Comparação do índice de ki-67 obtido através de contagem manual e de análise automatizada ImmunoRatio em amostras de tumores mamários caninos, carcinomas mamários felinos e mastocitomas
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Data
2019
Autores
Serras, Maria João Cardoso
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Resumo
O Ki-67 trata-se de uma proteína nuclear expressa em todas as fases do ciclo celular,
excepto na fase de repouso, utilizada como marcador de prognóstico em vários tumores.
A contagem manual é um método amplamente utilizado para avaliar o índice de Ki-
67, no entanto é moroso e pouco prático. Com este estudo, pretendeu-se comparar a contagem
manual com a análise automatizada (ImmunoRatio) no cálculo do índice de Ki-67, definir a
melhor metodologia para a análise de amostras através do plug-in ImmunoRatio e ainda
verificar a capacidade do ImmunoRatio na análise de imagens de menor ampliação (50x).
Foram analisadas um total de 18 amostras (2 mastocitomas, 11 carcinomas mamários
felinos e 5 tumores mamários caninos). Em amostras homogéneas foram selecionados cinco
campos aleatórios e, em amostras heterogéneas, três campos aleatórios e três campos com um
elevado número de células Ki-67-positivas (hotspots).
Verificou-se uma correlação forte, positiva e estatisticamente significativa e uma
concordância quase perfeita entre a contagem manual e as diferentes análises realizadas no
plug-in ImmunoRatio (ImmunoRatio 200x sem estroma, ImmunoRatio 400x sem estroma e
ImmunoRatio 200x com estroma). O estudo sugere ainda que a utilização do plug-in em
imagens de menor ampliação (50x) permite obter resultados idênticos à média das contagens
manuais da mesma amostra.
Pode assim concluir-se que a utilização do ImmunoRatio pode ser uma alternativa
viável e rápida à contagem manual. No entanto, sugere-se que nos casos em que se obtenham
valores próximos dos valores limite indicativos de agressividade, se reconfirme com a
contagem manual.
Ki-67 is a nuclear protein expressed in every phase of the cell cycle, except in the resting phase, which is used as a prognostic marker for various types of tumours. Manual counting is a widely used method to evaluate the Ki-67 index, even though it is a time-consuming and laborious. In this study, the objectives were to compare the method of manual counting with automated analysis (ImmunoRatio) when determining the Ki-67 index, to define the best methodology for sample analysis through the ImmunoRatio plug-in, and verify its capacity to analyse lower magnification images (50x). A total of 18 samples were analysed (2 mast cell tumours, 11 feline mammary carcinomas and 5 canine mammary tumours). In homogeneous samples, five random fields were chosen and, in heterogeneous samples, three random fields and three fields with a high number of Ki-67 positive cells (hotspots) were selected. A strong correlation, positive and statistically significant, and an almost perfect concordance between the manual count and the different analyses performed with the ImmunoRatio plug-in (ImmunoRatio 200x without stroma, ImmunoRatio 400x with stroma and ImmunoRatio 200x with stroma) were verified. This study also suggests that using the plug-in with lower magnification images (50x) allows for identical results to the manual count average for the same sample. It can thus be concluded that the use of ImmunoRatio can be a viable and faster alternative to manual counting. Nevertheless, it is suggested that, in the cases where approximate values to the aggressiveness indicative limit are obtained, manual counting should be used to confirm these values.
Ki-67 is a nuclear protein expressed in every phase of the cell cycle, except in the resting phase, which is used as a prognostic marker for various types of tumours. Manual counting is a widely used method to evaluate the Ki-67 index, even though it is a time-consuming and laborious. In this study, the objectives were to compare the method of manual counting with automated analysis (ImmunoRatio) when determining the Ki-67 index, to define the best methodology for sample analysis through the ImmunoRatio plug-in, and verify its capacity to analyse lower magnification images (50x). A total of 18 samples were analysed (2 mast cell tumours, 11 feline mammary carcinomas and 5 canine mammary tumours). In homogeneous samples, five random fields were chosen and, in heterogeneous samples, three random fields and three fields with a high number of Ki-67 positive cells (hotspots) were selected. A strong correlation, positive and statistically significant, and an almost perfect concordance between the manual count and the different analyses performed with the ImmunoRatio plug-in (ImmunoRatio 200x without stroma, ImmunoRatio 400x with stroma and ImmunoRatio 200x with stroma) were verified. This study also suggests that using the plug-in with lower magnification images (50x) allows for identical results to the manual count average for the same sample. It can thus be concluded that the use of ImmunoRatio can be a viable and faster alternative to manual counting. Nevertheless, it is suggested that, in the cases where approximate values to the aggressiveness indicative limit are obtained, manual counting should be used to confirm these values.
Descrição
Orientação: Pedro Faísca
Palavras-chave
MESTRADO INTEGRADO EM MEDICINA VETERINÁRIA, VETERINÁRIA, MEDICINA VETERINÁRIA, VETERINARY MEDICINE, CANÍDEOS, CANIDS, FELÍDEOS, FELIDS, CÃES, DOGS, GATOS, CATS, CARCINOMAS, GLÂNDULAS MAMÁRIAS, NEOPLASIAS, IMUNOHISTOQUÍMICA, MASTOCITOMAS, CARCINOMAS, MAMMARY GLANDS, NEOPLASMS, IMMUNOHISTOCHEMISTRY, MAST CELL TUMOR, IMMUNORATIO, KI-67