复合扩增D16S539、D7S820、D13S317基因座的DNA分型及法医学应用

通过对3个位于不同染色体上的STR基因座（D165539,D7S820,D13S317）所组成的复合扩增体系的DNA分型研究,以期在实际法医物证检验中增加检验基因座,以提高总的个体识别率。笔者运用复合扩增技术,经4％变性聚丙烯酸胺凝胶电泳分离扩增产物和银染检测,首次对108个无关中国人个体的D16S539,D7S820,D13S317基因座进行研究,检测出中国人群中3个基因座的等位基因数均为7个;偶合率P（m）分别为0．0847、0．0740、0．0741;个体识别率DP值分别为0．9153、0．9260、0．9259;杂合度分别为77．7％、79.1％、79．3％;各基因座亲子关系指数PItypical分别为2．24、2．39、2．42。3个STR基因座总的个体识别率很高,达0．9995;总的亲子关系指数PItypical达12．96;所有基因座经卡方检验符合Hardy－Weinberg平衡。通过以上数据可以看出,D165539,D7S820,D13S317基因座所组成的复合扩增体系在中国人群中等位基因分布较好,个体识别率很高,适合用于法医个体识别及亲子鉴定。     

Genetic analysis of 15 STR loci in the population of Zhejiang Province (Southeast China)

Fifteen autosomal STR loci were analyzed from a population sample of 598 unrelated individuals residing in Zhejiang Province. We report allele frequencies distribution and statistical parameters for all 15 STR loci, D8S1179, D21S11, D7S820, CSF1PO, D3S1358, THO1, D13S317, D16S539, D2S1338, D19S433, VWA, TPOX, D18S51, D5S818 and FGA. Allele frequencies, the observed heterozygosity (Ho), the polymorphic information content (PIC), and the probability of paternity exclusion (PE) were calculated. All loci were in accordance with Hardy–Weinberg equilibrium (P > 0.05). Our studied population data were compared with the previously published population data of other ethnic groups or areas in China. Our results of present study were valuable for human identification and paternity tests in Zhejiang Province.     

The Upper Silesia (Poland) population and forensic usefulness of 15 autosomal STR loci

Allele frequencies, forensic parameters for the 15 STR loci in the AmpFlSTR^® Identifiler Kit (Applied Biosystems), D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D19S433, VWA, TPOX, D18S51, D5S818 and FGA were determined in a sample of 150 unrelated dead and alive adults from the Upper Silesia region (Poland). The values of heterozygosity (Ht), polymorphic information content (PIC), power of discrimination (PD), matching probability (PM), mean exclusion chance (MEC) and mean exclusion probability (MEP) were calculated. Possible divergence from HWE was determined. Comparison of allele frequencies for examined STR loci between the Upper Silesia population and other Polish populations was carried out.     

Human Y-specific STR haplotypes in a Slovenian population sample

The allele distribution of the systems DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS385 and YCAII were investigated in a sample of 121 unrelated males from Slovenia     

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.     

Interrogation of short tandem repeats using fluorescent probes and melting curve analysis: A step towards rapid DNA identity screening

Current forensic DNA profiling methods rely on the analysis of samples at specialised laboratories with an average turnaround time of several days. The ability to rapidly determine a partial profile of short tandem repeats at the point-of-arrest would be of great benefit to police forces around the world, for example enabling a suspect to be rapidly included or excluded from an investigation. We have developed a homogeneous PCR method for the interrogation of STR loci utilising fluorescent oligonucleotide probes and melting curve analysis. Alleles of the D18S51, TH01 and D8S1179 loci were differentiated and identified on the basis of target length and probe melting temperature. Assay performance was evaluated by comparing melting peak data with the AmpFlSTR^® SGM Plus^® system. The method is compatible with direct analysis of unpurified buccal swab samples, enabling a partial STR profile to be generated within 1 h.     

复合扩增STR位点片段长度多态性的研究

本文介绍了以三个短串联重复序列（STR）位点为基础的复合扩增进行个体识别的技术。其特点是在同一扩增体系内对彼此独立的三个STR位点，即HUMTH01[AATG]nHUMFABP[AAT]nHUMARA[AGC]n进行复会扩增，扩增产物用聚丙烯酰胺凝胶电泳分离。经研究发现这三个STR位点具有高度多态性，扩增产物分子量在190～310bP之间，各位点等位基因数分别为6、9、16个，杂合度78．2％、85，0％、89．0％，个体识别率0．918、0．960、0．905，其累积的个体识别率达0．9997，高于pMCT118等位点，且扩增时间短，操作简便，为法医物证检验提供了一种高效的个体识别手段。     

New resources for the forensic genetics community available on the NIST STRBase website

For more than a decade, the U.S. National Institute of Standards and Technology (NIST) has maintained the short tandem repeat DNA Internet database (STRBase), which is located at http://www.cstl.nist.gov/biotech/strbase/. The purpose of STRBase has been and continues to be an attempt to bring together the abundant literature and information in the forensic genetics field in a cohesive fashion to make current and future work easier. New materials are regularly added to expand the valuable information contained on the STRBase website.     

