CTC Detection via Immunofluorescence and FISH for CRPC

The patent segments the CTC detection process into multiple independent detection channels: EpCAM-based selection for traditional CTCs, cytokeratin staining for CK- CTCs, and size-based identification for small CTCs. This segmentation allows each method to target specific CTC subpopulations without interfering with others, thereby improving overall detection coverage while maintaining the precision of each individual method.

The patent creates a universal CTC detection platform that performs multiple functions simultaneously: it detects EpCAM-positive traditional CTCs, CK-negative CTCs, and small CTCs using a single integrated system. The platform uses multi-parametric analysis combining EpCAM, cytokeratin, and size measurements to identify all CTC subpopulations in one assay, eliminating the need for separate detection methods.

The patent introduces cytokeratin staining as an intermediary marker to distinguish small CTCs from white blood cells. Since small CTCs and WBCs are similar in size, the cytokeratin stain serves as a mediating identifier that allows the detection system to differentiate between these two cell types based on protein expression rather than size alone, thereby maintaining both speed and accuracy.

The patent changes the detection parameter from size-only to a combination of size and cytokeratin expression. By adding the cytokeratin parameter to the detection criteria, the system can identify small CTCs that would otherwise be indistinguishable from WBCs based on size alone, thus improving measurement precision without sacrificing detection speed.

The patent implements feedback through CTC biomarker monitoring to guide therapy sequencing decisions. By measuring CTC counts and characteristics before and during treatment, clinicians receive real-time feedback on treatment response, allowing them to adjust the sequencing of therapy agents based on actual patient response rather than following fixed protocols, thereby simplifying the decision-making process.

The patent performs preliminary action by identifying CTC biomarker signatures before initiating therapy sequencing. By characterizing the CTC population and their molecular profiles in advance, clinicians can predict which therapy agents are most likely to be effective, allowing for optimized therapy sequencing from the outset rather than trial-and-error approaches.