Dual-colour chromogenic in situ hybridization for testing of HER-2 oncogene amplification in archival breast tumours.

摘要: Chromogenic in situ hybridization (CISH), which uses an enzymatic reaction to detect the hybridized DNA probe, is a new alternative fluorescence (FISH) for assessment of HER-2 oncogene amplification status breast cancer. The main advantage CISH over FISH use bright-field microscopy, rapid and allows histopathological evaluation tumour tissue sections. disadvantage has been single thereby making it necessary hybridize control probe (chromosome 17 centromere) on adjacent section. present paper presents efficient protocol dual-colour (dc-CISH) based co-hybridization probes chromosome centromere. were detected sequentially with antibodies digoxigenin biotin secondary antibody polymers labelled horseradish peroxidase alkaline phosphatase. was visualized tetramethyl benzidine (green product) phosphatase New Fuchsin (red product). accuracy method verified by comparing results four cell lines 40 samples those obtained using (Vysis Inc.). dc-CISH showed high concordance (91%, Kappa coefficient = 0.82). It concluded that CISH, enables copy number ratio (HER-2/17 conjunction proper ease microscopy.