Autosomal and Y-STR mutations in Lithuanian population

The new approach is given in this work combines autosomal and Y-chromosomal short tandem repeats (STRs) data in paternity testing with father/son pairs taking into account mutation events. Our results show that Y-STRs can identify nonpaternity cases with high accuracy but need to be validated with autosomal STR to establish paternity. Autosomal and Y-STR mutations give disputing questions in paternity testing as well.     

STR data for the AmpFℓSTR Identifiler loci from Swedish population in comparison to European, as well as with non-European population

The modern Swedish population is a mixture of people that originate from different parts of the world. This is also the truth for the clients participating in the paternity cases investigated at the department. Calculations based on a Swedish frequency database only, could give us overestimated figures of probability and power of exclusion in cases including clients with a genetic background other than Swedish. Here, we describe allele frequencies regarding the markers in the Identifiler-kit. We have compared three sets of population samples; Swedish, European and non-European to investigate how these three groups of population samples differ. Also, all three population sets were compared to data reported from other European and non-European populations.Swedish allele frequencies for the 15 autosomal STRs included in the Identifiler kit were obtained from unrelated blood donors with Swedish names. The European and non-European frequencies were based on DNA-profiles of alleged fathers from our paternity cases in 2005 and 2006.     

Case study of dissolving allele dropout

Allele typing performed using the original components of the AmpFlSTR Identifiler kit led to unbalanced peak heights and allele dropouts. However, allele typing was completely successful when substituting the QIAGEN Multiplex PCR Master Mix for the AmpFlSTR PCR Reaction Mix of the Identifiler kit. The successful range for allele typing was 250 pg to 1.5 ng of DNA. The running times for PCR amplification were reduced by about 30 min from the original protocol.     

European validation of a Cannabis sativa 13-locus STR multiplex kit for genetic identification: A preliminary study

Cannabis sativa L. is a plant cultivated worldwide as a source of fiber, medicine and intoxicant. Traditionally, is divided into two main types: fiber type (hemp) and drug type (marijuana). Marijuana differs from hemp by the presence of a high quantity of the psychoactive drug, Δ9-tetrahydrocannabinol. The development of a validated method using short tandem repeats (STRs) could serve as an intelligence tool to link cases by means of genetic individualization or association of cannabis samples. For this purpose, a 13-locus STR multiplex method was developed, optimized, and validated by the Department of Forensic Science at Sam Houston State University (SHSU) according to relevant ISFG and SWGDAM guidelines. The European community considers C. sativa plants illegals, even though its consumption is accepted in precise and limited places (coffee shops or cannabis clubs in Netherlands and Spain). However, there are different gaps in the legislation of some European countries. For instance, in Italy, “weed” possession is decriminalized. Although trafficking and sale are prohibited, possession of small quantities of marijuana is considered only a civil offense. In order to proceed with the kit evaluation and inter-laboratory comparison, SHSU DNA laboratory sent blind cannabis DNA samples of known genotypes. Blind DNA samples were analyzed in different laboratories with different sequencers and analysis conditions. In this article, the goals were: a) to demonstrate that 13-locus STR kit for C. sativa is robust enough and reproducible, in all forensic laboratories, and b) to show the applicability of the STR system in association with Cannabis sativa cases for intelligence purposes to link multiple cases by means of genetic individualization or association of cannabis samples.     

Fast PCR amplification of AmpFlSTR Identifiler

To increase throughput for DNA typing, we examined fast PCR cycling using AmpFlSTR Identifiler by three methods. In this study, we reduced PCR running times by 1/3 to 2/3 (approximately 1-2 h). This means DNA typing, including PCR reactions, can be completed within a timeframe ranging from 90 min to 2 h and 30 min.     

河南汉族群体6个STR基因座遗传多态性研究

目的　通过研究 6个STR基因座FGA ,TPOX ,D3S135 8,vWA ,D8S1179,D2 1S11的遗传多态性 ,了解它们在河南汉族人群中的多态分布 ,与其他群体进行比较 ,得出遗传距离 ,并了解它在法医学中的应用价值。　方法　采用多聚酶链式反应扩增这 6个基因座 ,采用非变性聚丙烯酰氨凝胶电泳银染显色分析。　结果　得出这 6个基因座在河南汉族人群中的基因频率 ,并计算得出杂合度、个体识别率、非父排除率 ,与其他群体比较得出进化距离。　结论　这 6个基因座有较高的杂合度 ,并且具有相对遗传稳定性 ,在人群中的分布符合Hardy -Weinberge平衡 ,有较高的法医学价值 ,可以应用于个体识别和亲权鉴定。     

