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Authors
Advisor(s)
Abstract(s)
Introdução: A citopatologia é uma área anatomopatológica utilizada para o diagnóstico de lesões
benignas e malignas. A avaliação das amostras baseia-se nas características morfológicas celulares,
coradas com colorações de rotina. A coloração de May-Grünwald Giemsa (MGG) é amplamente
utilizada, através da coloração rápida para avaliação imediata das amostras para adequação de exames
complementares a realizar. A análise morfológica das células nem sempre é suficiente para um
diagnóstico definitivo, pelo que é necessária a aplicação de técnicas complementares como a
imunocitoquímica (ICQ). Por vezes, as lâminas coradas com MGG podem ser únicas para a aplicação
de ICQ. A aplicação desta técnica em amostras previamente coradas com MGG é descrita na literatura
com resultados variáveis.
Objetivos: O presente estudo tem como objetivo a otimização e validação de protocolos ICQ em
amostras citológicas coradas com MGG, para pesquisa dos antigénios citoqueratina AE1/AE3, antigénio
leucocitário comum 45 (CD45) e fator de transcrição da tiroide (TTF-1). Também nos propomos avaliar
a qualidade da técnica ICQ e calcular a sensibilidade e especificidade de cada soro.
Materiais e métodos: Utilizámos 30 amostras, 10 para cada soro e realizámos 3 protocolos: dois
protocolos-padrão (citobloco e polietilenoglicol – PEG) e MGG. Posteriormente, efetuámos ICQ e para
todas as lâminas foi avaliada a qualidade da técnica ICQ.
Resultados: Os protocolos otimizados para os soros CD45 e citoqueratina AE1/AE3 foram validados
com valores de sensibilidade e especificidade de 100%. Por oposição, o protocolo relativo ao soro TTF1 não foi validado, devido à sensibilidade ter sido de 40%. Não foram encontradas diferenças
estatisticamente significativas entre os protocolos citobloco e PEG para nenhum dos soros.
Discussão e conclusão: A prévia coloração por MGG não parece afetar a qualidade dos resultados ICQ
para os soros CD45 e citoqueratina AE1/AE3, contrariamente ao soro TTF-1. Não encontrámos falsos
positivos, tendo sido estudada a positividade de todos os anticorpos estudados para diagnóstico
definitivo.
Introduction: Cytopathology is an anatomopathological area of diagnosis, used to diagnose benign or malignant lesions. The evaluation of the samples is based on the morphological characteristics of the cells, stained with routine stains. May-Grünwald Giemsa (MGG) stain is widely used through rapid staining to evaluate samples to adequate complementary exams to realize. However, morphological analysis is not always sufficient for a complete diagnosis, so it is necessary to apply complementary techniques such as immunocytochemistry (ICC). In some cases, the MGG stained samples are unique to apply the ICQ technique. The application of this technique to samples previously stained with MGG is reported in the literature as possible, but with inconsistent results. Aims: The aim of this study is to optimize and validate the ICC protocols in cytological samples previously stained with MGG, to test for the antigens cytokeratin AE1/AE3, leukocyte common antigen 45 (CD45) and thyroid transcription factor (TTF-1). The quality of the ICC technique was also assessed, as was the calculation of the sensitivity and specificity of each serum. Materials and methods: We used 30 samples, 10 were used for each serum and 3 protocols were carried out: standard protocols (cellblock and polyethylene glycol - PEG) and MGG. ICC was subsequently applied, and the quality was evaluated for all the slides. Results: The optimized protocols for CD45 and cytokeratin AE1/AE3 serums were validated (sensitivity and specificity = 100%). In contrast, the optimized protocol for TTF-1 serum was not validated (sensitivity = 40%). It was not found statistical significant differences between cellblock and PEG protocols for every serums. Discussion and conclusion: It can be inferred that previous staining with MGG does not significantly affect the CD45 and cytokeratin AE1/AE3 immunocytochemical results. Although negatively affects the quality of TTF-1 immunocytochemical results. False positive cases were not found, and the positivity of studied antibodies was validated with definitive diagnosis.
Introduction: Cytopathology is an anatomopathological area of diagnosis, used to diagnose benign or malignant lesions. The evaluation of the samples is based on the morphological characteristics of the cells, stained with routine stains. May-Grünwald Giemsa (MGG) stain is widely used through rapid staining to evaluate samples to adequate complementary exams to realize. However, morphological analysis is not always sufficient for a complete diagnosis, so it is necessary to apply complementary techniques such as immunocytochemistry (ICC). In some cases, the MGG stained samples are unique to apply the ICQ technique. The application of this technique to samples previously stained with MGG is reported in the literature as possible, but with inconsistent results. Aims: The aim of this study is to optimize and validate the ICC protocols in cytological samples previously stained with MGG, to test for the antigens cytokeratin AE1/AE3, leukocyte common antigen 45 (CD45) and thyroid transcription factor (TTF-1). The quality of the ICC technique was also assessed, as was the calculation of the sensitivity and specificity of each serum. Materials and methods: We used 30 samples, 10 were used for each serum and 3 protocols were carried out: standard protocols (cellblock and polyethylene glycol - PEG) and MGG. ICC was subsequently applied, and the quality was evaluated for all the slides. Results: The optimized protocols for CD45 and cytokeratin AE1/AE3 serums were validated (sensitivity and specificity = 100%). In contrast, the optimized protocol for TTF-1 serum was not validated (sensitivity = 40%). It was not found statistical significant differences between cellblock and PEG protocols for every serums. Discussion and conclusion: It can be inferred that previous staining with MGG does not significantly affect the CD45 and cytokeratin AE1/AE3 immunocytochemical results. Although negatively affects the quality of TTF-1 immunocytochemical results. False positive cases were not found, and the positivity of studied antibodies was validated with definitive diagnosis.
Description
Tese de Mestrado, Biologia Humana e Ambiente , 2023, Universidade de Lisboa, Faculdade de Ciências
Keywords
Citopatologia May-Grünwald Giemsa Imunocitoquímica Teses de mestrado - 2023