中国口腔颌面外科杂志

中国口腔颌面外科杂志 ›› 2019, Vol. 17 ›› Issue (3): 216-220.doi: 10.19438/j.cjoms.2019.03.005

• 论著 • 上一篇    下一篇

免疫组织化学双染技术在唾液腺淋巴上皮性病变中的应用评价

顾挺，胡宇华，夏荣辉，田臻，王丽珍，张春叶*，李江*   

  1. 上海交通大学医学院附属第九人民医院 口腔医学院 口腔病理科，国家口腔疾病临床医学研究中心，上海市口腔医学重点实验室，上海市口腔医学研究所，上海 200011
  • 收稿日期:2018-10-17 修回日期:2019-01-16 出版日期:2019-05-20 发布日期:2019-06-21
  • 通讯作者: 张春叶，E-mail：yezi1806@126.com；李江，E-mail:lijiang182000@126.com。
  • 作者简介:顾挺（1981-），男，学士，主管技师，E-mail：yuemachun@126.com

Application of double stained immunohistochemistry technology in lymphoepithelial lesions of salivary gland

GU-Ting, HU Yu-hua, XIA Rong-hui, TIAN Zhen, WANG Li-zhen, ZHANG Chun-ye, LI Jiang   

  1. Department of Oral Pathology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine;National Clinical Research Center for Oral Disease;Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology.Shanghai 200011, China
  • Received:2018-10-17 Revised:2019-01-16 Online:2019-05-20 Published:2019-06-21

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摘要： 目的: 根据抗体阳性表达的不同部位（细胞核、细胞质和细胞膜）、染色顺序及匹配不同的显色系统，探讨免疫组织化学双染技术在唾液腺淋巴上皮性病变诊断中的最佳染色方法。方法: 挑选良性淋巴上皮病（benign lymphoepithelial lesion，BLEL）、淋巴上皮癌（lymphoepithelial carcinoma，LEC）和黏膜相关淋巴组织结外边缘区B细胞淋巴瘤（extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue，MALT淋巴瘤）各20例，分别进行AE1/AE3、Ki-67、CD20cy抗体的免疫组织化学单染和两两组合的双染检测。每个病例依据不同抗体的表达部位（细胞核、细胞质及细胞膜）、不同的抗体染色顺序和显色剂，分别采用3种双染方法检测，即①先Ki-67（DAB显色），再AE1/AE3或CD20cy（AEC显色）；② 先AE1/AE3或CD20cy（DAB显色），再Ki-67（AEC显色）；③ 先Ki-67（AEC显色），再AE1/AE3或CD20cy（BCIP/NBT显色）。所得结果均与单染相比较，采用SPSS 17.0软件包对数据进行χ2检验和配对t检验，分析染色强度和阳性比率有无差异。结果: 所有双染方法中抗体定位均准确，但方法1中各抗体染色强度（P=0.765）和阳性比率（P>0.05）均无显著差异，而方法2和方法3中抗体的阳性比率和染色强度均有不同程度下降（P<0.05）。结论: 免疫组织化学双染技术在唾液腺淋巴上皮性病变中的最佳染色方法为先进行阳性定位于细胞核如Ki-67的染色，配合使用DAB显色剂，后进行阳性定位于细胞膜或细胞质如AE1/AE3或CD20cy的染色，配合使用AEC显色剂。

关键词: 免疫组织化学双染技术, 淋巴上皮病, 淋巴上皮癌, MALT淋巴瘤, 阳性强度, 阳性率

Abstract: PURPOSE: Depending on the sites of positive antibody expression (nucleus, cytoplasm and membrane), this study was aimed to investigate the best method of applying double stained immunohistochemistry technology in lymphoepithelial lesions of salivary gland. METHODS: The expression of AE1/AE3，Ki-67 and CD20cy was examined by single stained immunohistochemistry and double stained immunohistochemistry respectively in 20 benign lymphoepithelial lesions (BLEL), 20 lymphoepithelial carcinomas (LEC) and 20 extranodal marginal zone lymphomas of mucosa-associated lymphoid tissue (MALT) . Depending on the different sites of antibody expression (nucleus, cytoplasm and membrane), the sequence of staining and chromogenic system, three methods of double stained immunohistochemistry were performed: ① Ki-67(DAB chromogen)，AE1/AE3 or CD20cy(AEC chromogen); ②AE1/AE3 or CD20cy(DAB chromogen)，Ki-67(AEC chromogen); ③ Ki-67(AEC chromogen)，AE1/AE3 or CD20cy(BCIP/NBT chromogen). The results of double stained immunohistochemistry were compared with single stained immunohistochemistry. The data were analyzed by using SPSS 17.0 software package. RESULTS: The results of immunohistochemistry demonstrated specificity and accuracy of the immunostaining procedures in all three methods, no significant difference of the positive strength (P=0.765) and positive ratio of antibody (P>0.05) was found in method 1. Otherwise, there were significant differences of the positive strength and positive ratio of antibody in method 2 and 3 (P<0.05). CONCLUSIONS: The best method is that nucleus positive antibody should be chosen first (Ki-67), followed by DAB chromogen, cytoplasm and membrane positive antibody should be chosen second(AE1/AE3 or CD20cy), combined with AEC chromogen for double stained immunohistochemistry in lymphoepithelial lesions of salivary gland.

Key words: Double stained immunohistochemistry technology, Benign lymphoepithelial lesion, Lymphoepithelial carcinoma, Extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue, Strength of positivity, Positive ratio

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