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1.
Mem Inst Oswaldo Cruz ; 114: e190260, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31644705

RESUMO

BACKGROUND: Sporotrichosis is a subcutaneous mycosis caused by dimorphic pathogenic fungi belonging to the Sporothrix genus. Pathogenic Sporothrix species typically produce melanin, which is known to be a virulence factor. OBJECTIVES: The aim of this study was to perform phenotypic, genotypic, and virulence analyses of two distinct Sporothrix brasiliensis strains isolated from the same lesion on a patient from Rio de Janeiro. METHODS AND FINDINGS: Genotypic analyses by partial sequencing of the calmodulin, ß-tubulin, and chitin synthase genes, as well as polymerase chain reaction (PCR)-fingerprinting by T3B, M13, and GACA, showed that the isolates were very similar but not identical. Both isolates had similar phenotypic characteristics and effectively produced melanin in their yeast forms, accounting for their ability of causing disease in a murine sporotrichosis model. Remarkably, isolate B was albino in its environmental form but caused more severe disease than the pigmented A isolate. CONCLUSIONS: These findings indicate that the patient was infected by two genetically and biologically distinct S. brasiliensis that vary in their production of melanin in their environmental forms. The results underscore the importance of characterizing phenotypically different isolates found in the same clinical specimen or patient.


Assuntos
Antifúngicos/farmacologia , Sporothrix/patogenicidade , Esporotricose/patologia , Esporotricose/virologia , Animais , Impressões Digitais de DNA , Modelos Animais de Doenças , Genótipo , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Testes de Sensibilidade Microbiana , Fenótipo , Reação em Cadeia da Polimerase , Sporothrix/efeitos dos fármacos , Sporothrix/genética , Virulência
2.
Gene ; 720: 144078, 2019 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-31473321

RESUMO

Short tandem repeats (STRs) are a widely utilized tool in forensic applications, the latter of which range from human identification and paternity testing to population analysis. The GlobalFiler STR loci, which includes 21 autosomal STRS, were analyzed in the Chechen subpopulation of Jordan. Whole blood samples were withdrawn from 159 Jordanian Chechen individuals, and genomic DNA was extracted from each sample. The GlobalFiler™ kit PCR Amplification Kit amplified and analyzed the STR loci on the 3130xl Genetic Analyzer using GeneMapper ID-X software. The combined match probability for the 21 autosomal STR loci was calculated to be 1.06 × 10-24, a number that is highly discriminatory and informative. The SE33 (0.983) and TPOX (0.806) loci exhibited the highest and lowest powers of discrimination, respectively. Conclusively, the current study indicates that the GlobalFiler loci have a high utility in the Jordanian Chechen population, possibly paving the way for the future establishment of a reference population database in Jordan.


Assuntos
DNA/análise , DNA/genética , Grupos Étnicos/genética , Genética Forense/estatística & dados numéricos , Genética Populacional , Repetições de Microssatélites , Polimorfismo Genético , Impressões Digitais de DNA , Feminino , Frequência do Gene , Loci Gênicos , Humanos , Masculino
3.
J Forensic Leg Med ; 67: 24-27, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31377682

RESUMO

BACKGROUND: A mass grave is any site that containing two or more associated corpses, at random or on purpose placed, of people who have died as a result of extra-judicial or random executions, not including those people who have died from armed confrontations or known major catastrophes. CASE PRESENTATION: The purpose of this paper is to explain how to reconstruct a biological profile of decomposed or skeletonized bodies and clarify the efforts done by the Libyan scientist after 2011 revolution and to set a reference for other researcher. The alleged location of the grave, as well as the alleged number and identities of the persons buried in the grave were obtained exclusively from witnesses' and relatives' testimonies. CONCLUSION: As the testimonies said, the grave was located at the alleged location and seven skeletons were exhumed. Also, the osteological and DNA study made investigators to identify the exhumed skeletons. And the dental analysis support the identification of a seven man alleged to have been buried in the grave, 7 victims were discovered.


