DNA指纹图在法医学鉴定中应用的研究(Ⅱ)——应用‘Myo’小卫星探针进行血斑、精斑、个体组织的个体认定

应用‘Myo’小卫星 DNA 探针,Southern 印迹杂交技术,对血斑、精斑、同一个体不同组织进行 DNA 指纹图分析,均获得清晰的图谱。同一个体的血斑与血液、精斑与精液以及不同的组织其 DNA 指纹图谱完全相同。可以根据斑痕或组织与嫌疑个体的血液或某一组织 DNA 的指纹图谱比对以做出同一认定。50μl 血液量的血斑、5μl 精液量的精斑可以获得清晰易辨的指纹图谱。五年的精斑、两年的血斑亦可做出与同源个体新鲜精液、血液完全一致的 DNA 指纹图谱。对杀人、强奸杀人、碎尸等不同案件的血痕、精斑、不同组织碎块进行了 DNA 指纹图检验,均做出了正确的个体认定。本方法的应用为我国法医物证检验提供了新的分析手段,使个体认定得以实现。     

用引物Y_3,Y_4和DNA扩增技术作血液(痕)和毛根性别鉴定的研究

作者用引物Y_3、Y_4和DNA聚合酶链式反应(PCR)作微量人类血液(痕)和毛根的性别鉴定。扩增的靶序列位于Y染色体DNA特异3.4kb重复序列中,扩增产物为460bp。检材用量为:新鲜血液0.5μl、血痕纱纤维1mm、毛根单个。20例保存4个月的血痕与2例保存6年半的血痕性别判定结果均正确,无性别记载的保存9～11年的3例血痕显现了清晰的460bpY特异DNA扩增带。15例保存20天的自然脱落毛根性别判定结果均正确。本法省略了检材处理中的酚-氯仿抽提DNA等纯化步骤,既简化了实验操作,又减少了检验过程中外源DNA的污染机会和样品DNA的损耗,使这一性别鉴定方法更符合法医学实践的需要。     

Generating DNA profiles from immunochromatographic cards using LCN methodology

The aim of this research was to obtain DNA profiles from immunochromatographic test devices which have already yielded positive results with body fluids obtained from fourteen volunteers. Three different immunochromatographic cards for the identification of human blood and one for the identification of human saliva were used for this research. Each body fluid was detected using the appropriate immunochromatographic card. The used cards were kept at room temperature for various lengths of time. The membranes were removed at the end of the designated times and the entire strip was extracted using low copy number (LCN) extraction procedure. The extracted DNA was amplified using reduced amplification volume and higher PCR cycle numbers. Autosomal STR profiles were detected using AmpFℓSTR^® Identifiler™ PCR Amplification Kit from Applied Biosystems (AB). Additionally, DNA extracted from the male volunteers was amplified using the AB AmpFℓSTR^® Yfiler™ PCR Amplification Kit. Analysis of the amplified products was carried out by capillary electrophoresis injection on the AB 3130xl Genetic Analyzer. The generated DNA data was analyzed using the SoftGenetics GeneMarker^® HID Version 1.7 software.Autosomal and Y-STR DNA profiles were obtained from most of the cards which were stored at room temperature for up to three months. DNA profile was obtained from all four types of the immunochromatographic cards used in this study. These profiles were concordant with the profiles obtained from the donors’ reference samples.     

Human identification and sex determination of dental pulp, bone marrow and blood stains with a recombinant DNA probe

Recombinant DNA hybridizing specifically to a 300 nucleotide repeat DNA sequence (BLUR8) of human specificity and to human repeat DNA sequence (pHY10) on the Y chromosome was used for human identification and sex determination of degraded DNA samples of blood stains, dental pulp, and bone marrow. This radioactive technique enabled reliable and sensitive human and sex determination from blood stains that were more than 80 years old. Less than 1 piece of 0.5 cm length thread of blood stain was enough for both tests. DNA from relatively fresh dental pulp and bone marrow was clearly identified. The human identification test, which could recognize up to 0.3 ng DNA correctly, was 3 to 5 times more sensitive than the sex determination test.     

