手工免疫组化检测非小细胞肺癌ALK蛋白表达的对比分析

Comparative study of ALK antibody with manual and automatic immunohistochemical detection in non-small cell lung cancer

目的探讨手工免疫组化(immunohistochemistry,IHC)法检测不同克隆号ALK抗体在间变性淋巴瘤激酶(anaplastie lymphoma kinase,ALK)融合的非小细胞肺癌(non-small cell lung cancer,NSCLC)中ALK蛋白表达,并与全自动免疫组化染色法进行比较。方法选取60例NSCLC石蜡样本,运用4种抗体D5F3(Ventana)、D5F3(Cell Signaling)、1A4/1H7(Ori Gene)、5A4(Abcam)联合常规手工IHC分别检测NSCLC中ALK蛋白水平,与抗体D5F3(Ventana)采用全自动IHC(Bench Mark)结果进行对比分析。结果 D5F3(Ventana+Bench Mark)检测发现ALK融合阳性NSCLC者32例,ALK融合阴性NSCLC者28例,经FISH法验证结果一致。采用手工IHC法检测4种抗体D5F3(Ventana)、D5F3(Cell Signaling)、1A4/1H7、5A4染色灵敏性分别为93.8%、84.4%、93.8%、56.3%,特异性均为100%,与D5F3(Ventana+Bench Mark)染色结果的一致性分别为96.7%、91.7%、96.7%、76.7%。手术切除大标本的ALK表达优于活检小标本。结论手工IHC法简便经济、易于普及,选择高灵敏性、特异性的抗体,可用于中小医院临床筛查ALK融合NSCLC,染色阳性病例再行抗体D5F3(Ventana)全自动IHC或FISH法检测。

Purpose To explore the immunohistochemical （IHC） expression of ALK antibodies with different clones in anaplastic lym- phoma kinase （ALK） gene fusion non-small cell lung cancer （NSCLC）. Methods ALK expression in 60 NSCLCs were detected by IHC including autostainer （DSF3, Ventana ＋ BenchMark） and manual staining using 4 different antibodies of DSF3 （Ventana） , DSF3 （Cell Signaling） , 1A4/1H7 （OriGene）, 5A4 （Abeam） , and all cases were verified with ALK FISH. Their expressions with 4 anti- bodies were compared with those by D5F3 （Ventana ＋ BenchMark）. Results 32 ALK gene rearrangement NSCLCs and 28 negative cases were identified by FISH and DSF3 （Ventana ＋ BenchMark）. The sensitivity of DSF3 （Ventana） , DSF3 （Cell Signaling） , 1A4/ 1H7 （OriGene）, 5A4 （Abcam） was 93.8%, 84.4%, 93.8%, 56.3%, and all the speciticity was 100%. The consistency with D5F3 （Ventana ＋ BenchMark） was 96. 7% , 91.7% , 96. 7% and 76.7% , respectively. The validity of immunohistochemical staining in surgical resection specimens was better than in small biopsies. Conclusion Effective routine manual immunohistochemistry with high-affinity antibody clone may provide a more economic and widespread pre-screening technique.