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1.
Toxicol Lett ; 319: 155-159, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31706005

RESUMO

Novel HepG2 cell clones 1A2 C2 and 1A2 C7 were independently generated by lentiviral transduction to functionally overexpress cytochrome P450 1A2 (CYP1A2). We found similar and stable CYP1A2 transcript and protein levels in both cell clones leading to specific enzyme activities of about 370 pmol paracetamol x min-1 x mg-1 protein analyzed by phenacetin conversion. Both clones showed dramatically increased sensitivity to the hepatotoxic compound aflatoxin B1 (EC50 < 100 nM) when compared to parental HepG2 cells (EC50∼5 µM). Thus, newly established cell lines are an appropriate tool to study metabolism and toxicity of substances depending on conversion by CYP1A2.


Assuntos
Citocromo P-450 CYP1A2/genética , Vetores Genéticos/genética , Hepatoblastoma/enzimologia , Hepatoblastoma/genética , Lentivirus/genética , Neoplasias Hepáticas/enzimologia , Neoplasias Hepáticas/genética , Aflatoxina B1/toxicidade , Linhagem Celular Tumoral , Citocromo P-450 CYP1A2/biossíntese , Impressões Digitais de DNA/métodos , Humanos , Fígado/metabolismo , Mycoplasma/química , Fenacetina/farmacocinética , Plasmídeos/genética
2.
J Forensic Sci ; 65(1): 209-213, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31433497

RESUMO

When using non-FTA cards in commercial multiplex STR kits for direct PCR, pretreatment steps with specific buffers are recommended. Here, we designed a rapid direct PCR method utilizing a non-FTA card, Oral Cell Sampling Kit, by omitting the pretreatment step involving Prep-n-Go™ Buffer, and it showed compatibility with the GlobalFiler™ Express PCR Amplification Kit, GlobalFiler™ PCR Amplification Kit, and PowerPlex® Fusion system. To optimize the PCR conditions, we tested the method with different final PCR volumes and cycles. Finally, we conducted a performance test using 50 Korean buccal samples and confirmed the high performance of the method, detecting more than 90% of the samples with full profiles when using GlobalFiler™ PCR Amplification Kit and PowerPlex® Fusion system at 29 cycles in a 10 µL final PCR volume. Thus, we report a simple direct PCR set-up to analyze reference samples collected using a non-FTA card manufactured in Korea.


Assuntos
Reação em Cadeia da Polimerase/métodos , Manejo de Espécimes/instrumentação , Impressões Digitais de DNA/métodos , Feminino , Humanos , Repetições de Microssatélites , Mucosa Bucal/química , República da Coreia , Manejo de Espécimes/métodos
3.
J Forensic Sci ; 65(1): 295-303, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30859587

RESUMO

A set of historic murders, known as the "Jack the Ripper murders," started in London in August 1888. The killer's identity has remained a mystery to date. Here, we describe the investigation of, to our knowledge, the only remaining physical evidence linked to these murders, recovered from one of the victims at the scene of the crime. We applied novel, minimally destructive techniques for sample recovery from forensically relevant stains on the evidence and separated single cells linked to the suspect, followed by phenotypic analysis. The mtDNA profiles of both the victim and the suspect matched the corresponding reference samples, fortifying the link of the evidence to the crime scene. Genomic DNA from single cells recovered from the evidence was amplified, and the phenotypic information acquired matched the only witness statement regarded as reliable. To our knowledge, this is the most advanced study to date regarding this case.


