). The mean composite state-level hazard status during May 19-July 15 was 2.5 (fair to moderate). IMR review led to county-level hotspot identification (identification of counties meeting criteria for temporal increases in number of cases and incidence) and facilitated collaboration among KDPH and local authorities on decisions regarding mitigation efforts. Kentucky's IMR might easily be adopted by state and local health departments in other jurisdictions to guide decision-making for COVID-19 mitigation, response, and reopening.

Updating the risk profile of fatal head trauma: an autopsy study with focus on age- and sex-dependent differences.

Fatal head trauma (FHT) represents one of the most frequent causes of death diagnosed in forensic pathology. However, profound statistic autopsy data on FHT is still sparse. Therefore, the purpose of this study was to investigate the circumstances and injury patterns of FHT with particular focus on age and sex, and additionally, to describe a recent risk profile of FHT. To this end, the forensic autopsy records of each FHT case at a large German university hospital during a 10-year period (2006-2015) were analyzed retrospectively (n = 372). The male-female ratio was 2.6:1. Regarding median age, females were 12.5 years older than males. Traffic-associated FHT represents the major mechanism of death, followed by fall-associated FHT. While accident was the major manner of death and presented a similar distribution of age and sex, homicides were the only subgroup with a significantly lower ratio between males and females. Skull fractures occurred in 78.2% and intracranial hemorrhages in 80.6% of all cases. In summary and partly in contrast to clinical data on head trauma, FHT still occurs predominantly in male individuals under the age of 45 years, in the context of traffic accidents and affected by alcohol intake. Improvements in traffic security as well as continuing surveillance of the incidence of FHT by forensic autopsies are necessary to further reduce the incidence of FHT.

Dental age estimation based on the radiographic visibility of the periodontal ligament in the lower third molars: application of a new stage classification.

The purpose of the present study was to test whether a new stage classification based on radiographic visibility of the periodontal ligament in lower third molars in a Chinese population can be used for the 18- and 21-year thresholds. A total of 1300 orthopantomograms, including equal numbers of northern Chinese males and females evenly distributed between the ages of 15 and 40 years, were analyzed. The stages were defined according to the visibility of periodontal ligament for the outer parts of lower third molar roots because the visibility status of the periodontal ligament between the roots of lower third molars is none valuable in many Chinese individuals. Stage 0 was first achieved at the age of 17.05 years in males and 17.46 years in females. The earliest appearance of stage 1 was 17.47 years in males and 17.86 years in females. Stage 2 was first observed in males at the age of 21.43 years and in females at the age of 21.96 years. The onset of stage 3 was first observed at the age of 25.83 years in males and 23.14 years in females. Compared with the stage classification of Olze et al., which also considers the mesial parts of the roots, the number of assessable cases could be significantly increased. Therefore, our novel approach is effective for age estimation in the Chinese population.

Multiplex analysis of genetic polymorphisms within UGT1A9, a gene involved in phase II of Δ9-THC metabolism.

We present a novel multiplex assay for the simultaneous detection of 12 polymorphisms within the UGT1A9 sequence, which codes for enzymes involved in phase II biotransformation. The assay combines a multiplexed amplification step with single-base extension sequencing. The method described here is fast, cost-effective, and easy-to-use, combining the relevant features of screening methods for research and diagnostics in pharmacogenetics. To validate the assay, we tested reproducibility and sensitivity and analysed allele frequencies of 110 Caucasian individuals. Furthermore, we describe combining genetic information of individuals consuming Cannabis sativa products with respective plasma concentrations of a metabolite.

Age assessment in the living using modified Gustafson's criteria in a northern Chinese population.

Regressive dental changes appear to be suitable for age assessment in living adults. In 2012, Olze et al. showed that several criteria presented by Gustafson for extracted teeth can also be applied to orthopantomograms. The objective of this study was to test the applicability and reliability of this method in a Chinese population. For this purpose, 1300 orthopantomograms of 650 female and 650 male Chinese aged between 15 and 40 years were evaluated. The characteristics of secondary dentin formation, periodontal recession, attrition, and cementum apposition were reviewed in all the mandibular premolars. The sample was split into a training and test dataset. Based on the training set, the correlation of the individual characteristics with chronological age was studied with a stepwise multiple regression analysis, in which individual characteristics formed the independent variable. According to the results, the R values amounted to 0.80 to 0.83; the standard error of estimate was 4.29 to 4.75 years. By analyzing the test dataset, the accuracy of the present study, Olze's and Timme's formulas were determined by the difference between the estimated dental age (DA) and chronological age (CA). Taking both mean differences and mean absolute differences into account, the Chinese age estimation formula did not always perform better compared with Olze's and Timme's formulas for both males and females. It was concluded that this method can be used in Chinese individuals for age assessment. However, the applicability of the method is limited by the quality of the X-ray images, and the method should only be applied by experienced forensic odontologists.

