DNA profiling of trace DNA recovered from bedding

Trace DNA is often detected on handled items and worn clothing examined in forensic laboratories. In this study, the potential transfer of trace DNA to bedding by normal contact, when an individual sleeps in a bed, is examined. Volunteers slept one night on a new, lower bed sheet in their own bed and one night in a bed foreign to them. Samples from the sheets were collected and analysed by DNA profiling. The results indicate that the DNA profile of an individual can be obtained from bedding after one night of sleeping in a bed. The DNA profile of the owner of the bed could also be detected in the foreign bed experiments. Since mixed DNA profiles can be obtained from trace DNA on bedding, caution should be exercised when drawing conclusions from DNA profiling results obtained from such samples. This transfer may have important repercussions in sexual assault investigations.     

DNA profiling of trace evidence--mitigating evidence in a dog biting case

A young girl was the victim of a severe dog attack. An animal, suspected of having caused the attack, was later impounded for investigation. Microclots of blood, recovered from the dog's fur, were analyzed by STR DNA. Results showed that this blood was not related to the biting. Other forensic evidence--hairs, fibers, and odontology--failed to connect a particular animal to the attack. The implications of these findings for the dog and its owners are discussed as well as other forensic methods for resolving such cases.     

鞋内底不同部位接触DNA提取初探

目的探讨鞋内底不同部位接触DNA提取检出率。方法取100名20-30岁的志愿者，将其穿用过的运动鞋和皮鞋鞋垫设置成不同穿用时间组、不同材料鞋垫组、穿用后不同放置时间组，根据脚的形态学及运动力学特点，将鞋垫分成8个区域进行脱落细胞提取，并进行DNA进行检验。结果鞋垫上8个不同区域提取到的脱落细胞DNA分型检验效果不同。足弓外侧区（足引折弓除外）的接触DNA检出率最高；足弓内侧、第1趾骨区、第1跖骨区及足跟区次之；第2～5趾骨区、第2～3跖骨区、第4～5跖骨区不容易成功提取到接触DNA。结论鞋内底接触DNA检出率与接触时间、放置时间、鞋垫材质均相关，分区提取检验DNA更有针对性。     

改良IPEP法用于痕量DNA检测的实验研究

目的探讨改良扩增前引物延伸(IPEP)法对痕量DNA样本STR检测分型的效果。方法用改良IPEP法对痕量样品DNA进行全基因组扩增(WGA),扩增产物用实时荧光定量PCR技术定量、用AmpFLSTR~ Indentifiler~试剂盒作基因型检测。结果该方法可增加模板DNA约200~1100倍。基因组DNA不低于0.025ng时,可获得15个STR基因座和Amelogenin性别基因座的分型结果。基因组DNA0.01~0.025ng时,可获得9个以上基因座的分型结果。结论改良IPEP法可有效提高痕量DNA样本STR分型检验的灵敏度,有较好的实用价值。     

Isolation of the DNA minisatellite probe MZ 1.3 and its application to DNA 'fingerprinting' analysis

A minisatellite probe, MZ 1.3, detecting hypervariable fragment patterns was isolated from a human genomic library. A repetitive sequence of 27 bp length was identified which is contained in the probe approx. 40 times. The MZ 1.3 repeat shows variable homology of 53-73% to the repetitive sequence of the protein III gene of the bacteriophage M13 genome. Polymorphic restriction fragment patterns were found with MZ 1.3 using the enzymes Hinf I, BstN I, Hae III, Mbo I, PstI/Pvu II, and Rsa I. An average of 18 polymorphic fragments was observed using Hinf I as enzyme. The band sharing frequency after Hinf I digestion among unrelated individuals was determined to be 23.8 +/- 7.2%. An example for the application of MZ 1.3 to paternity testing in an incest case is given. The probe can be used with radioactive or non-radioactive detection systems. An approach is presented to compare polymorphic fragment patterns from individuals obtained by independent gel runs on the basis of relative band positions (RBP) and calculated in a computerized analysis.     