Assuntos
Vestuário , Impressões Digitais de DNA/métodos , Genética Forense/métodos , Homicídio/história , Manchas de Sangue , Vestuário/história , Vítimas de Crime , Criminosos , DNA Mitocondrial/genética , DNA Mitocondrial/isolamento & purificação , Fluorescência , História do Século XIX , Humanos , Raios Infravermelhos , Microdissecção e Captura a Laser , Reação em Cadeia da Polimerase , Polimorfismo de Nucleotídeo Único , Análise de Sequência de DNA , Análise de Célula Única , Reino Unido , Sequenciamento Completo do Genoma
4.
Fa Yi Xue Za Zhi ; 35(5): 519-524, 2019 Oct.
Artigo em Inglês, Chinês | MEDLINE | ID: mdl-31833283

RESUMO

Abstract: Genetic markers, such as single nucleotide polymorphism (SNP), insertion/deletion (InDel), were discovered and widely used with the development of whole genome sequencing and bioinformatics technology. The origin and genetic structure of the modern population had been gradually revealed from the perspective of genetics. The study on biogeographic ancestry inference in the field of forensic genetics emerged and developed rapidly, providing clues and scientific basis for the determination of investigation direction and for the narrow of the scope of investigation in the process of case investigation. This paper briefly reviews the research progress, inference methods and development trends of DNA ancestry inference technology.


Assuntos
Criminosos , Genética Forense/métodos , África , DNA/genética , Impressões Digitais de DNA/métodos , Genética Populacional , Humanos , Filogeografia , Polimorfismo de Nucleotídeo Único
5.
Forensic Sci Int ; 303: 109938, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31542402

RESUMO

Real forensic casework biological evidence can be found in a myriad of different conditions and presenting very distinct features, including key elements such as degradation levels, the nature of biological evidence, mixture presence, and surface or substrate deposition, among others. Technical protocols employed by forensic DNA analysts must consider such characteristics in order to improve the chances of successfully genotyping these materials. MPS has been used as a very useful tool for forensic sample processing and genetic profile generation. However, it is not completely clear how different features encountered with real forensic samples impact sequencing quality and, consequently, profile accuracy and reliability. In this context, the present study analyzes a set of 47 real forensic casework samples, obtained from semen, saliva, blood and epithelial evidence, as well as reference oral swabs, aiming to evaluate the impact of a sample's biological nature in profiling success. All DNA extracts from samples were standardized according to sample conditions, as assessed by traditional forensic profiling methods (real-time PCR quantitation and capillary electrophoresis-coupled STR fragment analysis). Samples were separated into groups according to their biological nature, and the resultant sequencing quality was evaluated through a series of well-established statistical tests, applied specifically to six different MPS quality metrics. The results showed that certain groups of samples, especially epithelial and (to a lesser extent) saliva samples, exhibited significantly lower quality in terms of some of the evaluated metrics. A number of reasons for such unexpected behavior are discussed. In addition, a series of calculations was performed to assess the weight of genetic evidence in Brazilian samples, and reflexes in data analysis and national allele frequency database construction are discussed. Overall, the results indicate that a unified national allele frequency database can be used nationwide. Besides this, MPS genetic profiles obtained from samples with particular biological origins may benefit from meticulous manual review, and visual inspection could be important as an additional step to avoid genotyping errors or misinterpretation, leading to more trustworthy and reliable results in real criminal forensic casework analysis.


Assuntos
Impressões Digitais de DNA/métodos , Sequenciamento de Nucleotídeos em Larga Escala , Polimorfismo de Nucleotídeo Único , Análise Química do Sangue , Brasil , Bases de Dados Genéticas , Eletroforese Capilar , Células Epiteliais , Frequência do Gene , Genética Populacional , Humanos , Repetições de Microssatélites , Reação em Cadeia da Polimerase em Tempo Real , Saliva , Sêmen , Análise de Sequência de DNA
6.
Forensic Sci Int Genet ; 43: 102140, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31536876

