A importância da genotipagem molecular no estudo de mola hidatiforme
Data
2021-06-21
Autores
Laviola, Gabriela Marini 
Orientadores
Malinverni, Andréa Cristina de Moraes
Tipo
Dissertação de mestrado
Título da Revista
ISSN da Revista
Título de Volume
Resumo
Objetivo: Analisar a aplicabilidade da técnica de STR nas amostras parafinadas de MH, do arquivo do Departamento de Patologia. Métodos: Foram selecionadas 38 amostras de tecido placentário e as áreas de interesse (decídua e vilo corial) foram demarcadas para extração do DNA separadamente. Após a extração, 17 amostras foram incluídas no estudo, devido à concentração e qualidade do DNA. A metodologia aplicada foi a genotipagem molecular por repetições curtas em tandem – STR, utilizando Investigator IDplex Plus – Qiagen, incluindo 15 marcadores autossômicos e a Amelogenina X/Y. Além disso, as análises dos eletroferogramas foram realizadas pelo software Gene Mapper ID-X. Resultados: De acordo com o exame anatomopatológico e a revisão do patologista, dos 17 casos estudados, seis foram diagnosticados como MHC (cinco amplificaram), três como MHP (dois amplificaram), cinco conflitantes entre ANM/MHP (dois amplificaram) e três com suspeita de MHC/Mosaico (todos amplificaram). Além disso, ao realizar a comparação com a análise da STR, em cinco casos concluiu-se a necessidade do tecido materno para realizar a genotipagem (casos 1, 8, 9, 12 e 15), em oito a STR confirmou o anatomopatológico (casos 2, 3, 4, 5, 7, 10, 15 e 16), sendo dois destes com a presença do tecido materno para análise da genotipagem (casos 10 e 16), quatro foram inconclusivos (casos 6, 11, 13 e 17, amplificação de poucos marcadores) e em um caso não houve amplificação para nenhum marcador (caso 14). Conclusão: Acreditamos que a genotipagem por STR possui papel relevante na determinação da origem parental das molas hidatiformes. Em suma, apesar dos desafios e limitações encontradas na qualidade das amostras para aplicar essa metodologia, ela mostrou-se importante tanto para refinar o diagnóstico das molas hidatiformes, quanto distinguir com precisão molas hidatiformes de abortos não molares em conjunto com análise cuidadosa do anatomopatológico.
Objective: The purpose of this study is to analyze the applicability of the STR technique in paraffin-embedded molar tissues from the archive of Pathology Department. Methods: 38 samples of placental tissue were selected and the areas of interest (deciduous and chorionic villus) were demarcated for DNA extraction from each region separately. After extraction, 17 samples were included from the study due to the concentration and quality of the DNA. The applied methodology was molecular genotyping by short tandem repeats – STR using Investigator IDplex Plus – Qiagen, including 15 autosomal markers and Amelogenin X / Y. In addition, the electropherogram analyzes were performed using the Gene Mapper ID-X software. Results: According to the anatomopathological examination and the review by the pathologist, of the 17 studied cases, six were diagnosed as MHC (five amplified), three as MHP (two amplified), five conflicting between ANM/ MHP (two amplified) and three with suspicion MHC/Mosaic (all amplified). In addition, when comparing with the STR analysis, in five cases the need for maternal tissue to perform genotyping (cases 1, 8, 9, 12 and 15) was concluded, eight STR confirmed the anatomopathological (cases 2, 3, 4, 5, 7, 10, 15 and 16), two of which with the presence of maternal tissue for genotyping analysis (cases 10 and 16), four were inconclusive (cases 6, 11, 13 and 17, amplification of few markers) and in one case there was no amplification for any marker (case 14). Conclusion: We believe that molecular genotyping by STR methodology has a relevant role in determining the parental origin of hydatidiform moles. In short, despite the challenges and limitations found in the quality of the samples to apply this methodology, it proved to be important both to refine the diagnosis of hydatidiform mole and to accurately distinguish hydatidiform mole from non-molar abortions in conjunction with careful anatomopathological analysis.
Objective: The purpose of this study is to analyze the applicability of the STR technique in paraffin-embedded molar tissues from the archive of Pathology Department. Methods: 38 samples of placental tissue were selected and the areas of interest (deciduous and chorionic villus) were demarcated for DNA extraction from each region separately. After extraction, 17 samples were included from the study due to the concentration and quality of the DNA. The applied methodology was molecular genotyping by short tandem repeats – STR using Investigator IDplex Plus – Qiagen, including 15 autosomal markers and Amelogenin X / Y. In addition, the electropherogram analyzes were performed using the Gene Mapper ID-X software. Results: According to the anatomopathological examination and the review by the pathologist, of the 17 studied cases, six were diagnosed as MHC (five amplified), three as MHP (two amplified), five conflicting between ANM/ MHP (two amplified) and three with suspicion MHC/Mosaic (all amplified). In addition, when comparing with the STR analysis, in five cases the need for maternal tissue to perform genotyping (cases 1, 8, 9, 12 and 15) was concluded, eight STR confirmed the anatomopathological (cases 2, 3, 4, 5, 7, 10, 15 and 16), two of which with the presence of maternal tissue for genotyping analysis (cases 10 and 16), four were inconclusive (cases 6, 11, 13 and 17, amplification of few markers) and in one case there was no amplification for any marker (case 14). Conclusion: We believe that molecular genotyping by STR methodology has a relevant role in determining the parental origin of hydatidiform moles. In short, despite the challenges and limitations found in the quality of the samples to apply this methodology, it proved to be important both to refine the diagnosis of hydatidiform mole and to accurately distinguish hydatidiform mole from non-molar abortions in conjunction with careful anatomopathological analysis.
Descrição
Citação
LAVIOLA, Gabriela Marini. A importância da genotipagem molecular no estudo de mola hidatiforme. 2021. 113 f. Dissertação (Mestrado em Patologia) - Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP). São Paulo, 2021.