JL-02多位点探针DNA指纹的法医学应用研究

以自制的JL－02探针进行了DNA指纹分析，对北京地区无关个体进行了调查，计算出任意两无关个体的偶合机率为6．6×10－15；家系分析表明，谱带在亲代与子代间的传递符合孟德尔遗传规律；同一个体不同组织的DNA指纹图相同；混合斑精子DNA指纹图与相应男性血液DNA指纹图完全相同；该探针对0．5μg的基因组DNA杂交，可获得清晰可辩的DNA指纹图。证明了新探针适用于法医物证检验中的个人同一认定及亲子鉴定。     

33.15及33.6探针对中国人DNA指纹图的研究

目的统计用33．15及33．6探针对中国人进行DNA指纹图检验的群体调查资料，为实际案件的检验提供理论依据。方法应用33．15及33．6探针为中国北京地区无关群体的血液进行DNA指纹图分析。结果应用33．15探针检验15人的DNA指纹图，无关个体间相关机率为1．03×10-15，两无关个体间出现同一谱带的平均概率0．176；应用33．6探针检验19人的DNA指纹图，无关个体间相关机率为1．53×11-11，两无关个体间出现同一港带的平均概率为0．187；两探针均符合孟德尔遗传规律，均具有组织同一性。结论本研究结果可应用于法医物证检验的亲子鉴定及个体识别。     

用α-珠旦白-3’HVR探针研究DNA指纹

用α-珠蛋白-3’HVR探针,经Southern印迹法,对100名不相关个体及4个家系的32名相关个体的DNA指纹进行了检测,所产生的DNA图谱具有高度的个体特异性,在被测的所有个体中无一相同。经统计学计算表明,任意两个个体DNA指纹图重合率为10~(-11)。家系分析毒明,DNA片段严格按照孟德尔方式遗传。该探针的应用,将在法医学亲子鉴定和个人同一认定中发挥重要的作用。     

DNA技术在法医学上的应用(续)

<正> 四、DNA指纹用于个体识别因DNA具有高度多态性,因此不同个体DNA指纹图各不相同,可以进行个体识别。下面以Jeffreys的33.15DNA探针做DNA指纹图为例,说明DNA指纹图的个体识别作用。取20名随机个体DNA,以HinfI消化,Southern印迹后与33.15探针杂交,得每个人DNA指纹图。然后做相邻两个体指纹图比较,将20个个体图中区带数全部算出,取平均值、并计算标准差。然后根据相邻个体共存区带数,求出等位基     

微型DNA指纹图的研究与应用

作者采用小板琼脂糖凝胶电泳,应用MYO探针,Southern印迹杂交技术制作微型DNA指纹图,能获得清晰图谱。实验结果表明,同一个体的血液与血斑,精液与精斑及不同部位的组织,其指纹图谱完全相同。不同个体的微型DNA指纹图谱有明显个体差异。用本法能使大板DNA指纹图的常规检出量重复15～20次,最小检出量为250ng,达到极微量的水平。     

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。     

DNA指纹技术在强奸案和亲子鉴定中的应用

在同一张膜上测定了18例无关个体血样,并计算出α-珠蛋白-3'HVR探针DNA指纹图的相关机率为4.0×10~(-12),平均每条谱带的相关机率为0.21。用此探针检验的14起强奸案和6起亲子鉴定案均获得肯定的结论。     

DNA纹印技术的初步研究——用PYNH24探针分析MspI的RFLP

应用 Southern 印迹杂交技术,分析了人体基因组 DNA MspI 酶解物与特异探针 PYNH24杂交的限制酶切片段长度多态性,获得清晰易读的杂交图谱。50个不相关个体血样中未发现相同图谱,显示了个体特异性,为法医生物物证检验工作提供了新的分析手段。     

非同位素标记DNA探针初步研究

本文报导了将辣根过氧化物酶直接标记在单链 DNA 探针上,制备出非同位素的 DNA 探针,采用化学光增强法检测杂交结果,所得图谱清晰,容易判读。所制备出的 DNA 探针可用于人类性别鉴定和个体识别,为 DNA 检验技术推广应用开辟了新的领域。     