RESUMO

DNA mixture interpretation remains one of the major challenges in forensic DNA analysis. DNA mixture samples are inherently complex due to several factors including the variations in the quantity of DNA, the presence of non-allelic artifactual peaks and the presence of multiple contributors with variable levels of allele sharing. The Probabilistic Assessment for Contributor Estimation (PACE) is a fully continuous probabilistic machine learning-based method to predict the number of contributors (n) in a sample, and was previously developed for use with the Identifiler amplification kit. This system required manual preprocessing of data and was limited, exclusively, to samples amplified using said kit. This study introduces PACE™ v1.3.7 for use with both the GlobalFiler and PowerPlex Fusion 6c amplification kits. An automated artifact identification and management system has been added to accompany the rapid estimation of the number of donors in a given mixture. The artifact management module, when evaluated using previously unseen data, identified true allelic peaks and removed artifacts such as elevated baseline noise, stutter, and pull-up with accuracy over 93.5%. The systems yield the correct n classifications in over 90% of the samples, and demonstrate consistent accuracies as the number of donors and the overall mixture complexity increase. Misclassified samples generally exhibited high levels of allele sharing among donors, low DNA template amounts and high incidence of allelic dropout. This system offers a means for both artifact management and n estimation as well as a quantitative and reproducible method of assessing the quality of a profile.


Assuntos
Algoritmos , Artefatos , Impressões Digitais de DNA/métodos , DNA/genética , Aprendizado de Máquina , Alelos , Humanos , Modelos Estatísticos , Reação em Cadeia da Polimerase
7.
Forensic Sci Int Genet ; 43: 102150, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31476660

RESUMO

The number of contributors (NOC) to (complex) autosomal STR profiles cannot be determined with absolute certainty due to complicating factors such as allele sharing and allelic drop-out. The precision of NOC estimations can be improved by increasing the number of (highly polymorphic) markers, the use of massively parallel sequencing instead of capillary electrophoresis, and/or using more profile information than only the allele counts. In this study, we focussed on machine learning approaches in order to make maximum use of the profile information. To this end, a set of 590 PowerPlex® Fusion 6C profiles with one up to five contributors were generated from a total of 1174 different donors. This set varied for the template amount of DNA, mixture proportion, levels of allele sharing, allelic drop-out and degradation. The dataset contained labels with known NOC and was split into a training, test and hold-out set. The training set was used to optimize ten different algorithms with selection of profile characteristics. Per profile, over 250 characteristics, denoted 'features', were calculated. These features were based on allele counts, peak heights and allele frequencies. The features that were most related to the NOC were selected based on partial correlation using the training set. Next, the performance of each model (=combination of features plus algorithm) was examined using the test set. A random forest classifier with 19 features, denoted the 'RFC19-model' showed best performance and was selected for further validation. Results showed improved accuracy compared to the conventional maximum allele count approach and an in-house nC-tool based on the total allele count. The method is extremely fast and regarded useful for application in forensic casework.


Assuntos
Impressões Digitais de DNA/métodos , DNA/genética , Aprendizado de Máquina , Repetições de Microssatélites , Algoritmos , Alelos , Degradação Necrótica do DNA , Frequência do Gene , Humanos
8.
Forensic Sci Int ; 302: 109905, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31394460

RESUMO

Fingerprint detection and Short Tandem Repeat (STR) typing are two approaches used for identification of individuals. The main goal of forensic laboratories is the development of a standardized protocol to obtain an STR profile from latent fingerprints, by typing the DNA transferred onto touched objects. The results obtained in this field derive from studies conducted under controlled laboratory conditions. Here, for the first time, we report two different profiles obtained by DNA purified by latent fingerprints enhanced with dactyloscopic powders at a crime scene, 14 years previously. DNA extraction phase was optimized to improve removal of powder and automatically conducted. Despite the low concentration of purified DNA, it was not degraded. Even if quality of the profile is influenced by several factors such as the method of acquisition and storage conditions of the fingerprint, results obtained are adequately informative and could be uploaded to the CODIS database.


