基于psbA-trnH条形码的毒品原植物大麻及其混伪品的鉴别

目的 利用叶绿体上的间隔区psbA-trnH条形码序列鉴别毒品原植物大麻及其混伪品,为毒品原植物大麻的鉴定提供新方法.方法 采用PCR法扩增psbA-trnH间隔序列,双向测序后运用CodonCode Aligner、MEGA5.1软件进行数据处理,构建系统聚类树(NJ树).结果 psbA-trnH条形码序列分析表明大麻种内与种间遗传距离具有较大差异,基于psbA-trnH条形码构建的NJ树可鉴别大麻及其混伪品.结论 psbA-trnH 序列可以作为鉴定毒品原植物大麻及其混伪品的候选条形码序列,为毒品原植物大麻的快速、准确鉴定提供了新方法;DNA条形码技术在犯罪案件中涉及的植物鉴定中具有极大的应用前景.     

利用DNAITS2条形码序列鉴定植物大麻和罂粟

目的分析毒品原植物大麻、罂粟及其混伪品植物r DNA的ITS2序列信息,探索鉴定大麻、罂粟及其混伪品的新方法。方法采用PCR法扩增ITS2序列,双向测序后运用Codon Code Aligner、MEGA5.1软件进行数据处理,计算种内种间K2P距离,构建系统聚类树(NJ树)。结果大麻、罂粟的种内K2P遗传距离为0,大麻、罂粟及其混伪品之间的最小K2P距离为0.187。NJ树表现出明显的单系性。结论利用DNA ITS2条形码序列有可能鉴定出毒品原植物大麻、罂粟。     

GC-MS检测新型策划毒品K3

目的基于气相色谱-质谱（GC-MS）检测结合特殊质谱库信息检索建立新型策划毒品的鉴定方法。方法未知样品用甲醇超声溶解,吸取上清液采用气相色谱-质谱（GC-MS）联用仪检测。结果测得A组分（t R=19.47min）的质谱特征碎片峰（m/z）信息为215.1（基峰）、144.9、294.1、337.1和365.1,B组分（tR=23.29min）的质谱特征碎片峰（m/z）信息为359.1（基峰）、127.1、144.0、155.0、232.1、284.1和342.0。经美国缉毒署毒品分析谱库检索获得的信息资料,鉴定为新型策划毒品＂K3＂,其主要组分为＂AKB48＂和＂AM2201＂,此类化合物具有大麻类似精神活性,归属合成大麻素。结论本方法可用于新型策划毒品组分的鉴定。     

大麻三个STR基因座的遗传多态性调查

在大麻毒品犯罪案件中经常缴获到成批未加工的大麻植物,多为同一产地种植,各产地种植大麻又多以某个或某几个品种为主.如能鉴别出这些大麻的品种,则能根据各地种植大麻的情况推断出缴获大麻的毒源地,为铲除毒品源头提供重要帮助.     

缴获疑似毒品烟叶检出新型合成大麻素1例

随着互联网的发展,全球范围内的科技交流更为快捷。与此同时,制毒技术也在互联网上蔓延。为了逃避打击处理,不法分子通过在被管制毒品分子基础上增减、变换取代基位置、改变取代基结构等方式,合成一系列与管制毒品结构相似的新型毒品,合成大麻素是其中的一类[1]。自2013年起,我国开始对合成大麻素类毒品进行管制。     

疑似毒品烟丝和烟油中检出新型合成大麻素ADB-BUTINACA

正近几年,新型合成大麻素类毒品在我国广泛传播,目前相关文献已经报道了ADB-FUBINACA、5F-MDMB-PICA、AMB-FUBINACA等诸多新型合成大麻素类毒品案件~([1-4])。截止2021年4月15日,Cayman上已列出合成大麻素类化合物794种,远超芬太尼类毒品的398种。本文利用GC/MS和LC/MS/MS在山东省缴获的疑似毒品烟叶和陕西省缴获的烟油中同时检出一种新型合成大麻素:ADB-BUTINACA。我国已经列管了53种合成大麻素~([5]),     

