现场DNA生物检材的提取及注意事项

刑事案件的现场一般会遗留有受害人及嫌疑人的DNA生物检材,要成功检验这些DNA检材并将其转化为证据,如何发现、提取DNA检材就显得至关重要.刑事案件中适合DNA检验的生物检材包括血液(痕)、精液(斑)、唾液(斑)、脱落上皮细胞、毛发、组织、骨骼等.     

Detection of cellular material within handprints

A novel technique for the visualisation of cellular material has been published harnessing an external binding nucleic acid fluorescence dye, Diamond™ dye (DD), in combination with a digital fluorescence microscope. This technique can effectively detect cellular material on an object transferred by touch allowing targeted collection of latent DNA. Previous studies on the visualisation of touch DNA have focussed on transfer from fingertips only.Here we report on the visualisation of cellular material transferred via twenty different positions over the entire handprint. Three volunteers (a heavy, an intermediate and a light shedder) were asked to press their hands onto a plastic surface with medium pressure for 15 s at undefined time points post-handwashing, creating a complete handprint. DD was applied to the entire area and the presence of cellular material was recorded based on cells within 5 separate frames at each of the 20 positions. All tests were performed in triplicate such that the final dataset contained 1,800 observed frames.This extensive study allows accurate monitoring of cellular transfer deposited by different parts of the hand. Our study highlights which areas of an individual’s hand shed the greatest, or least, amount of cellular material. This simple process can act as a guide for DNA collection from items held within the entire hand, rather than only touched by the fingertips only, such as weapons, knives and steering wheels.     

Species identification in routine casework samples using the SPInDel kit

The identification of species in casework samples is of fundamental importance for forensic investigations. Laboratories are increasingly compelled to provide accurate and fast identifications in trace materials left on crime scenes, wildlife poaching, illegal trade of protected species, fraudulent food products cases, etc. However, the field of nonhuman forensic genetics is still working on the standardization of typing methods and practices. Here we describe the successful implementation of the Species Identification by Insertions/Deletions (SPInDel) method in routine casework analyses in 11 laboratories worldwide. The SPInDel was developed to detect human DNA, at the same time that identifies common animal species. The fragment size analysis of six mtDNA regions allows identification in suboptimal DNA samples, including mixtures, with no need for sequencing. The samples were collected from 2013 to 2018 and included hair, blood, meat, saliva, faeces, bones, etc. The SPInDel kit successfully identified >95% of the samples, being dog, human and pig the most frequently detected species. The six SPInDel loci were successfully amplified in mixtures and degraded samples (river water, sand, stains in clothes, etc.). Interestingly, several species that were not originally targeted by SPInDel primers were also identified (e.g., red fox, brown bear, fallow deer and red deer). In conclusion, the SPInDel kit was successfully used in crime scene investigations (often involving human DNA detection) and in cases of poaching, environmental contamination and food fraud. It is now becoming a useful tool for the routine analysis of nonhuman DNA samples within the high quality standards of forensic genetics.     

Evaluation of 19 short tandem repeat markers for individualization of Papaver somniferum