Assuntos
Osso e Ossos/patologia , Impressões Digitais de DNA , Exumação , Adulto , Determinação da Idade pelo Esqueleto , Sepultamento , DNA/isolamento & purificação , Antropologia Forense , Odontologia Legal , Humanos , Líbia , Masculino , Repetições de Microssatélites , Determinação do Sexo pelo Esqueleto , Dente/química , Guerra
4.
Georgian Med News ; (290): 157-163, 2019 May.
Artigo em Russo | MEDLINE | ID: mdl-31322535

RESUMO

The purpose of the study is to analyze the effectiveness of using the capabilities of a forensic molecular genetic examination to solve the problems of criminal proceeding in Ukraine. The authors analyze the regulatory acts governing criminal procedure and forensic activities in Ukraine in context of identifying legal problems of using molecular genetic expertise in criminal proceedings. 568 sentences handed down by the courts of Ukraine in 2014-2018 in criminal cases of homicides (250 sentences), robberies (250 sentences) and road traffic accidents (68 sentences), during the investigation of which specified examinations were appointed, as well as 123 conclusions of molecular genetic examinations conducted in 2018. A summary of studied materials in context of solving the problems of establishing DNA profiles by experts, the correctness and completeness of posed questions and the conclusions made by an expert, the value of the results obtained for the defendant to establish whether the defendant was guilty or not guilty, the reasons why the expert task was impossible are conducted. It is established that the effectiveness of the application of the results of molecular genetic expertise in the criminal proceedings of Ukraine can be improved providing that a number of legal, organizational, methodological and procedural problems are solved. the need for legislative regulation of the procedure for the selection of biological samples from various categories of persons and their inclusion in automated databases of DNA profiles; substantial replenishment of these bases; establishment international cooperation of domestic bases with foreign information systems are indicated. Unification is required on a uniform methodological basis and toolkit of a system of expert institutions for carrying out molecular genetic examinations, training of employees of investigative and operational-search units to work at the crime scene with traces of biological origin. Further development and improvement of DNA analysis methods should include the development of new and improvement of existing methods of molecular genetic examination, especially in the study of mixed and micro-amounts of biological traces. It is necessary to develop criteria (standards) for assessing the coincidence of DNA profiles so that not only could experts determine the likelihood of such a coincidence, but also formulate conclusions about identity that are available to the court.


Assuntos
Criminosos , Impressões Digitais de DNA , Patologia Legal , Criminosos/legislação & jurisprudência , Humanos , Ucrânia
5.
Fa Yi Xue Za Zhi ; 35(3): 319-323, 2019 Jun.
Artigo em Inglês, Chinês | MEDLINE | ID: mdl-31282628

RESUMO

Abstract: Objective To investigate the application of the comprehensive use of multiple genetic markers in full and half sibling relationship testing through the identification of a case of suspected sibling relationship. Methods Genomic DNA were extracted from bloodstain samples from 4 subjects (ZHANG-1, ZHANG-2, male; ZHANG-3, ZHANG-4, female). Autosomal STR loci, X-STR, Y-STR loci and polymorphisms of mtDNA HV-Ⅰ and Ⅱwere genotyped by EX20 STR kit, X19 kit, Data Y24 STR kit, and Sanger sequencing, respectively. Results According to autosomal STR based IBS scoring results, full sibling relationships were indicated among ZHANG-2, ZHANG-3 and ZHANG-4, but those were not indicated between ZHANG-1 and ZHANG-2 or ZHANG-3 or ZHANG-4. According to autosomal STR based FSI and HSI, with ITO method and discriminant function method, full sibling relationships among ZHANG-2, ZHANG-3 and ZHANG-4 were indicated, and half sibling relationships between ZHANG-1 and ZHANG-2 or ZHANG-3 or ZHANG-4 were also indicated. X-STR and mtDNA sequencing results showed that all the 4 samples came from a same maternal line, and Y-STR results showed that ZHANG-1 and ZHANG-2 did not come from a same paternal line, which supported the half sibling relationship between ZHANG-1 and ZHANG-2 or ZHANG-3 or ZHANG-4, verified by parental genotype reconstruction based on autosomal STR genotyping. Conclusion For the identification of sibling relationships, it is effective to have reliable results with the mutual verification and support of multiple genetic markers (autosomal STR, sex chromosomal STR and mtDNA sequence) and calculations (IBS, ITO, discriminant function method and family reconstruction).