大麻的DNA分析检验技术

本文简要回顾了大麻的一些常规检验方法,并重点综述了大麻植物的DNA分析检验技术,包括随机扩增多态性DNA(RAPD)、测序扩增区段(SCAR)、短串联重复序列(STR)、扩增片段长度多态性(AFLP)、单核苷酸多态性(SNP)等DNA分子标记检测法。并且对这几种技术的应用范围及前景做了简要分析,以期为实践工作提供基础理论帮助。     

RAPD和ISSR分子标记检测大麻的遗传多样性初探

目的利用随机扩增多态性DNA和简单序列重复区间扩增分子标记检测大麻遗传多样性,并探讨其在法医学中的应用价值。方法收集中国4省6个地区的100株大麻叶子样品,采用CTAB法提取基因组DNA,设计选择11个RAPD引物和13个ISSR引物,采用6%中性聚丙烯酰胺凝胶电泳-硝酸银染色法进行检测,根据出现的条带数目和片段大小等分析大麻的多样性。结果 11条RAPD引物扩增出的片段在200bp以上共52条,其中具有多态性的27条;ISSR引物扩增出126条,其中具有多态性的73条;多态性条带比率分别为51.9%和57.9%,其差异不具有统计学意义(P>0.05)。结论 RAPD和ISSR两种方法均可用于大麻遗传多样性分析,对检测毒品原植物的种类和来源地具有一定的应用前景。     

新疆土制大麻烟的社会危害和检验方法

本文对新疆土制大麻烟进行了较系统的定性分析。筛选出土制大麻中性提取的最佳方法和专一的新的显色方法,灵敏度可达0.5—2微克。运用气相色谱、液相色谱可快速准确地分析土制大麻毒品。从而为今后查禁该毒品提供快速、可靠的检测手段。一、麻烟的来源和成份新疆种植的大麻是印度传入的。每年秋季烟毒贩子私自摘取大麻雌株上的花絮、顶尖端的嫩叶、花瓣以及枝托等部位附着的树脂霜为原料晒干后,研成细粉,并集聚成团。即制成大麻毒品——麻烟。     

最高人民法院关于审理毒品案件定罪量刑标准有关问题的解释

为依法严惩毒品犯罪,根据刑法分则第六章第七节的规定,现就审理毒品案件定罪量刑标准有关问题解释如下: 第一条 走私、贩卖、运输、制造、非法持有下列毒品,应当认定为刑法第三百四十七条第二款第(一)项、第三百四十八条规定的“其他毒品数量大”: (-)苯丙胺类毒品(甲基苯丙胺除外)一百克以上; (二)大麻油五千克、大麻脂十千克、大麻叶及大麻烟一百五十千克以上;     

二代测序技术在Y染色体遗传标记分型中的法医学应用

法医遗传学领域常利用Y染色体的父系遗传特点,对非重组区遗传标记进行检测并用于亲缘关系鉴定、混合斑检测、家系排查以及种族推断等研究。目前毛细管电泳仍是应用最为广泛的检测技术,基于该技术的商业化检测试剂盒及数据分析处理系统十分成熟。随着生物信息量的增长,传统检测技术通量低的弊端逐渐显现,推动了法医DNA分型技术的革新。近年来,二代测序(next generation sequencing,NGS)技术发展迅速,其应用已被推广到包括法医遗传学在内的各领域,为Y染色体遗传标记的检测提供了新的技术手段。本文就NGS技术应用于法医学Y染色体遗传标记检测的研究现况和应用前景进行阐述,以期为后续司法实务提供新思路。     

云南大麻DNA的提取及检测初步研究

目的探讨云南大麻DNA的提取及检测方法。方法运用改进的SDS微量法提取大麻总DNA,对所得DNA进行PCR检测。结果用所筛选到的大麻引物检测了云南大麻的DNA遗传标记特征,图谱清晰,结果稳定。结论本方法能有效提取并检测大麻DNA,为实现大麻的品种鉴定、遗传多态性研究及来源追踪提供有价值的科学依据。     