全基因组扩增在微量检材DNA分型中的应用

目的探索全基因组扩增技术对微量检材DNA分型的有效性。方法通过显微操作制备含1~20个细胞的模拟微量检材样本,在常规PCR-STR分型前加入全基因组扩增步骤,从等位基因不平衡、等位基因丢失、基因座丢失、伪等位基因(包含stutter峰)等方面探究PEP和MDA两种全基因组扩增方法对微量检材DNA分型的有效性。结果 MDA扩增效率高于PEP,但等位基因丢失和伪等位基因严重;PEP方法的正确分型率高于MDA,但小片段DNA优势扩增现象较严重。结论 MDA方法并不适合目前以STR分型为主导的法庭科学,当微量检材样本的绝对量相当少时,可以考虑使用PEP方法来扩大样本量,以满足重复检验的要求,但可能面临大片段DNA扩增失败的风险。     

Successful extraction of human genomic DNA from serum and its application to forensic identification

We report here on the successful extraction of human genomic DNA from a serum sample in a forensic case. The extracted DNA was successfully used for the identification of remains presumably immersed for more than three weeks for which the only comparison sample was a 250-microL serum aliquot kept frozen in a laboratory. The analysis made it possible to identify a second victim as the daughter of the first.     

Toulmin's philosophy of argument and its relevance to offender profiling

This study sought to identify the extent to which claims about the probable characteristics of offenders in ‘offender profiles’ were based on substantive arguments. Because Toulmin's (1958) philosophy of argument has been demonstrated as a useful way of breaking down arguments into their constituent parts (Burleson, 1979) we examined the extent to which profiles contained grounds, warrants, backing and rebuttals to support or refute various claims about offenders. Twenty-one profiles, representing a range of ‘profiling styles’, were obtained from a variety of sources. All of these had been used in major criminal investigations either in the UK or internationally. Of the nearly 4,000 claims made, nearly 80% were unsubstantiated. That is, they contained no grounds, warrant, backing or rebuttal. Moreover, less than 31% of the claims were falsifiable. We argue that (a) this demonstrates the need for a careful, systematic evaluation of profiling advice (b) Toulmin's structure is one useful method for evaluating such material and for providing a possible framework for such advice.     

M48磁珠法和Chelex-100法提取脱落细胞DNA的初步比较

目的对M48磁珠法和Chelex-100法提取脱落细胞DNA的检验效果进行比较,为优化提取方法提供参考。方法选取案件受理检材50例烟蒂、50例纺织物品、50例作案工具,根据不同条件分别用吸附法、沾附法获得DNA,采用磁珠法（M48）和Chelex-100法提取后,常规STR检测。结果烟蒂类检材采用2种方法提取DNA,所得结果没有明显差异;纺织物品和作案工具上脱落细胞DNA的提取采用M48磁珠法提取的效果明显优于Chelex-100法。结论相对而言,M48磁珠法更具优势。     

DNA甲基化及其与个体年龄相关性研究

近年来表观遗传学研究发现,基因组DNA甲基化总体水平随年龄增加而降低,同时部分位点的甲基化水平却随年龄增加而升高。本文重点介绍了DNA甲基化与年龄的相关性及其在年龄推断方面的研究进展,旨在为法医学个体年龄推断的研究提供一种新的思路。     

DNA extraction and STR profiling from histological slides

Formalin-fixed paraffin-embedded (FFPE) tissue blocks are commonly used in the field of pathology and forensic pathology as a source of histological slides. For postmortem kinship analysis or identification, DNA can be extracted from blocks with specialized kits. However, when an STR profile should be generated from single microscope slides, the removal of the coverslip and the limited sample size poses unique challenges. We aimed to test the effectivity of agitated xylene incubation to dissolve the mounting material to facilitate the coverslip removal. DNA extraction tests were performed on 5- to 7-year-old histological slides. Xylol was used to dissolve the mounting medium to facilitate cover slide removal, one set of samples was shaken during incubation, and the other set was left still. It was found that shaking the sample while bathed in xylol decreased the incubation time from three days to two days. Agitation not just reduced the processing time but increased the quality of acquired STR profiles: on average 30% more alleles were detected from the shaken samples compared to the still bathed ones.     