33.15及33.6探针对中国人DNA指纹图的研究

目的 统计用３３．１５及３３．６探针对中国人进行ＤＮＡ指纹图检验的群体调查资料，为实际案件的检验提供理论依据。方法 应用３３．１５及３３．６探针为中国北京地区无关群体的血液进行ＤＮＡ指纹图分析。结果 应用３３．１５探针检验１５人的ＤＮＡ指纹图，无关个体间相关机率为１．０３&#215;１０＾－１５，两无关个体间出现同一谱带的平均概率０．１７６；应用３３．６探针检验１９人的ＤＮＡ指纹图，无关个体间相关机率为１．     

DNA指纹图在法医学鉴定中应用的研究(Ⅲ)——应用“Myo”DNA探针进行混合斑中精液的个体认定

本文利用蛋白酶K、SDS对精液和阴道液、精液和血液的混合斑进行前处理,除去女性阴道脱落上皮细胞和血液细胞成份获得精子。提取精子DNA,用“Myo”小卫星DNA探针杂交进行DNA指纹检验,获得了高度多态性的精子DNA指纹图谱,与同一个体血液DNA指纹图谱比较完全一致,实现了混合斑中精液来源的个体认定。在对20多起强奸案例混合斑的实际应用中,成功地认定了强奸罪犯。     

人DNA指纹检测的初步研究

根据随机探针检测DNA限制片段长度多态性的原理和人与鼠的髓鞘硷性蛋白(MBP)基因cDNA有90％以上同源序列的事实,我们选用rMBP-cDNA 3'端非表达区高度重复顺序的0.81kb片段作探针,检测用HaeⅢ酶解的人DNA限制性片段结果可以分解出22条谱带,受检的30例无血缘关系的个体之间,没有两个人的谱带是完全相同的,显示出此方法的高度特异性。本文还比较了若干DNA片段作探针和几种限制性内切酶检测人DNA指纹的结果。     

A novel method for ABO genotyping using a DNA chip

ABO genotyping is often performed to identify the blood type of decomposed samples, which is difficult to be determined by a serological test. In this study, we developed a simple method for ABO genotyping using a DNA chip. In this method, polymerase chain reaction-amplified and fluorescent-labeled fragments in the ABO gene and primate-specific D17Z1 were hybridized with DNA probes on a chip designed to detect single nucleotide polymorphisms (SNPs) in the ABO gene and part of the D17Z1 sequence. Using blood samples from 42 volunteers and 10 animal species, we investigated whether the chip could be used to detect SNPs in the ABO gene and the D17Z1 sequence. This method was then applied to various forensic samples, and it was confirmed that this method was suitable for the simultaneous analyses of ABO genotyping and species identification. This method fulfills the recent need for the development of rapid and convenient methods for criminal investigations.     

New optimized DNA extraction protocol for fingerprints deposited on a special self-adhesive security seal and other latent samples used for human identification

Abstract: Obtaining complete short tandem repeat (STR) profiles from fingerprints containing minimal amounts of DNA, using standard extraction techniques, can be difficult. The aim of this study was to evaluate a new kit, Fingerprint DNA Finder (FDF Kit), recently launched for the extraction of DNA and STR profiling from fingerprints placed on a special device known as Self‐Adhesive Security Seal Sticker^® and other latent fingerprints on forensic evidentiary material like metallic guns. The DNA extraction system is based on a reversal of the silica principle, and all the potential inhibiting substances are retained on the surface of a special adsorbent, while nucleic acids are not bound and remain in solution dramatically improving DNA recovery. DNA yield was quite variable among the samples tested, rendering in most of the cases (>90%) complete STR profiles, free of PCR inhibitors, and devoid of artifacts. Even samples with DNA amount below 100 pg could be successfully analyzed.     

Highly polymorphic minisatellite DNA probes. Further evaluation for individual identification and paternity testing

Reliable and reproducible protocols have been developed for the routine DNA fingerprinting of individuals using the highly polymorphic minisatellite DNA probes 33.15 and 33.6. Comparison of DNA fingerprinting from 50 individuals has generated further data on the level of band sharing in the DNA fingerprints of unrelated individuals, as well as the number of bands scorable in individuals. These results are consistent with previous studies. The occurrence of mutant bands in offspring has been examined in over 100 families. Further support is presented for the Mendelian inheritance of minisatellite loci and for lack of significant allelism and linkage between different variable DNA fragments detected in a human DNA fingerprint.     