Evaluation of genetic markers for the analysis of THC levels of Cannabis sativa samples using principal component analysis – A preliminary study

Cannabis sativa is a worldwide commercial plant used for medicinal purposes, food and fiber production, and also as a recreational drug. Therefore, the identification and differentiation between legal and illegal C. sativa is of great importance for forensic investigations. In this study, principal component analysis (PCA), an exploratory data analysis technique, was tested to correlate the specific genotype with the concentration of tetrahydrocannabinol (THC) in the samples. C. sativa samples were obtained from legal growers in Piedmont, Italy, and from illegal drug seizures in the Turin region. DNA was extracted, quantified, amplified with a 13-loci multiplex STR and finally analyzed with an automated sequencer. The results showed a trend in the analyzed samples as they differed by their THC content and allele profiles. PCA yielded two clusters of samples that differed by specific allele profiles and THC concentrations. Further validation studies are needed, but this study could provide a new approach to forensic investigation and be a valuable aid to law enforcement in significant marijuana seizures or in tracing illicit drug trafficking routes.     

Whole Plastome Sequences of Two Drug-Type Cannabis: Insights Into the Use of Plastid in Forensic Analyses

DNA is one of the fastest growing tools in forensic sciences, increasing reliability in forensic reports and judgments. The use of DNA has increased in different areas of the forensic sciences, such as investigation of plant species, where plastid DNA has been used to elucidate and generate evidence in cases of traceability of genetically modified and controlled plants. Even with several advances and the practice of using DNA in forensic investigations, there are just few studies related to the identification of genetic tools for the characterization of drug and nondrug-types of Cannabis. Herein, the whole plastomes of two drug-type Cannabis are presented and have their structures compared with other Cannabis plastomes deposited in the GenBank, focusing in the forensic use of plastome sequences. The plastomes of Cannabis sativa “Brazuka” and of the hybrid Cannabis AK Royal Automatic presented general structure that does not differs from the reported for other C. sativa cultivars. A phylogenomic analyses grouped C. sativa “Brazuka” with the nondrug C. sativa cultivars, while the hybrid Cannabis AK Royal Automatic placed isolated, basal to this group. This suggests that the analysis of plastomes is useful toward genetic identification of hybrids in relation to C. sativa.     

人体生物性物质来源鉴定及法医学应用研究进展

犯罪现场中人体生物性物质来源鉴定在重现犯罪过程方面发挥了重要作用,寻找特异性遗传标记鉴定人体不同生物性物质来源是近年来法医学工作者研究的重点和难点。本文就目前研究较多的用于人体生物性物质来源鉴定的遗传标记进行综述,包括DNA甲基化、mRNA、microRNA、微生物菌群、蛋白质等。通过比较不同种类遗传标记鉴定人体生物性物质来源的原理和方法,发现人体不同生物性物质来源的鉴定都有其最适合的遗传标记种类,并且可以采用单一遗传标记或联合多种遗传标记进行检验。尽管目前各法医学实验室无统一的标准和方法鉴定人体生物性物质来源,但研究开发一系列成熟可靠的方法区分人体不同生物性物质,进而发挥其法庭证据作用,将是未来的发展方向。     

线粒体DNA和端粒与年龄相关性的研究进展

用DNA技术从分子水平上推断年龄,已成为法医学和人类学研究领域的新热点。目前用于年龄推断研究的DNA标记主要是线粒体和端粒。本文从线粒体DNA和端粒的概念、线粒体DNA的缺失和突变、端粒长度变化与年龄的相关性等方面对该领域的研究进展进行了综述。旨在为法医实践及进一步的研究提供新的方法和思路。     