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

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.     

Assessment of body fluid identification and DNA profiling after exposure to tropical weather conditions

Despite current advances in body fluid identification, there are few studies evaluating the effect of environmental conditions. The present work assessed the detection of body fluids, blood, semen, and saliva, through lateral flow immunochromatographic (LFI) tests, exposed to tropical weather conditions over time, also evaluating the possibility of obtaining STR (short tandem repeat) profiles and identifying mitochondrial DNA (mtDNA) polymorphisms. Blood, semen, saliva samples, and mixtures of these fluids were deposited on polyester clothes and exposed to open-air tropical weather conditions for 1 month. The test versions from LFI (SERATEC®, Germany) Lab and crime scene (CS) used for the detection – one per each body fluid type – demonstrated that it is possible to identify body fluids and their mixtures up to 14 days after deposition. At 30 days, blood and semen were detected but not saliva. Full STR profiles were obtained from 14-day-old blood samples, and partial profiles were obtained from the remaining samples. It was possible to sequence mtDNA in the samples previously analyzed for STR profiling, and haplogroups could be assigned. In conclusion, this study demonstrated for the first time the possibility of body fluid identification and DNA profiling after exposure to tropical weather conditions for 1 month and also demonstrated the value of mtDNA analysis for compromised biological evidence.     

变性高效液相色谱及其在法医DNA分析中的应用

变性高效液相色谱(DHPLC)是目前检测基因突变的最佳技术平台之一。本文简要介绍了DHPLC的工作原理及主要技术特点,并对其在法医DNA分析中的应用进行综述。     

Recovery and evaluation by cytologic techniques of trace material retained on bullets

Fragments of tissue, intermediate targets, and debris related to firing are embedded in the fine striations and deforming edges of bullets. Because most of these fragments are too small to visualize and process as histologic sections, this material is usually washed away when the projectiles are cleaned following removal at autopsy. By preserving the rinsing material that results from routine cleaning of projectiles, it may be possible to evaluate adherent material from the bullet by cytologic techniques, including filter preparations, cell blocks, and smears of macroscopic tissue fragments. Bullet-wash cytology produced cellular elements, tissue fragments, and inert material from intermediate targets. Different tissue elements could be documented with a given projectile; this information could be utilized to document the path of a bullet through the body or intermediate target. This initial study suggests that low- and high-velocity projectiles produce different types of tissue debris, with much more fragmentation and scarcity of cellular components in the high-velocity rounds. Inert material, resulting from intermediate targets, such as clothing, as well as gunshot residue on the bullet or debris from the barrel could be distinguished on preparations. There was a difference in tissue representation of adherent material on the bullet; connective tissue, mesothelial coverings, and fragments from organs with higher elastic and cohesive properties were seen with much greater frequency on the filters than were loosely cohesive and friable organs such as liver and spleen. The cytologic preparations from projectile washings reflect both the path taken by the bullet and the ballistic damage to the organs. Thus, the cytologic evaluation of bullet washings may be useful in the incorporation of gunshot wound evaluation to support documentation of the trajectory of the projectile.     

Recovery of DNA and fingerprints from touched documents

This study investigated the various factors affecting DNA profiling from DNA recovered from fingerprints deposited on paper before and after fingerprint enhancement treatments. The DNeasy^® plant mini kit (QIAGEN^®) was found to improve DNA recovery from paper by over 150% compared with the QIAamp^® mini kit. A significant decrease in the amount of DNA recovered was observed following treatment with DFO and/or Ninhydrin. This decrease in yield did not have a comparably significant effect on the quality of the SGM Plus™ profiles. Furthermore, this study found that whilst certain paper types, such as newspaper, magazine and filter paper allowed for the good recovery of DNA, common office paper and white card, strongly interfered with the recovery of DNA resulting in poor quality profiles.