PCR approach

An alternative to immunocytochemical assays for the detection of DTC are molecular detection procedures. The nucleic acid from a sample can be amplified by PCR, so that tumor cells can be detected with high sensitivity in a heterogeneous population of cells. However, the tumor cells must have changes in its DNA or mRNA expression pattern that distinguish them from the surrounding hematopoetic cells. At the DNA level, breast carcinomas are genetically heterogeneous, with no universally applicable DNA marker available. Therefore, research has focused on RNA markers. A multimarker approach with a panel of tumor-specific mRNA markers may improve the sensitivity for the detection of DTC over single marker assays (21, 22). Many transcripts have been evaluated as "tumor-specific" markers like CK18, CK19, CK20, Mucin-1, and carcinoembryonic antigen (23-25). However, many of these transcripts can also be identified by RT-PCR in normal BM, blood, and lymph node tissue (26-28). Preanalytical depletion of the disturbing normal cell fraction (e.g., granulocytes that express CK20) or quantitative RT-PCR determinations could solve this problem.

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