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Advisor(s)
Abstract(s)
Introdução – A preservação a longo prazo de lâminas de citologia aspirativa por agulha fina (CAAF) é um requisito essencial nos laboratórios de citopatologia para a posterior realização de imunocitoquímica (ICQ). A ICQ contribui para um diagnóstico correto e completo, sendo essencial a preservação morfológica e antigénica a longo prazo para obter resultados confiáveis. Neste estudo pretende-se avaliar a imunoexpressão dos antigénios TTF1, p40 e cromogranina A em amostras de CAAF do pulmão retiradas do arquivo e coradas pelos métodos de: i) Papanicolaou (Pap); ii) May-Grünwald Giemsa (MGG); iii) preservadas em polietilenoglicol (PEG); e iv) processadas como citobloco (CB). Métodos – Foram selecionados do arquivo 24 exames de CAAF com diagnóstico de carcinoma primário do pulmão, com amostra processada por cada um dos protocolos em estudo (Pap, MGG, PEG e CB). Com base no diagnóstico foi realizada imunomarcação com anticorpos primários anti-TTF1 (adenocarcinomas), antí-p40 (carcinomas pavimentocelulares) e anti-cromogranina A (carcinomas neuroendócrinos). A qualidade da imunomarcação foi aferida por dois avaliadores independentes com recurso a uma escala, com classificação entre 0 e 27 pontos, e que compreende os parâmetros: preservação morfológica, intensidade da marcação específica, sensibilidade, especificidade e contraste. Resultados – A pontuação média obtida para os métodos Pap, MGG, PEG e CB foi de 21,58 (±4,54), 11,79 (±1,88), 22,25 (±5,30), 26,31 (±1,21) pontos, respetivamente. O CB conseguiu resultados superiores aos restantes protocolos em estudo (p<0,05). Quando comparados os protocolos a par (post-hoc de Tuckey), os únicos que não apresentaram diferenças estatisticamente significativas entre si foram Pap e PEG (p=0,814). Conclusões – O CB é o protocolo de eleição para a realização de ICQ nas amostras e para os antigénios em estudo. Os métodos Pap e PEG apresentaram perda de imunormarcação, podendo levar a resultados falso-negativos. O protocolo de MGG não obteve imunomarcação em nenhuma amostra.
ABSTRACT: Background – Long-term preservation of fine-needle aspiration cytology slides is an essential requirement in cytopathology laboratories for the eventual performance of immunocytochemistry. ICQ contributes to a correct and complete diagnosis, considering that long-term morphological and antigenic preservation is essential to obtain reliable results. In this study, we intend to evaluate and compare the immunoexpression of TTF1, p40, and chromogranin A antigens in lung samples taken from the archive and stained with: i) Papanicolaou (Pap); ii) May-Grünwald Giemsa (MGG); iii) preserved in polyethylene glycol (PEG); and iv) processed as cell-block. Methods – Twenty-four fine needle aspiration cytology samples diagnosed as primary lung carcinoma with a sample processed by each of the protocols studied (Pap, MGG, PEG, and CB) were selected from the archive. Based on the diagnosis, immunostaining was performed with primary antibodies anti-TTF1 (adenocarcinomas), anti-p40 (squamous cell carcinomas), and anti-chromogranin A (neuroendocrine carcinomas). The quality of immunostaining was evaluated by two independent observers using an evaluation grid (rated from 0 to 27 points) that comprises parameters as: morphological preservation, specific staining intensity, sensitivity, specificity, and contrast. Results – The mean values obtained for CB, PEG, Pap, and MGG protocols were 21.58 (±4.54), 11.79 (±1.88), 22.25 (±5.30), 26.31 (±1.21) points respectively. CB achieved better results when compared to other protocols under study (p<0.05). When compared in pairs (Tuckey post-hoc) the only protocols that did not show statistically significant differences were Pap and PEG (p=0.0814). Conclusions – Cellblock is the elected protocol to perform ICQ for the samples and antigens under study. The Pap and PEG protocols showed a loss of immunostaining, which could lead to false-negative results. Immunostaining was not observed in any sample with the MGG protocol.
ABSTRACT: Background – Long-term preservation of fine-needle aspiration cytology slides is an essential requirement in cytopathology laboratories for the eventual performance of immunocytochemistry. ICQ contributes to a correct and complete diagnosis, considering that long-term morphological and antigenic preservation is essential to obtain reliable results. In this study, we intend to evaluate and compare the immunoexpression of TTF1, p40, and chromogranin A antigens in lung samples taken from the archive and stained with: i) Papanicolaou (Pap); ii) May-Grünwald Giemsa (MGG); iii) preserved in polyethylene glycol (PEG); and iv) processed as cell-block. Methods – Twenty-four fine needle aspiration cytology samples diagnosed as primary lung carcinoma with a sample processed by each of the protocols studied (Pap, MGG, PEG, and CB) were selected from the archive. Based on the diagnosis, immunostaining was performed with primary antibodies anti-TTF1 (adenocarcinomas), anti-p40 (squamous cell carcinomas), and anti-chromogranin A (neuroendocrine carcinomas). The quality of immunostaining was evaluated by two independent observers using an evaluation grid (rated from 0 to 27 points) that comprises parameters as: morphological preservation, specific staining intensity, sensitivity, specificity, and contrast. Results – The mean values obtained for CB, PEG, Pap, and MGG protocols were 21.58 (±4.54), 11.79 (±1.88), 22.25 (±5.30), 26.31 (±1.21) points respectively. CB achieved better results when compared to other protocols under study (p<0.05). When compared in pairs (Tuckey post-hoc) the only protocols that did not show statistically significant differences were Pap and PEG (p=0.0814). Conclusions – Cellblock is the elected protocol to perform ICQ for the samples and antigens under study. The Pap and PEG protocols showed a loss of immunostaining, which could lead to false-negative results. Immunostaining was not observed in any sample with the MGG protocol.
Description
Keywords
Imunocitoquímica May-Grϋnwald Giemsa Polietilenoglicol Papanicolaou Processamento Immunocytochemistry Polyethyleneglycol Sample processing
Citation
Santos B, Roque R, Pereira T, Mendonça P, André S. Imunocitoquímica em citologia aspirativa do pulmão: comparação de quatro protocolos. Saúde & Tecnologia. 2022;(26):27-35.
Publisher
Instituto Politécnico de Lisboa, Escola Superior de Tecnologia da Saúde de Lisboa