Classification of prepaid cards based on multivariate treatment of data obtained by X-ray fluorescence analysis

Two hundred prepaid cards, which had been used in Nagoya-city's subway in Japan, and another 32 prepaid cards (11 were real turnpike cards, 20 were counterfeit cards and 1 was a white card) were evaluated by X-ray fluorescence (XRF) without any pre-treatment. A preliminary investigation was performed on 200 prepaid cards in order to find an identification method for the turnpike cards. By plotting the relative intensity of titanium versus that of iron obtained by XRF, the cards were clearly classified into seven groups. On the other hand, the cards could be divided into four groups by a multivariate analysis using the relative intensities of five elements such as chlorine, calcium or tin, titanium, iron and barium. Using these results to classify the Japanese turnpike cards, they were divided into three groups or two groups. One of three groups or two groups was the counterfeit card group.     

信用卡诈骗罪若干问题研究

信用卡诈骗罪是以信用卡作为犯罪工具的诈骗犯罪。建议该罪亦可由单位构成犯罪主体 ;骗领信用卡人不能成为恶意透支的犯罪主体 ;对使用伪造、作废信用卡、冒用他人信用卡 ,恶意透支行为 ,应区别不同情况分别处理 ;危害信用卡并使用的行为 ,拾得信用卡并使用的行为构成本罪 ;对骗领信用卡并使用的行为、信用卡协议透支的行为 ,应区别不同情形 ,作出相应处罚。     

应用抗人精液单克隆抗体检验混合斑中精液成份的ABH血型物质

用本文报道的方法检测了123份混合斑检材（新鲜混合斑60份，陈旧混合斑63份），对保存在一年内的混合斑中精液成份中的血型物质检出率可达100％。     

Internal validation of reduced PCR reaction volume of the Qiagen Investigator® Argus X-12 QS Kit from blood samples on FTA cards

The onus of proof in criminal cases is beyond any reasonable doubt, and the issue on the lack of complete internal validation data can be manipulated when it comes to justifying the validity and reliability of the X-chromosomal short tandem repeats analysis for court representation. Therefore, this research evaluated the efficiency of the optimized 60% reduced volumes for polymerase chain reaction (PCR) amplification using the Qiagen Investigator® Argus X-12 QS Kit, as well as the capillary electrophoresis (CE) sample preparation for blood samples on Flinder's Technology Associates (FTA) cards. Good-quality DNA profile (3000–12,000 RFU) from the purified blood sample on FTA card (1.2 mm) were obtained using the optimized PCR (10.0 μL of PCR reaction volume and 21 cycles) and CE (9.0 μL Hi-Di™ Formamide and 0.3 μL DNA Size Standard 550 [BTO] and 27 s injection time) conditions. The analytical and stochastic thresholds were 100 and 200 RFU, respectively. Hence, the internal validation data supported the use of the optimized 60% reduced PCR amplification reaction volume of the Qiagen Investigator® Argus X-12 QS Kit as well as the CE sample preparation for producing reliable DNA profiles that comply with the quality assurance standards for forensic DNA testing laboratories, while optimizing the analytical cost.     

Validation studies of rapid stain identification-blood (RSID-blood) kit in forensic caseworks

When bloodstains are detected at crime scene using presumptive tests (e.g. luminol, phenolphthalein, leuchomalachite green), it is important to establish the real human nature of each stain. This is possible using confirmatory tests. One of these is rapid stain identification-blood (RISD-blood) a lateral flow immuno-chromatographic strip test format which allows the identification of human blood by detection of glycophorin A, a red blood cell membrane antigen, using two anti-human glycophorin A (GPA) monoclonal antibodies.The aim of this study is to assess the sensitivity of RSID-blood test in old, degraded bloodstains and in some bloodstains previously treated with BlueStar Forensic, a presumptive test which is often used in crime scene investigations to detect latent bloodstains. The genetic analysis of all bloodstains of confirmed human nature was subsequently performed using the AmpF1STR Identifiler PCR Amplification Kit (Applied Biosystems), to validate the possibility of obtain a consistent and reliable DNA typing results.     