Assuntos
Impressões Digitais de DNA/métodos , DNA/isolamento & purificação , Dermatoglifia , Repetições de Microssatélites , Eletroforese Capilar , Humanos , Pós , Reação em Cadeia da Polimerase em Tempo Real , Análise de Regressão , Manejo de Espécimes/métodos
9.
Biol Direct ; 14(1): 13, 2019 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-31420049

RESUMO

BACKGROUND: Research has found that human associated microbial communities play a role in homeostasis and the disruption of these communities may be important in an array of medical conditions. However outside of the human body many of these communities remain poorly studied. The Metagenomics and Metadesign of the Subways and Urban Biomes (MetaSUB) International Consortium is characterizing the microbiomes of urban environments with the aim to improve design of mass transit systems. As part of the CAMDA 2018 MetaSUB Forensics Challenge 311 city microbiome samples were provided to create urban microbial fingerprints, as well as a further 3 mystery datasets for validation. RESULTS: MetaSUB samples were clustered using t-SNE in an unsupervised fashion to almost discrete groups, which upon inspection represented city of origin. Based on this clustering, geographically close metropolitan areas appear to display similar microbial profiles such as those of Auckland and Hamilton. Mystery unlabeled samples were provided part of the challenge. A random forest classifier built on the initial dataset of 311 samples was capable of correctly classifying 83.3% of the mystery samples to their city of origin. Random Forest analyses also identified features with the highest discriminatory power, ranking bacterial species such as Campylobacter jejuni and Staphylococcus argenteus as highly predictive of city of origin. The surface from which the sample was collected displayed little detectable impact on the microbial profiles in the data generated here. The proportion of reads classified per sample varied greatly and so de-novo assembly was applied to recover genomic fragments representing organisms not captured in reference databases. CONCLUSIONS: Current methods can differentiate urban microbiome profiles from each other with relative ease. De-novo assembly indicated that the MetaSUB metagenomic data contains adequate depth to recover metagenomic assembled genomes and that current databases are not sufficient to fully characterize urban microbiomes. Profiles found here indicate there may be a relationship between geographical distance between areas and the urban microbiome composition although this will need further research. The impact of these different profiles on public health is currently unknown but the MetaSUB consortium is uniquely suited to evaluate these and provide a roadmap for the inclusion of urban microbiome information for city planning and public health policy. REVIEWERS: This article was reviewed by Dimitar Vassilev, Eran Elhaik and Chengsheng Zhu.


Assuntos
Impressões Digitais de DNA/métodos , Aprendizado de Máquina , Metagenoma/genética , Metagenômica/métodos , Microbiota/genética , Cidades
10.
Forensic Sci Int Genet ; 43: 102148, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31446344

RESUMO

Hair is an evidentiary sample that typically does not provide sufficient nuclear DNA for forensic analysis. Therefore, state-of-the-art forensic examination for hair samples include subjective microscopic evaluation, mitochondrial DNA (mtDNA) analysis, and more recently, proteomic genotyping that uses protein variation in the form of genetically variant peptides (GVPs) to infer single nucleotide polymorphism (SNP) alleles. Since many cases involve limited sample amounts (approximately 2 cm or less), any additional destructive testing (besides mtDNA) would be excluded. If a mtDNA-compatible protein extraction workflow could be developed, GVPs would provide additional forensic value without sacrificing any portion of the original hair sample. Here, we demonstrate an optimized method that can be used to obtain both whole genome mtDNA and putative GVP profiles from a single limited hair sample. The method involves urea-based extraction of proteins from hair, followed by buffer exchange and protease digestion. Peptides are eluted through a 30 kDa membrane and analyzed using traditional proteomic techniques. DNA is subsequently extracted from the filter and analyzed using whole mt-genome analysis. The method was verified with a range of hair sample types (head, pubic, and arm hair) from a diverse cohort of 22 individuals. Specifically, putative GVP profiles and mtDNA haplotypes concordant with buccal swab samples from the same donor were obtained from 22 individuals. Further, the utility of the method was verified across two different laboratories. The method is applicable for proteomic-based GVP analysis and mt-genome analysis for forensic research applications.


