Quantitative analysis of amplifiable DNA in tissues exposed to various environments using competitive PCR assays

Competitive PCR assays were established for the mitochondrial DNA hypervariable region I and the human amelogenin locus. Using these assays, the copy numbers of DNA participating in PCR (amplifiable DNA) were quantified in tissues exposed to different environments. Human ribs, skin and nails were left in three exposure conditions (in the open air, in soil and in water). The amounts of amplifiable DNA in these tissues were quantified during a time period of up to two months. The amount of amplifiable DNA was well preserved in hard tissues (ribs and nails) regardless of the exposure conditions, whereas the soft tissues immersed in water showed a rapid decrease in amplifiable DNA. Strong PCR inhibition was observed in the DNA extracts obtained from buried bones. This phenomenon was clearly identified from an amplification failure of the internal standards in the competitive PCR. A preliminary examination to identify the PCR inhibitor suggested that the soil itself contributed to the inhibition. In addition, the amounts of amplifiable DNA in case samples were also investigated.     

指印生物遗传标记检验的研究现状与展望

综述了血指印和汗潜指印生物遗传标记的研究历史和现状,指印DNA检验面临的问题,包括现场污染,显现剂的影响,载体的影响等,并对指印DNA在法医学中的应用前景进行了展望。     

微量生物检材常规初检后再行PCR检验的方法

在实际检案中，经常遇到案发现场可获取的生物检材量微，在进行必要的种属检验、精斑确证实验、ABO血型检验等常规物证初检后，便无多余的生物检材移送DNA实验室进一步做法医DNA检验。因此，笔者通过对检案中遇到的上述微量物证检材的再行处理利用，在本实验室条件下，对其进行了TH０１、HUMACTBP２、AluVpA、DIS８０等位点的DNA－PCR分析，获得了良好的效果。使其在实际办案中更充分地发挥了证据作用，报告如下。１材料与方法检案中已经种属或ABO血型检验后的血痕、唾液斑（如烟蒂外层纸）浸泡凹板，加入３００μl去离子水，…     

纸张上潜在手印三种前处理方法对DNA检验的影响

目的通过比较常见纸张上潜在手印盲提法与显现后精准提取法的接触DNA检出率,探讨常见纸张上接触DNA前处理的优选方案。方法比较五种常见纸张上使用盲提法和显现潜在手印(茚二酮显现法、茚三酮熏显法)后精准提取法采集的接触DNA样本检出率。结果粗糙日历纸盲提的接触DNA检出率为17.8%,通过茚二酮法和茚三酮法显现的潜在手印所提取的DNA检出率分别为75.6%、77.8%;光滑日历纸三种方法所提取的接触DNA检出率为4.4%、11.1%、11.1%;A4复印纸三种方法接触DNA检出率为20%、37.8%、66.7%;牛皮纸三种方法接触DNA检出率为20%、68.9%、64.4%;快递纸袋的三种方法接触DNA检出率为2.2%、6.7%、46.7%。结论不同纸张上潜在手印经显现后接触DNA检出率不同,通过茚二酮或茚三酮显示潜在手印后精准提取DNA的前处理方法相较于盲提法的接触DNA检出率高。实战中可应用此类方法同时获得手印与DNA分型,以有效提高证据力。     

The effectiveness of forensic evidence in the investigation of volume crime scenes

This study investigates the effectiveness of forensic evidence in UK volume crime investigations. The main aim was to identify characteristics of forensic evidence that influence its effectiveness in converting detections into criminal charges, as well as to critically consider the effectiveness of a recent service level agreement (SLA) implemented by Wiltshire Police, which aimed at reducing CSI attendance. The sample consisted of 445 police recorded cases received from Wiltshire Police. Presence or absence and location-related characteristics of fingerprint, DNA, and footwear evidence were evaluated on the effectiveness of forensic evidence and examined within the contexts of different volume crimes. Results showed a high level of correlation in converting detections into criminal charges where the presence of DNA, footwear, and multiple evidence types was recorded; and a positive correlation between forensic evidence ineffectiveness and presence of fingerprints, particularly in residential burglaries. Differences between individual offence types were expressed. The most prominent feature influencing the effectiveness of forensic evidence was found to be related to the movability of the exhibit associated with the recovered evidence, with DNA recovered from non-movable items presenting the strongest effectiveness. Cases processed after the implementation of the SLA did not show significant differences in forensic evidence effectiveness as compared to cases processed prior to the SLA, however, they demonstrated a lack in effectiveness of DNA evidence. The findings of the current research provide a better understanding of the contextual influences on the potential of forensic evidence and can support improvement of crime scene screening and CSI resource deployment.     

