A DIFFERENTIAL PCR ASSAY FOR THE DETECTION OF C-ERBB-2 AMPLIFICATION USED IN A PROSPECTIVE-STUDY OF BREAST-CANCER

Aims T-establish a robust differential polymerase chain reaction (PCR) assay for the detection of c-erbB 2 amplification in breast cancer th at can be used in a routine pathology laboratory Once established, the assay was used in a prospective study of breast tumours to investigat e the relation between c-erbB 2 amplification and both recognised prog nostic features and short term clinical outcome. Methods-The different ial PCR was used for the co-amplification of c-erbB 2 and a reference gene from 48 tumour DNA samples and control DNA samples. The ratio of the two genes was determined by image analysis of the PCR products ele ctrophoresed on a highly resolving agarose gel. Results-The differenti al PCR assay was shown to be accurate and reproducible using the condi tions outlined. Twenty six per cent of the breast cancer patients were shown to have c-erbB 2 amplification in their tumour biopsies. Twenty eight per cent of the patients died of their disease or had disease r ecurrence during the follow up period and 73% of these patients had am plification of c-erbB 2. Conclusions-A significant association was fou nd between c-erbB 2 amplification and early disease recurrence. This a ssay could be used to provide a marker for poor prognosis in breast ca ncer.