Establishing an integrated pipeline for automatic and efficient detection of trace DNA encountered in forensic applications

The analysis of trace DNA is a crucial component in forensic applications. Biological materials containing low-level DNA collected at crime scenes, such as fingerprints, can be valuable as evidence. Automatic detection of biological samples has been largely embraced in forensic applications to meet the increasing throughput requirements. However, the amount of DNA automatically retrieved from trace evidence often tends to be small and unstable, ultimately resulting in poor detection of DNA profiles. Thus, in this work, we introduced a robust DNA extraction and purification platform named Bionewtech® BN3200 (Bionewtech®, Shanghai, China) with the goal of constructing a rapid automatic detection system for trace DNA. The establishment of automatic detection system for trace DNA mainly encompassed two parts: assessing the sensitivity of automatic extraction platform and screening the optimal short tandem repeat (STR) typing kit. The sensitivity of Bionewtech® BN3200 platform based on Ultra-sensitive DNA Extraction kit was initially estimated, demonstrating that this extraction platform might contain large potential in the trace DNA extraction. For the amplification part, three sets of commercial multiplex STR typing kits were selected as candidates, and the amplified products were further genotyped on the Applied Biosystems 3500xl Genetic Analyzer. After comparation, SiFa™ 23 Plex Kit was determined as the most suitable amplification system for trace DNA. Eventually, the newly exploited trace DNA detection system was successfully implemented in the detection of fingerprints derived from glass surfaces with the five-seconds contact time. As a result, the DNA recovered from the fingerprints fluctuated approximately from 57.60 pg to 18.05 ng, in addition, over 70% of the total STR loci were detected in 75% of the fingerprint samples.     

探讨EOS染色剂对血迹DNA检验的影响

目的探讨被EOS染色剂处理后的血迹进行DNA检验的初步方法。方法制备EOS染色剂处理的血迹样本，分别采取纯水擦拭、75％酒精擦拭，或手术刀刮取血痕浸泡于纯水中、759／5酒精中，然后提取DNA进行下一步检测。结果采用刀刮取血痕浸泡于759／6酒精中，提取的DNA检测结果较好。结论初步实验显示，经EOS染色剂处理过的血迹，可参考刀刮取血迹置于75％酒精浸泡后提取DNA做下一步检测。     

Analysis of restriction fragment length polymorphisms in deoxyribonucleic acid (DNA) recovered from dried bloodstains

Deoxyribonucleic acid (DNA) was recovered from dried bloodstains aged up to three years and shown to be of high molecular weight. DNA was digested with restriction endonucleases and fractionated by agarose gel electrophoresis. Following transfer to a filter, DNA was hybridized with two different radioactively labeled recombinant probes which recognize highly polymorphic regions in human DNA. The autoradiographic pattern observed was not altered by sample age, and the size of the alleles was consistent with those observed in the general population. Therefore, DNA of high molecular weight prepared from dried blood samples can be used for identification.     

An Optimized DNA Analysis Workflow for the Sampling,Extraction, and Concentration of DNA obtained from Archived Latent Fingerprints

DNA profiles have been obtained from fingerprints, but there is limited knowledge regarding DNA analysis from archived latent fingerprints—touch DNA “sandwiched” between adhesive and paper. Thus, this study sought to comparatively analyze a variety of collection and analytical methods in an effort to seek an optimized workflow for this specific sample type. Untreated and treated archived latent fingerprints were utilized to compare different biological sampling techniques, swab diluents, DNA extraction systems, DNA concentration practices, and post‐amplification purification methods. Archived latent fingerprints disassembled and sampled via direct cutting, followed by DNA extracted using the QIAamp® DNA Investigator Kit, and concentration with Centri‐Sep? columns increased the odds of obtaining an STR profile. Using the recommended DNA workflow, 9 of the 10 samples provided STR profiles, which included 7–100% of the expected STR alleles and two full profiles. Thus, with carefully selected procedures, archived latent fingerprints can be a viable DNA source for criminal investigations including cold/postconviction cases.