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Doctoral Thesis
DOI
https://doi.org/10.11606/T.5.2021.tde-10112021-111233
Document
Author
Full name
Daniela de Oliveira Francisco
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2021
Supervisor
Committee
Rave, Cintia Fridman (President)
Iwamura, Edna Sadayo Miazato
Dias, Alexandre Torchio
Jorge, Alexander Augusto de Lima
Title in Portuguese
Estudo de DNA de toque em situações simuladas para finalidades forenses
Keywords in Portuguese
DNA de toque
Extração de DNA
Genética forense
Identificação Humana
Perfil genético
Repetições de microssatélites
Abstract in Portuguese
Impressões digitais são vestígios encontrados em cenas de crimes que podem ser usados em análises forenses para a obtenção de perfis STR a partir do DNA recuperado das mesmas, técnica conhecida como DNA de Toque. Vários fatores podem afetar a obtenção e a qualidade desses perfis, como o tipo de superfície em que o DNA é depositado. No Brasil, os crimes mais recorrentes são aqueles praticados com armas brancas, os roubos de veículos e os cibercrimes, sendo que em muitos casos não se considera o uso de perfis de DNA nas investigações por se considerar que o DNA de Toque não é uma boa fonte de material biológico. O objetivo desse estudo foi analisar a viabilidade de obtenção de perfis de DNA em diferentes materiais de empunhaduras de facas, maçanetas e volantes de carros, e em teclados de computadores por meio de situações simuladas de transferência direta. Ao todo, 104 experimentos foram conduzidos a fim de avaliar a eficácia de dois kits de extração de DNA: DNA IQ System e Casework Direct Kit (ambos Promega Corporation). Nove indivíduos participaram do estudo: quatro homens e cinco mulheres. O DNA foi coletado pelo método duplo swab. As amostras foram quantificadas com o kit Quantifiler Trio (Thermo Fisher) a amplificadas com o PowerPlex® Fusion System (Promega). Foi possível obter 83 perfis STR (32 extraídas com DNA IQ e 51 com Casework), sendo que amostras extraídas com Casework Direct Kit obtiveram uma média de alelos coincidentes com as referências maior do que as com DNA IQ. Dos 83 perfis gerados, 31 (37,3%) seriam úteis para identificação do criminoso (24 de Casework 77,4%), considerando como parâmetro de utilidade do perfil, a presença de 8 de 13 marcadores STR do CODIS original. Ao considerar 8 de 20 marcadores do CODIS atual, 42 perfis (50,6%) seriam úteis (30 de Casework 71,4%). Drop-out e drop-in foram observados, em número maior para amostras extraídas com DNA IQ. Foram observadas variações no status shedder dos mesmos participantes em diferentes experimentos, dificultando a classificação em bons ou maus doadores. Experimentos com duas mulheres foram melhores para identificação do doador secundário do que ensaios com dois homens, sugerindo que mulheres podem atuar mais como removedoras de células do que depositoras. O teste Qui-quadrado mostrou que o Casework Direct Kit foi melhor para a produção de perfis úteis para uma investigação em todas as superfícies (p = 0.001). O teste Kruskal-Wallis mostrou que o tipo de superfície afeta signficativamente a recuperação de DNA para ambos os métodos de extração (p = 0.005 para DNA IQ e p = 0.002 para Casework), sendo que os volantes obtiveram os melhores resultados e as empunhaduras de facas os piores. Conclui-se que o Casework Direct Kit foi mais eficaz para análise de amostras de DNA de toque para os três tipos de materiais estudados, ainda que outros fatores interfiram na geração do perfil
Title in English
Touch DNA study in simulated situations for forensic purposes
Keywords in English
DNA extraction
Forensic genetics
Genetic profile
Human Identification
Microsatellite repeats
Touch DNA
Abstract in English
Fingerprints are traces commonly found in crime scenes that can be used in forensic analysis to obtain STR profiles from the DNA recovered from them, this technique being known as Touch DNA. Several factors can affect the obtaining and quality of these profiles, such as the type of surface on which the genetic material is deposited. In Brazil, the most recurrent crimes are those committed with bladed weapons, theft of vehicles and cyber crimes, and in many cases the use of DNA profiles in investigations is not used because touch DNA is not considered as a good source of biological material. The objective of this study was to analyze the feasibility of obtaining DNA profiles in different materials for knife handles, knobs and steering wheels for cars, and on computer keyboards through simulated situations of direct transfer. In all, 104 experiments were conducted in order to evaluate the effectiveness of two DNA extraction kits: DNA IQ System and Casework Direct Kit (both Promega Corporation). Nine individuals participated in the study: four men and five women. DNA was collected using the double swab method. The samples were quantified with the Quantifiler Trio kit (Thermo Fisher) and amplified with the PowerPlex® Fusion System (Promega). It was possible to obtain 83 STR profiles (32 extracted with DNA IQ and 51 with Casework), and samples extracted with Casework Direct Kit obtained an average of alleles coinciding with the references samples higher than those with DNA IQ. Of the 83 profiles generated, 31 of them (37.3%) would be useful for identifying the criminal (24 from Casework - 77.4%), considering as the profile utility parameter, the presence of 8 of 13 STR markers from the original CODIS. When considering 8 out of 20 current CODIS markers, 42 profiles (50.6%) would be useful (30 from Casework - 71.4%). Drop-out and drop-in were observed, although in greater numbers for samples extracted with DNA IQ. Variations in the shedder status of the same participants were observed in different experiments, making it difficult to classify as good or bad donors. Experiments with two women were better at identifying the secondary donor than trials with two men, suggesting that women may act more as cell removers than depositors. The Chi-square test showed that the Casework Direct Kit was better for producing useful profiles for an investigation on all tested surfaces (p = 0.001). The Kruskal-Wallis test showed that the type of surface significantly affects DNA recovery for both extraction methods (p = 0.005 for DNA IQ and p = 0.002 for Casework), with the steering wheels obtaining the best results and the knife handles the worst. It is concluded that the Casework Direct Kit was more effective for analyzing touch DNA samples for the three types of materials studied, although other factors, such as the type of surface or environmental quality, interfere in the generation of the profile
 
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Publishing Date
2021-11-10
 
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