PCR扩增ZFY/ZFX基因鉴定人类干血痕性别

应用 PCR 技术同时扩增人 ZFY 和 ZFX 基因特异的 DNA 序列,在男性血痕中可检测到两种扩增产物,即340bp 长的 ZFY 基因及488bp 长的 ZFX 基因特异 DNA 片段;在女性血痕中仅可检测到488bp 长的 ZFX 基因特异 DNA 片段,据此判定干血痕性别。干血痕的最小检出需要量为0.125μl 血液量的血痕。室温保存10年的血痕可以准确判定性别。ZFY 基因位于 Y 染色体短臂。本方法同时检测两条性染色体,可以避免由于扩增失败或 Y 染色体长臂变异出现的假阴性或假阳性。扩增产物经琼脂糖凝胶电泳即可区分。     

Cranial Backspatter Pattern Production Utilizing Human Cadavers

A backspatter pattern results from blood drops that travel retrograde to an applied external force. Historically, an array of animals and nonhuman objects have been used to create and study backspatter patterns. In this study, backspatter patterns captured on foam core targets that were placed 45.72 cm (18 in) behind the impact site (occipital area of the skull) were produced by cranial gunshots to human cadavers that were reinfused with fresh defibrinated bovine blood. These patterns were compared to the backspatter patterns produced by shooting blood‐soaked sponges, a typical simulant used in controlled studies of backspatter pattern production and characteristics. The backspatter pattern produced by shooting an actual human head was found to be different than those of blood‐soaked sponges in the number of stains produced, the size and size range of the stains, and the stain dispersion patterns.     

Forensic data recovery and examination of magnetic swipe card cloning devices

Magnetic swipe card technology is used for many purposes including credit, debit, store loyalty, mobile phone top-up and security identification cards. These types of cards and the details contained on them are often relied upon as a form of identification and personal authentication. As such reliance is placed upon them it is surprising that they do not incorporate more stringent security features, and because of this lack of features it is not surprising that they attract the attention of people who wish to exploit them for illegal gain. The paper introduces the type of technology, and range of devices available for manipulating magnetic swipe card data. It proposes the use of Digital Evidence Bags as a suitable format for the evidential storage of information obtained from them, thus further illustrating the flexibility of the format and demonstrating the diverse range of devices that have to be handled within the digital investigation and law enforcement community.     

Field testing of collection cards for Cannabis sativa samples with a single hexanucleotide DNA marker

The validity and feasibility of using DNA collection cards in the field for preservation and analysis of Cannabis sativa genotypes were investigated using a highly specific hexanucleotide marker. Collection cards were submitted to the National Marijuana Initiative, which selectively trained and managed the collection of specific types of samples from a variety of participating agencies. Samples collected at seizure sites included fresh marijuana leaf samples, dried "dispensary" samples, U.S. border seizures, and hashish. Using a standardized PCR kit with custom-labeled oligonucleotide primers specific to marijuana, collection cards produced eight genotypes and 13 different alleles, extremely low baselines, and no cross-reactivity with control plant species. Results were produced from all sample types with the exception of hashish. Plant DNA collection cards represent an easily implementable method for the genetic identification and relatedness of C. sativa street and grow site-seized samples with applications for databasing and market disruption.     

伪造信用卡犯罪中的伪造行为内涵与对象研究

刑法中伪造信用卡犯罪中的伪造不仅包括仿制其物理外观的形式伪造,还应包括将权利人信息写入磁条介质、芯片等的内容伪造。作为伪造对象的信用卡应当是具有物理载体的实体卡片,事实上不存在对虚拟信用卡的伪造。以虚假身份骗领无对应实体卡的虚拟信用卡应视为妨害信用卡管理罪中的骗领信用卡行为;利用权利人既有实体卡信息,复制虚拟信用卡的行为系对他人信用卡信息资料的非法使用而非伪造。有关伪造信用卡犯罪刑法规定的立法原意并不包含伪造空白信用卡,伪造空白信用卡具有严重的社会危害性,理应将其纳入刑法规制范围,以保持与妨害信用卡管理罪刑法规定的协调。不能通过司法解释改变立法原意,现行刑法中伪造信用卡犯罪的伪造含义应有相应的调整。     

FTA卡在微量血痕DNA多态性分析中的应用

目的探索FTA卡样本DNA最小检出量及同一FrA卡样本进行多项目检测的可行性。方法制作含有不同浓度DNA丌A卡,使用Identifiler试剂盒进行检验;对同一rrA卡样本先后使用Identifiler、Y-filer试剂盒进行扩增及线粒体高变区HVI区测序,使用ABl3130测序仪检测各扩增产物。结果载有0．5ngDNA的FTA卡扩增产物即可正确分型,对同一血痕样本使用不同试剂盒检验,均可清晰正确判型,线粒体高变区HVI区测序图清晰,且各检验结果均与对照一致。结论载有0．5ngDNA的FTA卡扩增产物可用于DNA分型检测,FTA卡对DNA结合较稳定,可用于多项目检测。     