Papaver somniferum, commonly known as opium poppy, is the source of natural opiates, which are used as analgesics or as precursors in the creation of semi-synthetic opioids such as heroin. An increase in opioid addiction in the United States has resulted in high rates of illicit opioid use and overdoses. It has recently been shown that P. somniferum DNA suitable for genetic analysis can be recovered from heroin samples. The development of a comprehensive genetic individualization tool for opium poppy could serve to link cases and strengthen programs such as the Drug Enforcement Administration’s (DEA) Heroin Signature Program, which seeks to combat rising opioid use.The purpose of this study was to develop a quantitative real-time PCR (qPCR) method for the quantification of opium poppy DNA, compare three commercial DNA extraction kits for their ability to isolate DNA from poppy seeds, and evaluate nineteen opium poppy short tandem repeat (STR) markers for their use in a forensic identification panel. Such a panel could be used for individualizing samples and determining the geographic origin in heroin or poppy seed tea cases. The qPCR method was proven to be reproducible and reliable, specific for P. somniferum, and sensitive enough for forensic case-type samples. Of the three kits tested, the nexttec™ one-step DNA Isolation Kit for Plants was the optimal method and facilitated rapid extraction of DNA from poppy seeds. The majority of evaluated STR primer sets were unreliable or had low discriminatory power, limiting their use for individualization of poppy samples. A six-locus STR multiplex was developed and evaluated according to Scientific Working Group on DNA Analysis Methods (SWGDAM) and International Society of Forensic Genetics (ISFG) guidelines, including the use of a sequenced allelic ladder. The multiplex was found to have low discriminatory power, with greater than two-thirds of samples analyzed having just two different genotypes. The multiplex was determined to be unsuitable for individualization; however, a genotype map was developed as a proof of concept that these markers may be useful for determining the biogeographical origin of samples. Searching the poppy genome for new STR markers and developing new primer sets may be necessary for the creation of a powerful genetic tool for the individualization of P. somniferum.     

Evaluating the Efficiency of Primer Extension Capture as a Method to Enrich DNA Extractions,

In this study, we sought to document the efficiency of primer extension capture (PEC) as a method to enrich DNA eluates of targeted DNA molecules and remove nontarget molecules from pools containing both. Efficiency of the method was estimated by comparing number of “copies in” to “copies out” by quantitative polymerase chain reaction. PEC retention of DNA targets ranging 109‐288 base pairs (bps) in length was 15.88–2.14% (i.e., loss of 84.12–97.86% of target molecules). Experimental modifications of the PEC method resulted in no significant improvements. However, the benefit of PEC was revealed in its ability to remove most nontarget DNA molecules (99.99%). We also discovered that many (56.69%) of the target molecules are “lost” prior to their immobilization on the streptavidin‐coated beads. These estimates of methodological efficiency are directly comparable to previous ones observed following “fishing” for DNA, an alternative method for DNA enrichment.     

Forensic DNA retention: Public perspective studies in the United Kingdom and around the world

This review analysed public perspective studies on forensic DNA retention in the United Kingdom and around the world. The studies generally show strong public support for the long-term or indefinite retention of DNA from convicts and suspects. There is considerable support for the retention of DNA from all or some arrestees and potentially the entire population. This was predicated upon the belief that forensic DNA databases have crime-solving abilities, which the public rate highly. In the UK, it was found that the current Protection of Freedoms Act 2012 regime is broadly representative of the recommendations of the surveyed British public. Nevertheless, the studies highlighted a gap in forensic DNA education among the public, suggesting that public views may not be well informed. Overall, there was clear evidence of privacy concerns and the potential misuse of DNA records among the public, with a significant number opposing the retention of DNA from the innocent. It was found that most of the studies were qualitative or non-representative of the relevant population, limiting the generalisation of the results. There was also limited studies among a representative sample of primary stakeholders who are well-informed or directly exposed to the benefits, challenges and risks associated with DNA retention. A research into stakeholders rating of different forensic DNA retention regimes is therefore highly recommended. This is important because the studies suggest divergent views among criminal justice professionals and other members of the public, with the former expressing expansive views and the latter expressing restrictive views. The primary stakeholder's survey will help establish whether the relevant safeguards have been put in place to protect both public security and individual interests.     

纳米磁珠提取法在骨骼DNA提取中的应用

目的对纳米磁珠法提取纯化骨骼DNA的效果进行比较评价,为方法选择提供应用参考。方法取泥土掩埋、水中浸泡1~10年不等的25根长骨,经水洗、刮净,液氮冷冻研磨器将骨骼研磨成粉末状,分别应用纳米磁珠提取法和King Fisher仪器自动化提取法提取DNA,IdentifilerPlus试剂盒进行扩增,ABI 3100遗传分析仪进行STR分型检测;对两种方法提取的DNA定量和经扩增、分型检测的结果进行比较。结果骨骼样本采用纳米磁珠法提取到的样本DNA(1.237 5ng/μL±0.319 2ng/μL),较之King Fisher法的浓度(0.506 2ng/μL±0.280 5ng/μL)更高,两种方法间差异具有统计学意义(P0.05);而纳米磁珠法的分型成功率亦更高,两种方法间差异具有统计学意义(P0.05)。结论用纳米磁珠提取纯化骨骼DNA,能得到高质量DNA模板,有利于提高分型检验的成功率,可在实际检案中选择使用。     

