Gold-facilitated in situ hybridization: a bright-field autometallographic alternative to fluorescence in situ hybridization for detection of Her-2/neu gene amplification.
Gold-facilitated in situ hybridization: a bright-field autometallographic alternative to fluorescence in situ hybridization for detection of Her-2/neu gene amplification.
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金促进原位杂交：荧光原位杂交的明场自金相替代方法，用于检测 Her-2/neu 基因扩增。
DOI：
10.1016/s0002-9440(10)61106-6
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发表时间：
2002
期刊：
影响因子：
--
通讯作者：
Hainfeld,James
中科院分区：
文献类型：
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作者：
Tubbs,Raymond;Pettay,James;Skacel,Marek;Powell,Richard;Stoler,Mark;Roche,Patrick;Hainfeld,James
荧光原位杂交（FISH）是一种很好的方法，用于分析基因扩增原位，但相关的组织形态是困难的，因为暗场可视化。研究了用于评估HER-2/neu基因扩增的明视野测定法的验证。链霉亲和素-纳米金被用于使用GoldEnhance基于金的自显影、催化的报告沉积和生物素标记的探针来产生明场基因拷贝信号。100例浸润性乳腺癌进行了评估，其中FISH基因拷贝结果，mRNA和癌蛋白基因表达，是已知的。自显影信号可在不使用油浸显微镜的情况下进行定性评价。结果与间接和直接标记FISH相关性良好。自显影金基原位杂交是一种很有前途的明场检测HER-2/neu基因扩增的评估。
Fluorescence in situ hybridization (FISH) represents an excellent method for profiling gene amplification in situ, but correlation with tissue morphology is difficult because of dark-field visualization. Validation of a bright-field assay for assessment of HER-2/neu gene amplification was investigated. Streptavidin-Nanogold was used to generate bright-field gene copy signals using GoldEnhance gold-based autometallography, catalyzed reported deposition, and a biotin-labeled probe. One hundred cases of invasive breast carcinoma were evaluated for which FISH gene copy results, and mRNA and oncoprotein gene expression, were known. Autometallographic signals were qualitatively evaluable without the use of oil immersion microscopy. Results correlated well with indirect and direct label FISH. Autometallographic gold-based in situ hybridization represents a promising bright-field assay for the assessment of HER-2/neu gene amplification.