Assuntos
Genética Forense , Irmãos , Alelos , Cromossomos Humanos Y , Impressões Digitais de DNA , Feminino , Marcadores Genéticos , Genótipo , Humanos , Masculino , Repetições de Microssatélites
6.
Fa Yi Xue Za Zhi ; 35(3): 324-327, 2019 Jun.
Artigo em Inglês, Chinês | MEDLINE | ID: mdl-31282629

RESUMO

Abstract: Objective To perform the separation and confirmation of mixed semen stains with immunological test method, and find a more effective method for the detection of mixed semen stains. Methods The semens of three volunteers were mixed. The mixed semen stains were processed and tested with prostate-specific antigen (PSA) colloidal gold immunoassay strip method, immunomagnetic beads method and laser capture microdissection, respectively. Statistics of the results of STR were gathered and compared with those of a single semen stain. Results After PSA colloidal gold immunoassay strip method testing, the samples showed a purplish red line in the test area and the control area. The results obtained with the immunomagnetic beads method showed a more complete and effective short tandem repeat (STR) sequence. The mixed semen stains were processed with laser capture microdissection and low volume amplified. The results were summarized and superimposed to obtain a complete single typing, which matched the single semen stain typing, with a typing success rate of 84.00%. Single suspect Y-STR typing was obtained with the application of the method above in actual cases, which provided evidence basis for rapid solving of the case. Conclusion The combination of PSA colloidal gold immunoassay strip method, immunomagnetic beads method and laser capture microdissection can be used to separate and confirm the mixed semen stains.


Assuntos
Sêmen , Corantes , Impressões Digitais de DNA , Medicina Legal , Humanos , Testes Imunológicos , Masculino , Repetições de Microssatélites
7.
J Microbiol Biotechnol ; 29(7): 1144-1154, 2019 Jul 28.
Artigo em Inglês | MEDLINE | ID: mdl-31288301

RESUMO

There have been several studies regarding lichen-associated bacteria obtained from diverse environments. Our screening process identified 49 bacterial species in two lichens from the Himalayas: 17 species of Actinobacteria, 19 species of Firmicutes, and 13 species of Proteobacteria. We discovered five types of strong antimicrobial agent-producing bacteria. Although some strains exhibited weak antimicrobial activity, NP088, NP131, NP132, NP134, and NP160 exhibited strong antimicrobial activity against all multidrug-resistant strains. Polyketide synthase (PKS) fingerprinting revealed results for 69 of 148 strains; these had similar genes, such as fatty acid-related PKS, adenylation domain genes, PfaA, and PksD. Although the association between antimicrobial activity and the PKS fingerprinting results is poorly resolved, NP160 had six types of PKS fingerprinting genes, as well as strong antimicrobial activity. Therefore, we sequenced the draft genome of strain NP160, and predicted its secondary metabolism using antiSMASH version 4.2. NP160 had 46 clusters and was predicted to produce similar secondary metabolites with similarities of 5-100%. Although NP160 had 100% similarity with the alkylresorcinol biosynthetic gene cluster, our results showed low similarity with existing members of this biosynthetic gene cluster, and most have not yet been revealed. In conclusion, we expect that lichen-associated bacteria from the Himalayas can produce new secondary metabolites, and we found several secondary metabolite-related biosynthetic gene clusters to support this hypothesis.