Distribution of Y chromosomal STRs loci in Mayan and Mestizo populations from Guatemala

In this study, a sample of 225 Guatemalan males, comprising 115 Mestizo-Guatemalan and 110 Mayan-Guatemalan, was typed for 17 Y-short tandem repeats (STRs) loci (DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438, DYS439, DYS448, DYS456, DYS458, DYS635, YGATA_H4.1 and DYS385a/b). The haplotype diversity (H = 1) and discrimination capacity (96.86%) were calculated. Analysis of molecular variance (AMOVA) demonstrated a low but significant interpopulation differentiation when compared with the results obtained when we confront the Mestizo and Mayan populations with the European populations.Furthermore, the genetic variability and differences among the American, African, Asian, and European populations were analyzed with the software Statistica 9.1. In addition, the genetic distances were also calculated using other published data. Reynolds and Slatkińs genetic distance was visualized using the multidimensional scaling (MDS) analysis. All the analysis performed locates the Mayan population next to the Native American population, while Guatemalan-Mestizo population was found to be between these populations and the European population, similar to other Mestizo one.The implementation of the estimation of individual ancestry proportions of the whole population sample showed the presence of two well-differentiated population groups.     

Belgian canine population and purebred study for forensics by improved mitochondrial DNA sequencing

In canine population studies for forensics, the mitochondrial DNA is profiled by sequencing the two hyper variable regions, HV1 and HV2 of the control region.In a first effort to create a Belgian population database some samples showed partially poor sequence quality. We demonstrated that a nuclear pseudogene was co-amplified with the mtDNA control region. Using a new combination of primers this adverse result was no longer observed and sequencing quality was improved. All former samples with poor sequence data were reanalyzed. Furthermore, the forensic canine population study was extended to 208 breed and mixed dogs. In total, 58 haplotypes were identified, resulting in an exclusion capacity of 0.92. The profile distribution of the Belgian population sample was not significantly different from those observed in population studies of three other countries.In addition to the total population study 107 Belgian registered pedigree dogs of six breeds were profiled. Per breed, the obtained haplotypes were supplemented with those from population and purebred studies. The combined data revealed that some haplotypes were more or less prominent present in particular dog breeds. The statistically significant differences in haplotype distribution between breeds and population sample can have consequences on mtDNA databasing and matching probabilities in forensics.     

Attitudes towards police use of consumer/private DNA databases in investigations

Consumer DNA products, such as databases that allow the public to explore familial relationships, have recently been used by police to assist in investigations. This has moved the collection of DNA used in criminal investigations away from the limitations of criminal databases and has opened up ethical concerns regarding privacy and consent. This study explored public attitudes and support towards police use of consumer DNA databases in investigations to assess whether different crime types or moral and attitude alignments influenced the level of public support of police using these consumer products. An international survey of 438 adults, using theory and scales pertaining specifically to retributive punishment and attitudes towards law, courts and police, found that moral alignment and attitudes did influence the level of public acceptability towards police usage of these techniques and that support did vary based on crime type. This research found that higher levels of public support was given for the most serious case types explored (sexual assault and homicide). Participant support for police access to private DNA databases by case type was as follows; 83.5% for sexual assault, 83.2% for homicide, 85.2% for identifying unidentified human remains, 62.8% for robbery and 58.9% for illicit drug related cases. Small to medium effects sizes were observed for these results. Although these findings provided context towards public attitudes, further research specific to international attitude differences and variance between serious crime types and public acceptability could further develop these findings.