Assuntos
Impressões Digitais de DNA/métodos , DNA Mitocondrial/genética , Cabelo/química , Peptídeos/genética , Adulto , Feminino , Genoma Mitocondrial , Técnicas de Genotipagem , Haplótipos , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Espectrometria de Massas , Pessoa de Meia-Idade , Peptídeos/análise , Reação em Cadeia da Polimerase , Polimorfismo de Nucleotídeo Único , Proteômica , Análise de Sequência de DNA , Fluxo de Trabalho , Adulto Jovem
11.
Forensic Sci Int Genet ; 42: 198-202, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31376647

RESUMO

Evidence of sexual aggression may be obtained from superabsorbent polymer (SAP) sanitary pads, which are used by forensic laboratories for semen evaluation. Semen can be extracted from their upper layers, which are free of SAPs. However, our previous results showed a need to optimize the protocol for semen analysis by considering its extraction from the lower core, often composed of sodium polyacrylate SAPs. SAPs generate a hydrogel, which traps the cellular components, hindering the possibility of obtaining cells and hence their genetic material. Simple filtration has been tried previously, but further maximization by application of a treatment has never been attempted. In this paper, we compare both chemical and physical shredding treatments for maximizing gel-trapped sperm and male cell DNA recaptures from hygienic superabsorbent substrates in sanitary pads, panty-liners or diapers. Our findings suggest that the lower core should be treated to induce a dewaterisation of the SAP hydrogels in order to maximize the extraction of bodily fluids.


Assuntos
Impressões Digitais de DNA/métodos , Produtos de Higiene Menstrual , Sêmen , Delitos Sexuais , Manejo de Espécimes/métodos , DNA/isolamento & purificação , Células Epiteliais , Feminino , Humanos , Indicadores e Reagentes , Masculino , Repetições de Microssatélites , Reação em Cadeia da Polimerase , Polímeros , Manejo de Espécimes/instrumentação , Espermatozoides , Vagina/citologia
12.
J Forensic Sci ; 64(6): 1646-1657, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31343733

RESUMO

Between 1990 and 2018, the Defense POW/MIA Accounting Agency submitted 2177 cranial elements and 1565 teeth to the Armed Forces Medical Examiner System-Armed Forces DNA Identification Laboratory for DNA testing. In an effort to identify missing United States service members, materials were recovered from wartime losses inclusive of World War II, the Korean War, and Southeast Asia. Using four different DNA extraction protocols, DNA testing was performed using mitochondrial DNA Sanger sequencing, modified AmpFlSTR® Yfiler™, AmpFlSTR® MiniFiler™, PowerPlex® Fusion, or Next Generation Sequencing. This paper aims to provide optimal strategies for the DNA testing of skeletonized cranial materials. Cranial elements produced the most consistent results in Sanger sequencing using an organic purification; however, teeth were most successful for the same platform with an inorganic purification. The inverse is true for STR testing of cranial bones. Of the cranial elements, the temporal provided the most consistent results.


Assuntos
Impressões Digitais de DNA/métodos , DNA Mitocondrial/isolamento & purificação , Crânio/química , Dente/química , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Repetições de Microssatélites , Militares , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Manejo de Espécimes/métodos
13.
Medicina (Kaunas) ; 55(7)2019 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-31311196

RESUMO

Background and objectives. Human papillomavirus (HPV) is the most commonly sexually transmitted infection. Recent evidence suggests that an HPV infection may affect fertility. The aim of the study was to determine the prevalence of HPV infections among couples undergoing in vitro fertilization (IVF) and to identify their awareness of HPV. Material and Methods. A total of 200 samples were collected from couples who received IVF treatment during 2017-2018 in Vilnius University Hospital Santaros Klinikos (VUH SK) Santaros Fertility Centre (SFC). For HPV detection, cervical swabs from women and sperm samples from men were taken and a real time polymerase chain reaction (RT-PCR) was used for the identification of 14 high-risk HPV types. Sperm parameters were evaluated according to World Health Organization (WHO) recommendations for 2010. Research subjects answered an anonymous questionnaire to ascertain their knowledge of HPV. Results. After testing of HPV in couples undergoing IVF, it was found that 33 out of 100 couples (33%) were HPV positive. Of these, 19% of women (19/100) and 20% of men (20/100) tested positive. Using Fisher's exact test, a statistically significant difference was found between HPV infections and abnormal sperm quality parameters (p = 0.023). Conclusions. HPV may have an impact in spermatogenesis, because an HPV infection was more frequently detected in men with abnormal sperm parameters. High-risk HPV 52 was the most common genotype among couples undergoing IVF treatment.