Assuntos
Anti-Infecciosos/metabolismo , Genoma Bacteriano/genética , Líquens/microbiologia , Streptomyces/genética , Streptomyces/metabolismo , Anti-Infecciosos/farmacologia , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Sequência de Bases , Impressões Digitais de DNA , DNA Bacteriano/genética , Testes de Sensibilidade Microbiana , Família Multigênica , Filogenia , Policetídeo Sintases/genética , RNA Ribossômico 16S/genética , Metabolismo Secundário/genética , Análise de Sequência de DNA
8.
Anthropol Anz ; 76(4): 333-351, 2019 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-31322643

RESUMO

Skeletal remains are among the most difficult types of samples encountered in forensic DNA casework and historical investigations due to prolonged exposure to environmental insults. DNA extracted from bone often is degraded, in low quantities, and contains co-purified inhibitors from the surrounding soil and/or burial vault material. When sexually dimorphic skeletal elements are not recovered, determining the sex of a decedent can be challenging. With unidentified human skeletal remains, genetic data often are evaluated in concert with anthropological analyses, as well as other types of metadata, to improve confidence in making associations or for positive identifications. This study evaluated a multi-faceted molecular genetic approach to increasing the amount of data that can be recovered from degraded skeletal remains. Results demonstrate that using a newer-generation multiplex (GlobalFiler™) with an expanded set of highly discriminatory DNA markers - combined with co-amplification of three different sex-determining loci, one additional PCR cycle, and testing multiple cuttings from the same bone or multiple regions within a skeleton - can improve reliability and accuracy in skeletal remains identifications by providing data concordance.


Assuntos
Restos Mortais , Impressões Digitais de DNA , DNA , Antropologia , Consenso , DNA/isolamento & purificação , Humanos , Repetições de Microssatélites , Reprodutibilidade dos Testes
9.
J Forensic Leg Med ; 66: 117-119, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31299483

RESUMO

DNA can be useful corroborative evidence in establishing familial relationship in immigration cases. Presently, there is no specific law in the UK regulating the use of DNA in this domain. This has led to inconsistencies in policy guidance and the rejection of some immigrant applications solely or partly due to a lack of DNA evidence. This commentary draws on the DNA regulatory regime in law enforcement to make a case for a specific DNA immigration law to protect individual rights, assure fairness and trust in the treatment of applicants. In addition to a specific law, consistency in operations should be ensured by developing a central point of contact for guidance including a central IT system, and a custodian of the DNA application process. Further, a single code of practice and conduct is proposed to ensure that guidance products are in line with the law and practice. An independent multi-stakeholder board is also recommended to ensure that policies are representative of the views of applicants and their relatives; policy officers and operational staff; and policymakers and the public.


Assuntos
Impressões Digitais de DNA/legislação & jurisprudência , Emigrantes e Imigrantes/legislação & jurisprudência , Emigração e Imigração/legislação & jurisprudência , Humanos , Aplicação da Lei , Medidas de Segurança/legislação & jurisprudência , Reino Unido
10.
J Forensic Leg Med ; 66: 155-161, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31306915

RESUMO

The simultaneous localisation and globalisation of 'terrorist threats' and cross-border criminality have led to increased expansion of surveillance activities and greater cross-border police and judicial cooperation, placing a greater priority on these activities within the political agenda of the EU. In this scenario, the expansion of technological systems for surveillance and monitoring, and the large-scale exchange of citizens' personal data play a pivotal role in the "fight against crime". This paper explores the multiplicity of data protection regimes in different EU Member States within the framework of the Prüm system. While EU regulations establish minimum standards for personal data flows at the transnational level, local and domestic practices are extremely heterogeneous. Based on analysis of 37 interviews conducted with professionals involved in the automated exchange of forensic genetic profiles, this paper provides empirical data that highlights the tensions between the local and the global within DNA data exchanges across the EU. These tensions relate to differentiated sociotechnical imaginaries regarding the protection of personal data flowing between Member-States. In sum, this paper analyses the potential threats to human rights created by the exchange of personal data with regards to issues of privacy and data protection.


