Evaluation of ER, HER2 and SPATA19 Genes Expression in Clinical Samples of Breast Cancer Using NASBA-ELISA

The most important issue in diagnosing human breast cancer is the ability to detect the early stages of cancer, which can help the healing process. Therefore, new diagnostic methods for breast cancer focus on molecular approaches. The aim of this work is to develop the NABA-ELISA method for the diagnosis of breast cancer in clinical samples, which is used for the first time to evaluate HER2, ER and SPATA19 biomarkers simultaneously. The possibility of expressing the SPATA19 gene, a specific biomarker for testis cancer, was also investigated in breast cancer. Specific primers and biotinylated probes were designed separately for each of the target genes. RT-PCR and NASBA reactions were done on total RNA extracted from the clinical tumor tissues. The DIG-labeled NASBA product was detected by the ELISA method using the biotinylated probe and anti-DIG antibody-enzyme conjugate. The ELISA reaction showed obvious color change and significant absorption in the positive samples. Of 15 samples tested; 11, 4 and 3 samples were positive for ER, HER2 and SPATA19 genes, respectively. Comparison of real-time RT-PCR with NASBA-ELISA showed the same results for both ER and HER2 genes. There was no significant relationship in the expression of the SPATA19 gene with ER and HER2 genes in these specimens. Also, no significant relationship was observed between SPATA19 gene expression and breast cancer in these samples. In this study, we developed a simple, fast, and reliable NASBA-ELISA method that can detect cases of breast cancer.