Life Sciences & Biotechnology
Title : | Detection of Viral RNA by Two-Fold Cyclic Amplification Method |
Area of research : | Life Sciences & Biotechnology |
Focus area : | Molecular Biology and Diagnostic Technologies |
Principal Investigator : | Dr. Ankan Dutta Chowdhury, AMITY University, Kolkata, West Bengal (700153) |
Contact info : | dc_ankan@yahoo.co.in |
Details
Executive Summary : | The project aims to develop a double amplification cyclic process for detecting RNAs, using a two-layer ssDNA and ssRNA nanocomposite system. The process can amplify RNAs to large numbers, potentially replacing RT-PCR or other virus detection protocols and offering cost-effective, rapid, and easy handling.
The study focuses on a cyclic amplification process using ssDNA probes as reverse complements to target RNA sequence segments. The fluorophore, Rhodamine Rh, is labeled at the 5' end and a thiol group at the other end for further functionalization. Uniform magnetic Fe3O4 nanoparticles MNP with carboxylic functionalization can be modified for thiol reactive group by melamide addition. The MNP quenches the fluorescence of the Rh in the MNP-probe-Rh complex, which can bind to the viral RNA to form DNA/RNA heteroduplexes. In the presence of DSN enzyme, it can specifically cleave the ssDNA from the heteroduplex, releasing the fluorophore Rh from the MNP and recovering fluorescence. The RNA strand is free to bind to another MNP-probe-Rh and release more fluorophores. The fluorescence increases up to a detectable level for an indication of the existence of the target RNA. |
Total Budget (INR): | 11,22,000 |
Organizations involved