Assuntos
Segurança Computacional/legislação & jurisprudência , Bases de Dados de Ácidos Nucleicos , Disseminação de Informação/legislação & jurisprudência , Cooperação Internacional , Privacidade/legislação & jurisprudência , Crime/prevenção & controle , Impressões Digitais de DNA/legislação & jurisprudência , Dermatoglifia , União Europeia , Humanos , Terrorismo/prevenção & controle
11.
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
12.
Forensic Sci Int ; 301: 107-117, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31153988

RESUMO

In the last year direct-to-consumer (DTC) genetic genealogy databases have been used to identify suspects and missing persons in over fifty cold cases, many of which have been unsolved for decades. Genealogists worked on these cases in collaboration with law enforcement agencies. Raw DNA data files were uploaded to the genealogy websites GEDmatch and FamilyTreeDNA, and identification was made by tracing the family trees of relatives who were predicted to be close genetic matches in the database. Such searches have far-reaching consequences because they affect not just those who have consented to upload their DNA results to these databases but also all of their relatives, regardless of whether or not they have taken a DNA test. This article provides an overview of the methods used, the potential privacy and security issues, and the wider implications for society. There is an urgent need for forensic scientists, bioethicists, law enforcement agencies, genetic genealogists and other interested parties to work together to produce international guidelines and policies to ensure that the techniques are used responsibly and effectively.


Assuntos
Crime , Impressões Digitais de DNA , Bases de Dados de Ácidos Nucleicos , Aplicação da Lei , Linhagem , Cromossomos Humanos Y , Privacidade Genética , Humanos , Consentimento Livre e Esclarecido , Repetições de Microssatélites , Sequenciamento Completo do Genoma
13.
Sud Med Ekspert ; 62(2): 22-25, 2019.
Artigo em Russo | MEDLINE | ID: mdl-31213587

RESUMO

The objective of the present study was to optimize genotyping of nuclear DNA contained in human hair. The most efficient procedures for DNA isolation and typing are described taking into consideration the hair growth phase, the epithelial tissue conditions, and the number of nuclei in the near-root ends of the hair. The recommendations for the expert interpretation of the results of the molecular-genetic investigations are proposed.


Assuntos
Núcleo Celular/genética , Impressões Digitais de DNA , DNA/isolamento & purificação , Genótipo , Cabelo/citologia , Humanos
14.
Forensic Sci Int ; 301: 174-181, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31167154

RESUMO

Allele frequency data for 22 short tandem repeat loci; D18S1364, D1S1656, D13S325, D5S2800, D9S1122, D4S2366, D3S1744, D12S391, D11S2368, D21S2055, D20S482, D8S1132, D7S3048, D2S441, D19S253, D10S1248, D17S1301, D22-GATA198B05, D16S539, D6S474, D14S1434 and D15S659 from the SureID® 23comp Human DNA Identification Kit have been determined for unrelated individuals in European, South Asian and African populations. Deviations from Hardy-Weinberg equilibrium were observed in loci D1S1656 and D19S253 in European; D18S1364, D6S474 and D14S1434 in South Asian; and D9S1122 and D8S1132 in African populations (p-value <0.05). However, after Bonferroni correction no significant deviations were observed (p-value <0.002). The most discriminating loci were D1S1656 and D12S391 for European (PD=0.977), D21S2055 for South Asian (PD=0.980), and D21S2055 and D7S3048 for African (PD=0.972) populations. The match probabilities were 1 in 6.7×1025 for European, 1 in 1.4×1026 for South Asian and 1 in 1.6×1026 for African populations. These findings established the high discriminatory capacity and robustness of the tested STR loci for forensic identification and kinship testing.


