Programa de Pós-graduação em Genética e Biologia Molecular
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Navegando Programa de Pós-graduação em Genética e Biologia Molecular por Por Orientador "Lacerda, Elisângela de Paula Silveira"
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Item Métodos diagnósticos para o vírus mayaro: revisão sistemática e avaliação molecular em pacientes arbovirose like em unidade municipal de sáude de Goiânia-Goiás(Universidade Federal de Goiás, 2020-02-17) Abrantes, Gabrielly Regis; Anunciaçāo, Carlos Eduardo; http://lattes.cnpq.br/4354412874919580; Lacerda, Elisângela de Paula Silveira; http://lattes.cnpq.br/9390789693192751; Lacerda, Elisângela de Paula Silveira; Anunciaçāo, Carlos Eduardo; Lino, Guilherme; Mrué, FátimaDiseases caused by viruses are considered an often neglected public health problem that affects thousands of people worldwide, every year. Symptoms like fever, arthralgia and rash. classify patients as syndromic dengue or arbovirus like. The differential diagnosis of arbovoresis is by serological and molecular techniques. The present study aimed to evaluate diagnostic techniques in studies published in the literature in order to develop a Systematic Literature Review on the detection of the Mayaro virus that allows health professionals to carry out an efficient laboratory diagnosis, as well as the application of molecular tests for the diagnosis of MAYV. The available literature was evaluated by systematic review, focusing on the diagnosis of the Mayaro virus in humans and its methodologies. The systematic review was carried out through an exploratory study in the databases: Scielo (Scientific Eletronic Library Online), Medline (Medical Literature Analysis and Retrieval System Online), Scopus database, PubMed, LILACS from June 8, 2019 to 02 November 2019, selecting scientific articles according to the criteria of the PRISMA (2009) recommendation. The data showed that the circulation of the Mayaro virus in urban regions is already well established worldwide and that the misdiagnosis is frequent due to similar symptoms with other arboviruses. There is a wide use of varied methods over time in the diagnosis for this arbovirus, requiring further research to implement a standardized, sensitive and specific method. In a second stage of the research, between May to August 2017 and January to June 2018, 452 samples of patients with syndromic dengue symptoms (arboviruses like) were screened. The samples were sent to the Molecular Genetics and Cytogenetics Laboratory of the Federal University of Goiás, where they were subjected to molecular screening by Polymerase Chain Reaction followed by Reverse Transcriptase (RT-PCR) reaction. All positive cases for the first PCR were confirmed on RT-qPCR for viral detection. Thirteen positive samples went through both tests and thus were subjected to Sanger sequencing. In this experimental study, 17 samples were positive for the Mayaro virus infection (RT-PCR and RT-qPCR), and co-infections with the Dengue virus were also found by molecular screening carried out in parallel for the Dengue virus by RT-PCR and RT-qPCR. The positive samples for MAYV / DENV were analyzed phylogenetically, suggesting circulation of Dengue serotype 2 and a Peruvian strain of Mayaro Virus (strain IQ 4235) in Goiânia.Item Análise da herança do DNA do Trypanosoma cruzi em parentais e progênies de chagásicos de famílias de São Felipe – Bahia(Universidade Federal de Goiás, 2019-02-26) Santos, Eduardo José dos; Cardoso, Clever Gomes; http://lattes.cnpq.br/9545455455623006; Lacerda, Elisângela de Paula Silveira; http://lattes.cnpq.br/9390789693192751; Teixeira, Antônio Raimundo Lima Cruz; Zapata, Marco Túlio Antônio GarciaThe Trypanosoma cruzi, etiologic agent of Chagas disease is a flagellate protozoan transmitted by insect bugs (Reduvidae:Triatomine) to humans and wild mammals. The T. cruzi contains DNA in the nucleus (nDNA) and in the mitochondrion (kDNA). The kinetoplastid protozoan comprises maxicircles and and minicircles kDNA network sequences that integrate in the host’s cell genome. Insertional integrations of kDNA sequences bear high affinity to retrotransposable LINE-1 randomly distributed in the genome of somatic and germ line cells, and, thus generate an increasing diversity due to recombination and reshuffling. However, the profiles of kDNA integrations into the human genome in different