Percorrer por autor "Mukan, Olena"
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- Genetic identification of human skeletal remains in forensic context: a reviewPublication . Cainé, Laura; Henriques, Madalena; Rohovska, Adelina; Sousa, Bárbara; Afonso Costa, Heloísa; Correia Dias, Maria Helena; Rodrigues, Joana; Franco, Magda; Mukan, Olena; Nascimento, Rui; Vânia Mofreita; Amorim, AntónioBackground/Objectives: Genetic identification of human skeletal remains plays a pivotal role in forensic investigations when other traditional or primary methods are not appropriate. Decomposition, storage and environmental conditions often leave the skeletal structure as the only basis for identification. This review synthesizes current methodologies and technological advances in damaged DNA extraction and analysis, emphasizing the forensic relevance of skeletal remains for genetic identification. Methods: A comprehensive literature analysis highlights the basis of genetic identification; sampling that considers intrinsic and extrinsic factors influencing the DNA yield and its quality; pre-treatment methods; extraction protocols that are suitable for its sensitivity; genetic marker panels that allow for human identification; and statistical evaluation and analysis of the results. The last chapter demonstrates the real-world impact of genetic identification on historical cases, underscoring its broader significance in legal, humanitarian, and socio-historical contexts, supporting a critical evaluation of best practices, methodological robustness, and ethical considerations within the field. Results: Teeth, femur and the petrous portion of temporal bone are the main samples used for genetic analysis. STR profiling and mitochondrial DNA are the gold standard markers for skeletal human identification. Minimally destructive protocols that enhance a high DNA yield are chosen, with silica-based methods being highlighted in the extraction protocols. Next-Generation Sequencing techniques have also improved analytical outcomes, by enabling high-throughput data generation, increased coverage depth, nucleotide-level sequence data, and high-level multiplexing of genetic targets. Conclusions: This review provides a comprehensive framework for researchers and practitioners seeking to optimize genetic identification workflows in forensic sciences and bioarcheology. These methodological advances have significantly increased identification success rates, especially in cases involving degraded or limited skeletal remains. Reviews such as this one help us to identify methodological gaps, ethical concerns, and future research directions, thereby establishing best practices when working with highly degraded skeletal material, supporting more reliable, standardized, and legally defensible applications of genetic identification in forensic, archeological, and humanitarian contexts.
- Hepatitis A: Ultrasensitive enzyme-linked fluorescence assay in occasional detection of prior contact in a medico-legal sample In LisbonPublication . Mofreita, Vânia; Cainé, Laura; Rodrigues, Joana; Nascimento, Rui; Mukan, Olena; Fadoni, Jennifer; Corte Real, F.; Amorim, António
- In house Real-Time PCR multiplex for simultaneous detection of human influenza A and B virus and respiratory syncytial virusPublication . Fadoni, Jennifer; Cainé, Laura; Rodrigues, Joana; Mofreita, Vânia; Nascimento, Rui; Mukan, Olena; Corte Real, F.; Amorim, António
- Next Generation Sequencing for variant detection of SARS-CoV-2 in a medico-legal context in the second year of the PandemicPublication . Rodrigues, Joana; Cainé, Laura; Mofreita, Vânia; Nascimento, Rui; Mukan, Olena; Fadoni, Jennifer; Corte Real, F.; Amorim, António
- SARS-COV-2 Variants in the region of Lisbon: Comparation and validation study between RT-PCR and NGS methodologiesPublication . Franco, Magda; Cainé, Laura; Rodrigues, Joana; Mofreita, Vânia; Nascimento, Rui; Mukan, Olena; Fadoni, Jennifer; Corte-Real, Francisco; Amorim, AntónioSARS-CoV-2 is a recent coronavirus that appeared in the end of 2019. The World Health Organization named the infection, caused by this new coronavirus, Coronavirus Disease 2019 (COVID-19). This disease was considered a pandemic on March 11th 2020 by the same organization. Coronaviruses rapidly acquire new mutations and, consequently, new variants keep emerging. There are some variants with an associated risk in the increase of the transmissibility and that cause more severe disease and reduction in the neutralization by antibodies and, for this reason, are called variants of concern. The aim of this project was to identify the main VOCs and VOIs circulating in the region of Lisbon, applying the methodology of real time RT-PCR in cadavers that tested positive for SARS-CoV-2. To meet this goal, we used three assays: Allplex™ SARS-CoV-2 Variants I, Allplex™ SARS-CoV-2 Variants II and Allplex™ SARS-CoV-2 Variants V. The first one detects defining mutations of the Alpha, Beta and Gamma variants (N501Y, E454K and HV69/70del), the second assay detects mutations present in the Delta, Beta, Gamma and California variants (L452R, K417T, K417N and W152C) and the third one detects defining mutations of the Delta and Lambda variants (L452R, P681R, L452Q and F690S). In addition, we also wanted to understand if these RT-PCR assays were efficient to correctly identify these variants, comparing the results obtained to the reference methods for the determination of variants – Next Genration Sequencing (NGS). Two different NGS methodologies were used to compare the results – NGS-ONT (Oxford Nanopore Technologies) and NGS-Sanger based. We concluded that 30% of the samples belonged to the Alpha variant and 70% belonged to the Delta variant and that, in general, the three RT-PCR assays applied in this study were efficient in correctly identifying these two variants, based on the comparison to the NGS results.