Assuntos
Grupos de Populações Continentais/genética , Impressões Digitais de DNA/instrumentação , Genética Populacional , Repetições de Microssatélites , Frequência do Gene , Humanos , Reação em Cadeia da Polimerase
15.
Forensic Sci Int ; 301: 231-239, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31181408

RESUMO

The increasing use of non-laboratory-based DNA and protein detection methods promise to provide rapid investigative intelligence and support sample prioritisation. Primarily developed for human forensic or medical applications, current systems may also show utility in the field of wildlife forensic science. However, it is currently unknown whether the requirements of the wildlife forensic community can be met by current non-laboratory based tools. Given the diverse array of stakeholders and sample types commonly encountered, it is necessary to first identify the needs of the community and then try and map their needs to current instrumentation. By using a market research style questionnaire, this study identified key requirements for a non-laboratory-based system following feedback from the wildlife forensic community. Data showed that there is strong support for field-based detection methods while highlighting concerns including contamination risks and reduced quality assurance associated with non-laboratory testing. Key species and applications were identified alongside hurdles to implementation and adoption. Broadly, the requirements align with many of the developmental drivers that have led to the rise of in-field portable detection instrumentation, specifically rapid detection within one hour, ease-of-use, and ≥95% accuracy. Several existing platforms exist that met some of the identified requirements but not all. With further collaboration between industry partners and the wildlife forensic community it is possible that new field-based systems can be developed and applied routinely.


Assuntos
Animais Selvagens/genética , Conservação dos Recursos Naturais , Impressões Digitais de DNA/instrumentação , Retroalimentação , Determinação de Necessidades de Cuidados de Saúde , Inquéritos e Questionários , Animais , Comércio , Crime , Espécies em Perigo de Extinção , Humanos , Competência Profissional , Especificidade da Espécie
16.
Forensic Sci Int ; 301: 371-381, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31212144

RESUMO

Different stakeholders use forensic DNA databases for different purposes; for example, law enforcement agencies use them as an investigative tool to identify suspects, and criminologists use them to study the offending patterns of unidentified suspects. A number of researchers have already studied their effectiveness, but none has performed an overview of the relevant literature. Such an overview could help future researchers and policymakers by evaluating their creation, use and expansion. Using a systematic review, this article synthesizes the most relevant research into the effectiveness of forensic DNA databases published between January 1985 and March 2018. We report the results of the selected studies and look deeper into the evidence by evaluating the relationship between the purpose, content, and effectiveness of DNA databases, three inseparable elements in this type of research. We classify the studies by purposes: (i) detection and clearance; (ii) deterrence; and (iii) criminological scientific knowledge. Each category uses different measurements to evaluate effectiveness. The majority of these studies report positive results, supporting the assumption that DNA databases are an effective tool for the police, society, and criminologists.


Assuntos
Direito Penal , Bases de Dados de Ácidos Nucleicos , Impressões Digitais de DNA , Humanos
17.
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
18.
J Forensic Leg Med ; 66: 4-7, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31176279

RESUMO

One of the main characteristics of the Mafia of Gargano is their use of ritual murders: they shoot their victims in the face and then conceal the corpses in the numerous natural ravines present in the Gargano area. Skeletal remains are often recovered in a poor state of preservation under particular conditions related to the environmental situation. Humidity, temperature and environmental contaminants could be considered very important for forensic examinations and are strictly related to the bone preservation status. One of the most important analyses is the identification of the victim: the success rate is linked to the condition of the bones. During military investigations in the Gargano area, several bones were recovered and analyzed in a karst ravine about 30 m deep. The forensic examination highlighted the presence of fly puparia from an intact human femur. The colonization of the inner bone cavity by a species of the Heleomyzidae family is described for the first time. Puparia, despite not being identified at the species level, are described and illustrated and their potential role in the degradation of the victim's DNA is discussed. This work increases our knowledge about the effects of Diptera in the taphonomic process underlying the need of a multidisciplinary approach to skeletal investigations.