Brazilian Ecosystems are unknown. In this regard, the ratio of transfer of T. cruzi minicircles sequences into the human genome was south. The study consisted of analyses of 36 members of two families from São Felipe-Bahia where Chagas disease is endemic. The study showed that 22% (8/36) families’ members had the diagnoses of the disease by immunofluorescence (IF) and enzyme-linked immunosorbant assay (ELISA). Yet, the PCR assays showed 61% (22/36) of families’ members had kDNA and nDNA assays positive for the T. cruzi infection. In order to detect the kDNA integration sites in the human genome, a tpTAIL-PCR (Targeted Primer Thermal Assymetric Interlaced-PCR), which consisted in combination of primers derived from T. cruzi minicircle conserved sequence with human LINE-1 sequence derived primers, was employed. The amplicon sequences obtained were subjected to homology analyses in Blast software. The hybrid kDNA-human DNA sequences revealed lateral and vertical transfer of T. cruzi minicircles into the genome of Chagas parents and progeny. Unravel of dynamics of mutations contributes to define variable profiles of kDNA minicircle sequence insertions into the human genome at different Ecosystems.Item Avaliação sorológica e molecular de pacientes com quadro clínico de dengue símile atendidos no hospital das forças armadas(Universidade Federal de Goiás, 2019-02-25) Sousa, Adriano Roberto Vieira de; Lacerda, Elisângela de Paula Silveira; http://lattes.cnpq.br/9390789693192751; Lacerda, Elisângela de Paula Silveira; Zapata, Marco Túlio; Sobrinho, Carlos de Almeida BaptistaArboviruses are a public health problem due to outbreaks and diseases caused in several countries around the world. They are transmitted to humans and other animals through the bite of mosquito vectors, mainly of the genus Aedes. Dengue, Zika, Chikungunya and Mayaro viruses are arboviruses that share similar clinical symptoms (dengue-like), commonly fever, rash, myalgia and arthralgia. The differential diagnosis is performed through serology and / or molecular biology techniques. In Brazil these arboviruses are endemic and have a wide geographic distribution. Due to its reemergence, several rapid diagnostic kits are available in the market, and studies are needed to prove the effectiveness of these tests. The present study aims to evaluate the efficacy of two rapid lateral flow immunochromatography diagnostic kits - Orange Life® and Bahiafarma® . Both kits are available in the market. During the period from February to April 2017, 421 samples of whole blood were collected from dengue-like patients at the Hospital das Armed Forces, Brasília-DF. The hospital performed a serological screening for the following antigens - Dengue NS1, Dengue Anti-IgM/IgG and Chikungunya IgM using the Orange Life® kit. After sorting, two groups were selected according to inclusion and exclusion criteria based on Serological Profiles. Group 1 was 58 non-reactive samples for Dengue NS1, Dengue Anti-IgM/IgG and Chikungunya IgM antigens. Group 2 was 24 reagent samples for Dengue NS1 antigen. The samples were sent to the Laboratory of Molecular Genetics and Cytogenetics of the Federal University of Goiás where they were screened using the kit recommended by the Ministry of Health-Bahiafarma® Zika Combo Anti-IgM/IgG virus. Polymerase Chain Reaction (Real-time and Conventional) techniques were used for the detection of viral RNA. The molecular results for Group 1 were 4 positive samples for Dengue, 5 for Zika, 17 for Chikungunya and 2 for Mayaro. Two positive samples for Mayaro were sequenced, observing homology with other available sequences. For Group 2, 6 samples were positive for Dengue, 5 for Zika and 3 Chikungunya. Comparing the molecular results, the serological kit of Bahiafarma® Zika Combo Anti-IgM/IgG virus presented 100% specificity with a negative predictive value of 87.8%. Orange Life® serological kits showed - Dengue NS1 42.8% and 61.5% sensitivity and specificity; Dengue IgM 93% specificity with 83.56% negative predictive value; Chikungunya IgM 72.2% specificity with 56.4% negative predictive value. The results obtained in this work demonstrate that the tests evaluated have lower sensitivity and specificity than the recommended ones when the molecular results are compared. Health professionals are advised to choose the most appropriate test according to detection windows by watching the onset days of symptoms.