DNA-based and taxonomic identification of forensically important Sarcophagidae (Diptera) in southeastern Spain

Studying dipterans at the scene of a death can provide essential information for interpreting the evidence and help to reconstruct the events happened to a corpse in the past. Molecular tools have been employed for identification at specific levels in the cases of cryptic species or poorly conserved specimens. Identification of specimens is essential in forensic entomology since each species has a specific growth rate, which determines the calculation of the minimum post mortem interval (minPMI). In addition, phylogeographic reconstruction within a species can help to differentiate the haplotypes from a geographic area, thereby helping to clarify the possible relocation of a corpse. The morphological identification of Sarcophagidae species is often difficult, especially for the females. This is an important Diptera family since some of its species are among the first to reach a corpse, especially in warm areas. In this study, we compared the sarcophagids found in human corpses in forensic cases in Alicante (southeast of Spain) with specimens collected from baited traps in the same area and surrounding provinces. In total, 189 specimens were collected, comprising 72 from forensic cases and 117 from baited traps. Molecular identification was conducted by sequencing the cox1 mitochondrial gene and analyzing the sequences using ABGD, GMYC, and BIN species delimitation methods. The median joining algorithm in the PopART program was used to construct phylogeographic networks. Eight species in the family Sarcophagidae were identified. The most widely collected species were Sarcophaga argyrostoma and Sarcophaga tibialis. The haplotype networks obtained for these species did not indicate a clear geographic distribution of haplotypes. The S. argyrostoma samples from Alcoy were clearly isolated. The results demonstrated that this method is useful for identifying Sarcophagidae samples in forensic investigations and it can be employed for minPMI estimation.     

Methods for ensuring the highest DNA concentration and yield in future and retrospective trace DNA extracts

In forensic laboratories, increased extraction efficiency of trace evidence is paramount because analytical success is intrinsically dependent on the quantity of DNA recovered. Moreover, highly concentrated nucleic acids are vital for effective downstream analysis and high quality results. This study investigated the efficiency of extraction with the Qiagen® QIAamp® DNA Investigator kit, and explored improvements to the methodology that would maximise the recovery of low concentration forensic samples. Controlled amounts of starting cellular material were used to mimic trace (or low level) DNA deposits prior to DNA extraction with the Investigator kit. Addition of the provided carrier RNA along with conducting two successive elutions of 50 µL improved the net recovery of DNA to 95%. Concentration with centrifugal filters post-extraction were able to concentrate DNA but a large net loss was observed. For the concentration of historic, retrospectively extracted DNA, centrifugal methods are able to concentrate DNA extracts previously too dilute for analysis. These concentrated volumes, however are small, allowing for minimal downstream analysis attempts before the sample is exhausted.     

Blood and sperm traces on human hair. A study on preservation and detection after 3-month outdoor exposure

Hair can retain important biological traces for forensic investigations. Forensic scientists are used to looking for such traces on clothing and skin of victims, however, when decomposition kicks in and all that remains of the victims is the skeleton, hair may be the only tissue representing the surface of the body at the time of a crime on which biological traces of an aggressor may have been left and still be detectable. Given the lack of research on this topic, this pilot study aims to assess the capacity of hair to retain semen and blood in hair, and the possibility to detect these fluids with well-known techniques and to obtain a useful genetic profile even when exposed to environmental conditions (Open Natural Environment (woods), Open Man Made Environment (urban)) for three months.Results showed that both traces were always visible and detectable with almost all techniques in the Control Environment, while in the two open environments some difficulties arose. However, biomolecular analysis was effective up to three months on both fluids in the Natural Environment and up to two months and one week respectively on blood and semen in the Man Made Environment. The Combur Test, OBTI, and Luminol were effective on blood up to three months in both environments while Sperm-HY-Liter and observation of cellular components were effective on semen up to at least 1 month and PSA testing was positive up to 1 week in both environments.The present work can be considered an encouraging starting point for the analysis of biological traces on hair in forensic contexts, regardless of the PMI, since blood and semen related to a crime may survive.     

基于年龄相关DNA甲基化位点推断河南汉族个体年龄

目的探查中国河南汉族个体与年龄相关的DNA甲基化位点,构建年龄推断模型,进行甲基化和年龄相关性分析。方法采用焦磷酸测序法对ELOVL2、ClOrf132、KLF14、TRIM59和FHL2基因的34个CG位点进行甲基化分析,利用SPSS 23软件的多元回归方法建立模型,对甲基化和年龄相关性做分析。结果除ClOrf132基因3个CG位点的甲基化水平与年龄呈负相关外,其余4个基因的31个CG位点甲基化水平均与年龄呈正相关。多元回归分析表明,年龄与CG位点的甲基化水平存在明显的线性关系,实际年龄与推断年龄偏差在5岁以内的准确度达80%以上。结论本研究构建的河南汉族个体年龄推断模型,有助于通过检测血液等组织的DNA甲基化水平推断个体的年龄范围,具有法医学应用前景。     

Nondestructive Methods for Recovery of Biological Material from Human Teeth for DNA Extraction

The extraction of DNA from human skeletal remains applied to forensic, and evolutionary studies do not exclude risks, which are to be evaluated when working with unique specimens that could be damaged or even destroyed. In the present study were evaluated several nondestructive methods for recovering DNA instead of the most currently used pulverization method. Three different procedures to access inside the dental pieces (occlusal perforation, cervical perforation, and cervical cut) have been compared with the aim of recovering as many cell remains as possible to carry out a DNA extraction. Given the DNA quantitation results, a method was proposed that consists of a cervical cut to facilitate the access to the pulp cavity and a subsequent filing of the root canals down to the apex of the dental root. This methodology allows the recovery of both mitochondrial and nuclear DNA, with the minimum deterioration for the dental pieces.