3个Y-STR的复合扩增及其单倍型

目的 建立复合扩增Y-STR基因座的体系,获得广东汉族人的单倍型频率。方法 复合扩增DYS439、DYS437和DYS434三个基因座,用聚丙烯酰胺凝胶电泳银染法进行基因分型,检测广东汉族327名无关男性个体的单倍型。结果 3个基因座分别检出6个、4个和4个等位基因,共38种单倍型,其单倍型的个体识别率为0.8796。结论 Y-STR基因座复合扩增体系和建立的Y染色体STR数据库,在法医学鉴定中有应用意义。     

Genetic differentiation between and within Northern Native American language groups: an argument for the expansion of the Native American CODIS database

The National Research Council recommends that genetic differentiation among subgroups of ethnic samples be lower than 3% of the total genetic differentiation within the ethnic sample to be used for estimating reliable random match probabilities for forensic use. Native American samples in the United States’ Combined DNA Index System (CODIS) database represent four language families: Algonquian, Na-Dene, Eskimo-Aleut, and Salishan. However, a minimum of 27 Native American language families exists in the US, not including language isolates. Our goal was to ascertain whether genetic differences are correlated with language groupings and, if so, whether additional language families would provide a more accurate representation of current genetic diversity among tribal populations. The 21 short tandem repeat (STR) loci included in the Globalfiler® PCR Amplification Kit were used to characterize six indigenous language families, including three of the four represented in the CODIS database (i.e. Algonquian, Na-Dene, and Eskimo-Aleut), and two language isolates (Miwok and Seri) using major population genetic diversity metrics such as F statistics and Bayesian clustering analysis of genotype frequencies. Most of the genetic variation (97%) was found to be within language families instead of among them (3%). In contrast, when only the three of the four language families represented in both the CODIS database and the present study were considered, 4% of the genetic variation occurred among the language groups. Bayesian clustering resulted in a maximum posterior probability indicating three genetically distinct groups among the eight language families and isolates: (1) Eskimo, (2) Seri, and (3) all other language groups and isolates, thus confirming genetic subdivision among subgroups of the CODIS Native American database. This genetic structure indicates the need for an increased number of Native American populations based on language affiliation in the CODIS database as well as more robust sample sets for those language families.Supplemental data for this article is available online at https://doi.org/10.1080/20961790.2021.1963088 .     

Genetic polymorphism of 14 non-CODIS STR loci for forensic use in southeast China population

We investigated 14 polymorphic STR loci (D1S2142, D2S1360, D3S1545, D7S1517, D10S2325, D12S391, D13S1492, D14S306, D15S659, D16S3253, D18S1270, D19S253, D20S470, D21S1437) which are not included in the standard sets of forensic loci (CODIS) in a sample of 216 unrelated healthy southeast Chinese individuals. The studied loci were highly informative and did not show departures from Hardy-Weinberg equilibrium. The accumulated powers of discrimination and power of exclusion for the 14 loci were 99.9999999999 and 99.999998%, respectively. No linkage was observed between the 14 loci and the traditional set of STR markers included in commercially available kits (the AmpFLSTR IdentifilerTM 15 System loci). We thus considered the studied 14 STRs are informative and when necessary, can be used as the candidate genetic markers in the study and application in genetics and forensic practice.     

不同分型方法的STR分型差异

目的调查不同的STR分型系统之间分型的一致性。方法 10 0例不同个体的DNA样本分别用单位点聚丙烯酰胺凝胶银染法和PowerPlex16System试剂盒对 13个法医学常用STR位点进行基因分型 ,并比较两种不同分型系统间的分型结果。结果 1例样本在D8S1179位点出现了分型不一致的结果 :银染法的基因型为 12 / 14 ,而用PowerPlex16System试剂盒的分型则为 12 / 15。结论不同的STR分型系统可导致不同的基因分型     

Genetic variation at 15 autosomal microsatellite loci in the three highly endogamous tribal populations of Orissa, India

Although microsatellite diversity in autosomal chromosomes has been extensively described for many of the Indian populations, there is still a lacuna left on information about the genetic diversity of tribal populations. This paper reports the genetic data on the three tribal populations belonging to the Austroloid ethnic group from Orissa (Juang, Paroja and Saora). The 15 STR (D3S1358, THO1, D21S11, D18S51, PentaE, D5S818, D13S317, D7S820, D16S539, CSF1PO, PentaD, vWA, D8S1179, TPOX, FGA) polymorphism would help to accentuate the STR database for better understanding of population genetics and forensic applications. The microsatellites included in the system are found to be highly polymorphic, with the combined power of exclusion being greater than 0.999, in all the three investigated populations.