Looi L M, Yap S F, Cheah P L
Department of Pathology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
Ann Acad Med Singap. 1997 Nov;26(6):750-3.
Fresh frozen neoplastic tissues from 70 infiltrating ductal breast carcinomas were analysed for cytosolic oestrogen receptor (ER) protein content using a solid phase enzyme immunoassay (EIA) method based on a "sandwich" principle (Abbott ER-EIA monoclonal). Formalin-fixed, paraffin-embedded sections from the same carcinomas were examined for nuclear immunoreactivity against a monoclonal antibody for ER protein (Dako) using the standard avidin-biotin complex immunoperoxidase (IP) method after microwave antigen retrieval. The degree of ER positivity by IP was also scored according to a visual estimation of the percentage of cells expressing immunopositivity and the intensity of staining. Twenty-eight (40%) of the carcinomas were ER-positive by EIA and 34 (48.6%) were positive by IP. Twenty-five (35.7%) were ER-positive and 33 (47.1%) were ER-negative by both methods. Nine (12.9%) were ER-negative by EIA but were positive by IP, this discrepancy being ascribed to sampling inadequacy for EIA. However, 3 (4.3%) tumours were ER-positive by EIA and negative by IP. This discrepancy may be variously due to inadequate antigen retrieval, faulty technique and the possibility that the two methods do not measure identical ER proteins. IP appears to have an advantage over EIA in that it has a higher pick-up rate, does not require fresh tissue and can be applied to archival material. However, to reduce false negative estimations, it may be necessary to run IP staining using more than one ER antibody. Standardisation of the IP method for ER is desirable before this method is to be widely adopted in Malaysian laboratories. Quantitation of ER positivity by IP scoring correlated poorly with actual cytosolic levels. Caution should be exercised in attaching patient management value to visual IP scoring.
采用基于“三明治”原理的固相酶免疫分析(EIA)方法(雅培ER - EIA单克隆法),分析了70例浸润性导管乳腺癌的新鲜冷冻肿瘤组织中的胞质雌激素受体(ER)蛋白含量。对同一肿瘤的福尔马林固定、石蜡包埋切片,在微波抗原修复后,使用标准抗生物素蛋白 - 生物素复合物免疫过氧化物酶(IP)法,检测针对ER蛋白的单克隆抗体的核免疫反应性。IP法检测的ER阳性程度也根据对表达免疫阳性细胞百分比和染色强度的视觉估计进行评分。通过EIA检测,28例(40%)肿瘤为ER阳性;通过IP法检测,34例(48.6%)为阳性。两种方法均显示,25例(35.7%)为ER阳性,33例(47.1%)为ER阴性。9例(12.9%)通过EIA检测为ER阴性,但通过IP法检测为阳性,这种差异归因于EIA取样不足。然而,3例(4.3%)肿瘤通过EIA检测为ER阳性,通过IP法检测为阴性。这种差异可能是由于抗原修复不足、技术错误以及两种方法测量的ER蛋白可能不同等多种原因造成的。IP法似乎比EIA法更具优势,因为它的检出率更高,不需要新鲜组织,并且可以应用于存档材料。然而,为了减少假阴性估计,可能有必要使用多种ER抗体进行IP染色。在马来西亚实验室广泛采用IP法检测ER之前,需要对该方法进行标准化。通过IP评分对ER阳性进行定量与实际胞质水平的相关性较差。在将患者管理价值与视觉IP评分相关联时应谨慎。