Assuntos
Infertilidade/etiologia , Infecções por Papillomavirus/complicações , Adulto , Impressões Digitais de DNA/métodos , Feminino , Humanos , Infertilidade/epidemiologia , Infertilidade/genética , Lituânia/epidemiologia , Masculino , Pessoa de Meia-Idade , Papillomaviridae/genética , Infecções por Papillomavirus/epidemiologia , Infecções por Papillomavirus/genética , Prevalência , Espermatozoides/microbiologia , Esfregaço Vaginal/métodos
14.
Forensic Sci Int Genet ; 41: 168-176, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31153002

RESUMO

Bombing accounts for the largest share of terrorist incidents worldwide. Most involve an improvised explosive device (IED): a bomb made from household items. Touch DNA may be left on parts of an IED during assembly. However, an IED conflagration degrades DNA, and there has never been a way to locate where touch DNA may remain. To solve this problem, we combined the use of fluorescent dye to locate latent DNA and direct PCR to improve STR profiles of DNA obtained from IEDs. Six fluorescent DNA-binding dyes were evaluated at various concentrations for the purpose of staining latent DNA. SYBR® Green I and Diamond™ Nucleic Acid dye were able to visualize touch DNA on IED substrates. Inhibition studies with extracted DNA and touch DNA using both dyes revealed that Diamond™ dye inhibited direct STR amplification, while SYBR® Green I did not. Stability studies at three temperatures showed optimum performance of SYBR® Green I up to 24 h after formulation. As such, only SYBR® Green I was further used to develop a "visualized-direct PCR" method. Using the conventional approach and the novel "visualized-direct PCR" approach in a single-blind investigation of mock IED evidence, the "visualized-direct PCR" approach had a 98.6% chance of obtaining more alleles (95% highest density interval (HDI): 0.7 to 10.0 alleles). A decrease in non-donor's alleles (mixed profiles) was also observed. The developed approach has the potential to revolutionize the process of STR typing from touch DNA.


Assuntos
Bombas (Dispositivos Explosivos) , Impressões Digitais de DNA/métodos , Corantes Fluorescentes , Repetições de Microssatélites , Reação em Cadeia da Polimerase/métodos , Tato , Alelos , Humanos , Compostos Orgânicos
15.
Forensic Sci Int ; 301: 101-106, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31153987

RESUMO

In forensic genetics, the analysis of DNA in biological samples is a valuable tool for personal identification. There is an increasing demand in analyzing of the mixed DNA which may provide insightful investigative instructions. With the continuous effort for the improvement of individual identification, complicated mixed stains represent a growing fraction of the samples processed by forensic laboratories. Recent technological advances have enabled quantitative analysis of DNA mixture and emerging sequencing approaches to decipher the complicated DNA mixture. Here, we describe the use of different genetic markers, typing approaches and analytical methods in mixture analysis, and how useful information can be obtained from complicated DNA mixture.


Assuntos
Impressões Digitais de DNA/métodos , DNA/análise , Genética Forense/métodos , Marcadores Genéticos , Haplótipos , Humanos , Funções Verossimilhança , Repetições de Microssatélites , Polimorfismo de Nucleotídeo Único , Análise de Sequência de DNA
16.
Leg Med (Tokyo) ; 39: 45-48, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31207384