Storage of Second World War bone samples: Bone fragments versus bone powder

Bone samples may yield low-quality and low-quantity DNA and duplicated analyses of different genetic markers have to be performed for identification of missing persons. Mostly no DNA extract is left after analyses and efficient storage of bones is needed to ensure the stability of the sample over time for retesting using new markers and new technologies. Usually not all of the bone powder prepared in grinder is used for extraction and rest can be stored for future analyses. After molecular genetic analyses of 88 victims of Second World War (WWII) Konfin I mass grave in Slovenia (performed in 2009), fragments of femurs and bone powder that were left were stored at -20 °C. Some authors reported that long-term storage of powder results in the reduction of DNA preservation and its degradation (even at low temperature), explained by an increase in oxidative damage as a result of the enormous increase in exposed surface area. Consequently, grinding of bones as shortly prior to DNA extraction was recommended. The goal of our study was to explore the difference in DNA yield between bone fragment and bone powder frozen for 10 years. 57 WWII femurs were examined and DNA extracted from each of them using bone fragment (piece sampled next to the one used in 2009) and bone powder obtained in 2009, both taken out of freezer after 10 years of storage. Half gram of bone powder was decalcified using full demineralization extraction method. The DNA was purified in a Biorobot EZ1 (Qiagen) and quantified with PowerQuant kit (Promega). Statistical analysis showed significant difference at the 0.05 level in DNA yield comparing fragments of bones and bone powder stored at -20 °C for 10 years. The results show there is more DNA stored in the bone powder than in the bone fragments. Because of time - consuming powdering procedure we recommend to store not only the fragment of the bone, but obtained bone powder as well.     

Short tandem repeat (STR) DNA markers are hypervariable and informative in Cannabis sativa: implications for forensic investigations

Short tandem repeat (STR) markers are the DNA marker of choice in forensic analysis of human DNA. Here we extend the application of STR markers to Cannabis sativa and demonstrate their potential for forensic investigations. Ninety-three individual cannabis plants, representing drug and fibre accessions of widespread origin were profiled with five STR makers. A total of 79 alleles were detected across the five loci. All but four individuals from a single drug-type accession had a unique multilocus genotype. An analysis of molecular variance (AMOVA) revealed significant genetic variation among accessions, with an average of 25% genetic differentiation. By contrast, only 6% genetic difference was detected between drug and fibre crop accessions and it was not possible to unequivocally assign plants as either drug or fibre type. However, our results suggest that drug strains may typically possess lower genetic diversity than fibre strains, which may ultimately provide a means of genetic delineation. Our findings demonstrate the promise of cannabis STR markers to provide information on: (1) agronomic type, (2) the geographical origin of drug seizures, and (3) evidence of conspiracy in production of clonally propagated drug crops.     

Population data for 94 identity SNPs in four U.S. population groups

The U.S. National Institute of Standards and Technology (NIST) sequenced 1036 human DNA samples from four United States population groups (African American, Asian, Hispanic, and Caucasian) using the ForenSeq DNA Signature Prep Kit with Primer Mix B (DPMB) on a MiSeq FGx instrument. In addition to STR markers, DPMB includes amplification primers for single nucleotide polymorphisms (SNPs) used for individual identification (iiSNPs, n = 94), ancestry inference (aiSNPs, n = 56), and phenotype prediction (piSNPs, n = 22). Resulting sequencing coverage information was interpreted for the 94 iiSNP markers. Here we present performance characteristics of the ForenSeq DNA Signature Prep Kit in the population studied.     

焦磷酸测序技术分析单核苷酸多态性在法医学中的应用

单核苷酸多态性（SNP）是新一代的法医学遗传标记,有望成为法医实践中解决高度降解检材及特殊案件DNA鉴定的重要工具。近年来涌现出许多高通量的SNP分析方法,如引物延伸结合时间飞行质谱分析法、微测序法、SNP lex以及焦磷酸测序法等。本文重点对焦磷酸测序技术的原理、步骤及其在法医学中的应用进展进行简要的综述。