Assuntos
Restos Mortais , Dípteros , Fêmur/patologia , Pupa , Animais , Degradação Necrótica do DNA , Impressões Digitais de DNA , Entomologia , Antropologia Forense , Humanos , Itália , Larva
19.
Comp Immunol Microbiol Infect Dis ; 64: 168-175, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31174694

RESUMO

In spite of high vaccination coverage in the Expanded Program of Immunization (EPI), pertussis has not been eradicated yet and the re-emergence of the disease is still reported worldwide. The genetic divergence study of circulating clinical strains of Bordetella pertussis among the population with high vaccination coverage is a useful tool to have an insight in the understanding of genetic patterns of this bacterium and deviation of them from vaccine strains. Different methods are accessible for studying of Bordetella pertussis that can perform appropriate assessment between populations. Strains used in this study were a collection of two pertussis vaccine strains used to create killed pertussis vaccine over years at Razi Vaccine and Serum Research Institute, 10 clinical and 2 reference strains (ATCC9797 and Tohama I) in Multilocus Sequence Typing (MLST), Pulsed-Field Gel Electrophoresis (PFGE), and serotyping. The genetic profiles of vaccine working and master seeds showed no important change(s) in frequencies of fingerprint types investigated in the vaccine strains and had homogeneity in PFGE method where the clinical isolates showed diversity in genetic profile. Serotyping method showed that all of 10 clinical strains expressing Fim 3. In MLST study, seven housekeeping genes including adk, pgm, fum C, tyr B, gly A, pep A and icd were analyzed which showed no changes in the sequence of clinical and vaccine strains with 100% homology. The genes that cause pathogenicity like ptxC, tcfA and fhaB were also evaluated and the results illustrated heterogeneity in the vaccine and circulating strains.


Assuntos
Bordetella pertussis/genética , Coqueluche/microbiologia , Técnicas de Tipagem Bacteriana , Bordetella pertussis/classificação , Impressões Digitais de DNA , Eletroforese em Gel de Campo Pulsado , Perfil Genético , Variação Genética , Genótipo , Tipagem de Sequências Multilocus , Vacina contra Coqueluche , Sorotipagem
20.
Fa Yi Xue Za Zhi ; 35(2): 210-215, 2019 Apr.
Artigo em Inglês, Chinês | MEDLINE | ID: mdl-31135117

RESUMO

Abstract: Objective To evaluate the efficiency of REPLI-g® Single Cell Kit for sample DNA amplification, and explore its application value in forensic trace DNA amplification. Methods Three DNA extraction kits were selected to extract DNA from peripheral blood of 10 unrelated individuals. The DNA yield and purity of the three DNA extraction kits were compared. According to the results of comparison, one DNA sample was selected to concentrate and dilute, then used as the initial sample of whole genome amplification (WGA). REPLI-g® Single Cell Kit was used to amplify the initial sample at the whole genome level. The amplification yield and amplification times were calculated, and the distribution of DNA fragments was detected by agarose gel electrophoresis. Goldeneye® DNA ID System 20A Kit was used to perform the STR typing of the initial sample and DNA samples amplified at the whole genome level to evaluate the performance of REPLI-g® Single Cell Kit in trace DNA amplication in terms of purity and yield as well as the success rate of STR typing. Results After comparison, one DNA sample was selected from QIAsymphony® DNA Investigator® Kit extracts to concentrate and dilute as the initial sample of WGA. After amplifying the whole genome of a series of initial samples by REPLI-g® Single Cell Kit, the lowest average of amplification yield reached 8.77×103 ng, while the average of the corresponding amplification times reached 1.40×106. DNA fragments were large and concentrated. The STR typing success rate of WGA samples became lower with the decrease of initial samples used, but when the amount of samples was lower than 0.5 ng, the STR typing success rate of samples after DNA WGA was higher than that of samples without DNA WGA. Conclusion REPLI-g® Single Cell Kit can increase the yield of template DNA. Especially for trace DNA, the STR typing success rate can be improved to a certain extent.


Assuntos
Impressões Digitais de DNA , Técnicas de Amplificação de Ácido Nucleico , Análise de Sequência de DNA/métodos , DNA , Humanos , Repetições de Microssatélites , Técnicas de Amplificação de Ácido Nucleico/métodos , Técnicas de Amplificação de Ácido Nucleico/normas
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