Detection of breast cancer micrometastases in axillary lymph nodes by using polymerase chain reaction.

摘要: Abstract Breast cancer micrometastases in axillary lymph nodes have been detected by serial sectioning and immunohistochemistry, shown to prognostic significance. We used polymerase chain reaction (PCR) see whether we could further improve the detection rate of micrometastases. Fifty-seven from patients with breast were examined histologically assess proportion tumor involvement. Immunohistochemical staining use an anti-keratin 19 antibody confirmed histological findings. Reverse transcription PCR was then performed on extracted RNA using K19 primers, all 18 involved yielded expected 460-base pair product. Of 39 negative nodes, 4 (10%) gave bands detectable ethidium a 10 (28%) after Southern hybridization. To increase sensitivity two stage amplification nested product found without cancer, as well patients. This be genuine low level expression endogenous mRNA template, not derived pseudogene. Reducing number cycles steps did allow sufficient discrimination between normal those which only The optimal “cut-off” point distinguish remained at 40 PCR, marker, has demonstrated this study cancer: is limited specificity marker.