RESUMO

Genetic polymorphism of 21 autosomal STR loci included in Goldeneye™ DNA ID 22NC Kit (D19S253, D6S477, D22GATA198B05, D15S659, D8S1132, D3S1358, D3S3045, D14S608, D17S1290, D3S1744, D2S411, D18S535, D13S325, D7S1517, D10S1435, D11S2368, D4S2366, D1S1656, D7S3048, D10S1248 and D5S2500) was studied in 297 unrelated She individuals. Allele frequencies and forensic efficiency parameters such as, observed and expected heterozygosity (Ho & He), matching probability (MP), power of discrimination (PD), polymorphism information content (PIC), power of exclusion (PE), typical paternity index (TPI) was calculated for the loci. The combined PD and PE for all 21 STR loci were calculated to be 0.999999999999999999999999324094 and 0.999999996182425, respectively. The dataset indicated the usefulness of these loci in personal identification, parentage testing and complex kinship analysis in She population. The MDS plots and neighbor-joining tree were constructed based on pair-wise Nei's genetic distance by comparing allele frequency data for the 21 loci with ten other populations. The analysis showed that She population lies closer to a clade consisting Southern Han, Hainan Han and Li populations.


Assuntos
Cromossomos Humanos Par 21/genética , Impressões Digitais de DNA/métodos , Genética Forense/métodos , Loci Gênicos/genética , Genética Populacional/métodos , Repetições de Microssatélites/genética , Grupo com Ancestrais do Continente Asiático/genética , China , Frequência do Gene , Humanos
17.
Forensic Sci Int Genet ; 42: 49-55, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31252251

RESUMO

This study reports Short Tandem Repeat (STR) sequence-based allele data from 496 Spanish individuals across 31 autosomal STR (auSTR) loci included in the Precision ID GlobalFiler™ NGS STR Panel v2: D12S391, D13S317, D8S1179, D21S11, D3S1358, D5S818, D1S1656, D2S1338, vWA, D2S441, D5S2800, D7S820, D16S539, D6S474, D12ATA63, D4S2408, D6S1043, D19S433, D14S1434, CSF1PO, D10S1248, D18S51, D1S1677, D22S1045, D2S1776, D3S4529, FGA, Penta D, Penta E, TH01 and TPOX. The sequence of each allele was aligned to the reference sequence GRCh37 (hg19) and formatted according to the guidance of the International Society for Forensic Genetics. A subset of 221 samples was evaluated for testing concordance with allele calls derived from CE-based analysis using PowerPlex Fusion 6C, and there was 99.95% allele concordance. Twenty-five out of 31 auSTR loci showed an increased number of alleles due to repeat region sequence variation and/or single nucleotide polymorphisms (SNP) residing in the flanking regions. A total of 18 loci showed increased observed heterozygosity due to sequence variation; the loci exhibiting the greatest increase were: D13S317 (12% points), D5S818 (10% points), D8S1179 (7% points), D3S1358 (7% points), and D21S11 (6% points). The combined match probability decreased from 2.022E-24 (length-based data) to 1.042E-27 (sequence-based data) for the 20 CODIS core STR loci. The combined match probability (sequence-based data) for the 31 STR loci studied was 4.777E-40. The combined typical paternity index increased from 1.118E + 12 to 8.179E + 13 using length and sequence-based data, respectively. This Spanish population study performed in the framework of the EU-funded DNASEQEX project is expected to provide STR sequence-based allele frequencies for forensic casework and support implementation of massively parallel sequencing (MPS) technology in forensic laboratories.


Assuntos
Impressões Digitais de DNA/métodos , Eletroforese Capilar , Sequenciamento de Nucleotídeos em Larga Escala , Repetições de Microssatélites , Análise de Sequência de DNA , Frequência do Gene , Genética Populacional , Humanos , Polimorfismo de Nucleotídeo Único , Espanha
18.
Yakugaku Zasshi ; 139(5): 725-730, 2019.
Artigo em Japonês | MEDLINE | ID: mdl-31061341

