Detection of preconcentrated mRNA through a molecular beacon assay for the early detection of breast cancer metastasis
Ong, Amy M.
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Among all cancers, breast cancer has the second highest incidence rate among women. However, early detection of the disease has led to increased treatment options for patients. To obtain accurate diagnoses, biomarkers for breast cancer can be monitored and the accuracy is even further increased when two or more biomarkers are monitored. To detect multiple biomarkers in a sample, molecular beacons (MB) can be used due to their high specificity and sensitivity. Two molecular beacons were designed specific to mRNA transcripts of two different breast cancer biomarkers for simultaneous detection in a single sample. Each MB uses a different fluorophore-quencher pair; therefore the fluorescence output caused by the target-mRNA-MB interaction can be monitored for each biomarker simultaneously. The total time for the designed assay is significantly shorter than current methods based on RT-PCR. The limit of detection for each biomarker has been reduced with pre-concentration using beads and a capture oligonucleotide sequence complementary to the monitored biomarkers.
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