Successful DNA typing of a urine sample in a doping control case using human mitochondrial DNA analysis

In a doping control case, a urine sample was tested positive for nandrolon. We were asked by the athlete to perform DNA investigations on the questioned urine sample and compare these to a fresh blood sample taken from the athlete in order to detect or rule out manipulation and/or switching of the samples. The urine sample had been collected nine months prior to the investigation and had been stored at 4 degrees C. In a first approach, nuclear DNA systems were investigated that failed with the exception of the Amelogenin system. Due to the high copy number of mitochondrial DNA molecules and the robustness of the mitochondrial genome, we investigated the HVR I and HVR II regions of mitochondrial DNA and obtained reproducible and clear sequencing results for both the blood and the urine samples. Due to the identical sequences, it could not be excluded that the blood sample and the urine sample were from the same individual or an individual having the same maternal lineage.     

Identification of a skeleton using DNA from teeth and a PAP smear.

Identification of unknown living or deceased persons using dental treatment records is an established forensic technique. However, some cases remain unidentified, especially when antemortem dental records are not available for comparison to postmortem dental records. Cytological smears have been previously reported to be potential sources of DNA reference samples which can be compared to DNA recovered from found human remains. The case described here involves an adult skeleton which exhibited extensive, complex dental restorative treatment. A putative identification of the found skeleton as a missing woman was established using circumstantial evidence found at the scene. However, it became important to establish a positive identification using reliable scientific methods. When it was discovered that antemortem dental records were not available because the treatment was completed in another country and the treating dentist could not be found, cytological smears stained with Papanicolaou (PAP) stain obtained from the putative decedent's medical records were used as a reference DNA sample. DNA was recovered from the teeth of the skeleton using cryogenic grinding. Comparison of the genotypes resulted in the conclusion that the DNA originated from the same source. The use of PAP smears in this way is seen as a valuable resource in cases where positive identification using traditional dental and medical records is not possible.     

Evaluating the use of hypoxia sensitive markers for body fluid stain age prediction

To augment DNA profiling and body fluid identification techniques efforts are being made to increase the amount of information available from a crime scene stain, which includes efforts to identify externally visible characteristics through phenotypic analysis. A key question surrounding crime scene stains is the length of time between deposition of the stain and its subsequent recovery, in that is the stain recovered related to the incident in question or from a previously deposited stain number of weeks earlier? The inability to answer this fundamental question has a detrimental effect upon the successful completion of a criminal investigation. Once a body fluid leaves the body, the oxygen concentration in the environment changes; therefore, it may be that this change could cause a change in the expression of hypoxia-sensitive biomarkers. Here, a range of bloodstains, liquid saliva and liquid semen samples were collected at 0 days, 7 days, 14 days, 21 days and 28 days of degrading at room temperature (19–22 °C), before undergoing total RNA extraction and cDNA synthesis. Blood was recovered from filter paper with 3 mm², with saliva and semen being left in their tubes and swabbed at the appropriate times. All samples then underwent quantitative PCR targeting Vascular Endothelial Growth Factor A (VEGFA) and Hypoxia-Inducible Factor 1 Alpha (HIF1A), with B-Actin (ACTB) as a reference gene. A range of linear and quadratic correlation values was obtained from the qPCR data and used to develop a predictive model with a mean absolute deviation (MAD) of 4.2, 2.1, and 5 days for blood, saliva, and semen respectively. Blind testing indicated that a stain age prediction model based upon VEGFA with ACTB as a reference gene could be used on samples up to four weeks old with a margin of error ranging from 2 days through to 5 days. While a sizeable potential time frame exists using this model; this represents a significant step towards the target of having an accurate stain age prediction model.     

SIM卡中信息提取方法的研究

目的建立从SIM卡中提取具有证据价值的数据,包括删除数据的原理及方法。方法通过Paraben Cell Seizure软件(版本2.0)及其配套的读卡器,读取SIM卡中现存和删除的数据信息。结果从实验用的SIM卡中可恢复出电话记录和短信等一系列数据。结论SIM卡中包含大量有价值的信息,这些信息中的多数都可以再现和利用,有作为证据的价值。