RESUMO

As criminal cases have become more complicated, Japan's law enforcement officials are promoting the use of more sophisticated technologies, such as DNA analysis, in the course of criminal investigations in order to verify facts with objective evidence. The primary DNA analysis method employed by law enforcement officials is short tandem repeat (STR) analysis, a method for identifying individuals utilizing individual differences in the number of repeat units of characteristic DNA sequences. Presently, STR analysis can discriminate between individuals with the probability of one in approximately 4.7 trillion, even when the DNA profile is the most common type among the Japanese population. In every prefectural police department, members of criminal investigation laboratories, who were trained and certified by the Training Center of Forensic Science at the National Research Institute of Police Science, perform STR analysis. Forensic DNA analysis plays an important role not only in criminal investigations but also following large-scale disasters, to aid in individual identification. The accuracy of DNA typing is increasing with the availability of STR typing kits that can examine more loci than conventional kits. However, it remains difficult for DNA analysis to identify individuals with only small amounts of samples, old samples, or mixed samples. New methods for handling these problematic samples are required. Here, we review current investigative techniques and challenges of DNA analysis, and focus on the latest research for solutions to these challenges.


Assuntos
Crime , Impressões Digitais de DNA/métodos , Antropologia Forense/métodos , Repetições de Microssatélites/genética , Análise de Sequência de DNA/métodos , Humanos , Japão
19.
Electrophoresis ; 40(14): 1824-1829, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31106865

RESUMO

A variety of evidences are found at crime scenes. Fingerprint and DNA evidences are especially important in the process of identifying personal sources. Among evidences found at crime scenes, cigarette butts are important because they might contain both fingerprints and DNA. In this study, latent fingerprints were detected in cigarette butts using 1,8-diazafluoren-9-one (DFO) and 1,2-Indanedione/zinc chloride (1,2-IND/Zn). Next, DNA extraction and real-time qPCR were performed to quantify and identify the DNA present. Short tandem repeat (STR) profiling was also performed. The results showed that the quantity of DNA recovered was decreased by 16% in DFO-treated cigarettes and by 27% in 1,2-IND/Zn-treated cigarettes when compared to untreated controls. When the STR profiling results were compared with those of the control sample, DFO, and 1,2-IND/Zn reagent-treated DNA samples showed individualized genotyping at several loci. Results of this study showed that when cigarette butts were found, DFO and 1,2-IND/Zn reagents could be used for DNA profiling after fingerprint identification. However, the effect of DFO on STR profiling was less than that of 1,2-IND/Zn. Therefore, we recommend the use of DFO for fingerprinting cigarette butts if further DNA processing is planned.


Assuntos
Compostos Aza/química , Impressões Digitais de DNA/métodos , DNA/análise , Saliva/química , Cloretos/química , Eletroforese Capilar , Corantes Fluorescentes , Medicina Legal/métodos , Humanos , Indanos/química , Reação em Cadeia da Polimerase em Tempo Real , Espectrometria de Fluorescência , Compostos de Zinco/química
20.
Forensic Sci Int Genet ; 41: 107-119, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31071519

RESUMO

The deconvolution of DNA mixtures has gathered the attention of forensic DNA scientists for over two decades. To enhance mixture deconvolution capabilities, a new generation of sensitive DNA-typing approaches has been recently proposed. In this review, we describe novel, forensically relevant multi-SNP loci (i.e., microhaplotypes or microhaps), compound markers (i.e., DIP-STRs, SNP-STRs and DIP-SNPs) and lineage markers (i.e., rapidly mutating Y chromosome STRs) that improve the deconvolution of two and more than two-person mixtures typed using conventional STR, binary and non-binary loci. We explore the features and applications of these emerging molecular biomarkers with respect to their ability to forensically detect same-or-opposite sex donors. Finally, we discuss the impact of initial massively parallel sequencing (MPS) investigations of STR, microhaplotype and SNP/indel assays for DNA mixture profiling.


Assuntos
Impressões Digitais de DNA/métodos , DNA/genética , Marcadores Genéticos , Cromossomos Humanos Y , Eletroforese Capilar , Haplótipos , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Mutação INDEL , Repetições de Microssatélites , Reação em Cadeia da Polimerase , Polimorfismo de Nucleotídeo Único , Análise de Sequência de DNA
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