Forensic age assessment by 3.0T MRI of the knee: proposal of a new MRI classification of ossification stages.

PURPOSE: To explore the possibility of determining majority via a morphology-based examination of the epiphyseal-diaphyseal fusion by 3.0 T magnetic resonance imaging (MRI), a prospective cross-sectional study developing and applying a new stage classification was conducted. MATERIALS AND METHODS: 344 male and 350 female volunteers of German nationality between the ages of 12-24 years were scanned between May 2013 and June 2015. A 3.0 T MRI scanner was used, acquiring a T1-weighted (T1-w) turbo spin-echo sequence (TSE) and a T2-weighted (T2-w) TSE sequence with fat suppression by spectral pre-saturation with inversion recovery (SPIR). The gathered information was sifted and a five-stage classification was formulated as a hypothesis. The images were then assessed using this classification. The relevant statistics were defined, the intra- and interobserver agreements were determined, and the differences between the sexes were analysed. RESULTS: The application of the new classification made it possible to correctly assess majority in both sexes by the examination of the epiphyses of the knee joint. The intra- and interobserver agreement levels were very good (κ > 0.80). The Mann-Whitney-U Test implied significant sex-related differences for most stages. CONCLUSION: Applying the presented MRI classification, it is possible to determine the completion of the 18th year of life in either sex by 3.0 T MRI of the knee joint. KEY POINTS: • Based on prospective referential data a new MRI classification was formulated. • The setting allows assessment of the age of an individual's skeletal development. • The classification scheme allows the reliable determination of majority in both sexes. • The staging shows a high reproducibility for instructed and trained professional personnel. • The proposed classification is likely to be adaptable to other long bone epiphyses.

8ß-OH-THC and 8ß,11-diOH-THC-minor metabolites with major informative value?

The ∆9-tetrahydrocannabinol (THC) metabolites 8ß-hydroxy-THC and 8ß,11-dihydroxy-THC are mentioned in the literature as potential blood markers of recent cannabis use. However, the formation of these metabolites in in vivo detectable concentrations has been described controversially. Therefore, the aim of this study was to verify the in vivo metabolism of 8ß-hydroxy-THC and 8ß,11-dihydroxy-THC in order to evaluate their potential as blood markers of recent cannabis use. First, we developed and validated a solid-phase-extraction method coupled with gas chromatography-mass spectrometry in order to enable the selective and very sensitive determination of 8ß-hydroxy-THC and 8ß,11-dihydroxy-THC. The application of this method in the analysis of 70 authentic plasma samples of cannabis users revealed positive results for both analytes. We detected 8ß-hydroxy-THC in three and 8ß,11-dihydroxy-THC in 37 out of the 70 analyzed samples. For 8ß-hydroxy-THC, all of the three positive results were below the limit of quantification (LOQ; 0.3 ng/mL) but above the limit of detection (LOD; 0.2 ng/mL). For 8ß,11-dihydroxy-THC, only two positive results were below the LOQ (0.4 ng/mL) but above the LOD (0.3 ng/mL); the remaining 35 were quantified. Hence, we were able to prove the in vivo metabolism from THC to both 8ß-hydroxy-THC and 8ß,11-dihydroxy-THC in detectable concentrations. The quantitative comparison of 8ß-hydroxy-THC and 8ß,11-dihydroxy-THC with the main cannabinoids THC, 11-hydroxy-THC, and 11-nor-9-carboxy-THC revealed no further informative value for 8ß-hydroxy-THC regarding the last time of cannabis consumption. However, the detectability from 8ß,11-dihydroxy-THC compared to 11-hydroxy-THC suggests a shorter detection time for 8ß,11-dihydroxy-THC and thereby a promising application of this metabolite as a blood marker of recent cannabis use.

Marker evaluation for differentiation of blood and menstrual fluid by methylation-sensitive SNaPshot analysis.

The differentiation of blood and menstrual fluid is especially important in cases of alleged sexual assault. While the identification of blood is relatively straightforward, the identification of menstrual fluid in trace evidence has been shown to be more challenging. This may be due to the complex nature of menstrual fluid that leads to intra- and inter-individual differences in composition. Nevertheless, recent advances in DNA methylation profiling have revealed promising markers for the differentiation of the two body fluids and furthermore, markers to distinguish menstrual fluid from vaginal fluid. A literature study was performed and in total, 11 markers were evaluated in this study of which seven could be validated for menstrual fluid and blood identification purposes. Marker "BLU2" (chr16:29757334) was identified as most suitable for differentiation of blood and menstrual fluid.