Approaches To Designing Immobilisation Sequences To Improve Preconcentration Assay Of Prolactin-Inducible Protein (PIP) mRNA Using Complementary Molecular Beacon

Abstract

Breast cancer is a current prevalent disease found mainly in women. The cancer when reaches metastases is expensive and difficult to treat. Diagnosis of breast cancer is usually done by mammography and prognosis has headed to the direction of detecting certain biomarkers in the blood or serum done by RT-PCR. Studies have shown that prolactin-inducible protein (PIP), human epidermal growth factor receptor 2 (HER2) and cytokeratin-19 (CK19) are three among the biomarkers prognostic for breast cancer. Therefore, quantification of these markers could provide important information about breast cancer metastasis. Previous studies have proposed a preconcentration method to improve the limit of detection of these biomarkers in buffer and serum samples using molecular beacon (MB) and designed immobilization sequences. In this report, different approaches in designing immobilization oligonucleotides and their effect on the preconcentration assay were explored. Results showed that the design of immobilization oligonucleotides did not have a significant effect on the preconcentration factor whether the MB-washed eluent was collected hot or cooled.