A differential PCR assay for the detection of c-erbB 2 amplification used in a prospective study of breast cancer.
OBJECTIVE
To establish a robust differential polymerase chain reaction (PCR) assay for the detection of c-erbB 2 amplification in breast cancer that can be used in a routine pathology laboratory. Once established, the assay was used in a prospective study of breast tumours to investigate the relation between c-erbB 2 amplification and both recognised prognostic features and short term clinical outcome.


METHODS
The differential 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 electrophoresed on a highly resolving agarose gel.


RESULTS
The differential PCR assay was shown to be accurate and reproducible using the conditions 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 recurrence during the follow up period and 73% of these patients had amplification of c-erbB 2.


CONCLUSIONS
A significant association was found between c-erbB 2 amplification and early disease recurrence. This assay could be used to provide a marker for poor prognosis in breast cancer.
Jennings
B A
BA
Molecular Genetics Department, Norfolk and Norwich Hospital, UK.
Hadfield
J E
JE
Worsley
S D
SD
Girling
A
A
Willis
G
G
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001943:Breast Neoplasms / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D005260:Female; D005500:Follow-Up Studies; D018734:Genes, erbB-2; D006801:Humans; D008207:Lymphatic Metastasis; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D011379:Prognosis; D011446:Prospective Studies