Research Projects

Many detection techniques such as PAP, nucleic acid hybridizations (southern blot, in situ hybridization, Hybrid Capture-II (gold standard), and amplification assays (PCR, RT-PCR) were in practice for the screening and diagnosis of cervical cancer. However, the confirmatory test is still the histopathology and the immunofluorescence. Both confirmatory tests are costly (INR 1400-2000), time taking (2-3 days), and require sophisticated infrastructure and skilled personnel for data interpretation. Thus, the development of rapid, non-invasive, sensitive, and cost-effective detection methods of transformed cervical cancer cells in at-risk women may have the potential to be used for confirmatory diagnosis and as an alternative to immunofluorescence/cytopathology during the treatment and management of the disease. Considering this, we propose to develop a simple, cost-effective, and visual detection system for cervical cancer by using magnetic nanoparticles (MNPs) coupled capturing and quantum dot coupled detecting nano complex for HPV-induced cervical cancer. This test will detect both E6 and E7 proteins, which is expressed only in HPV-induced cervical carcinoma using Magnetic Nanoparticles Coupled Quantum Dots Immuno-Nano-Fluorescence Assay (MNCQDINFA). We have already developed the prototype to visually detect the E7 protein of HPV-16 as a proof of concept and was already patented. We propose to elaborate this assay for a broad range, highly specific, and sensitive detection for High-risk HPV virus-induced cervical cancer. This method will produce a visual fluorescence exclusively in the presence of transformed cervical cancer cells under UV light. Briefly, polyclonal antibodies against N- and C-terminal domains of E7 and E6 protein of HPV 16 and 18 will be generated and bio-conjugated on non-fluorescent magnetic nanoparticles (for capturing complex) and on fluorescent hydrophilic quantum dots (for detection complex) after complete characterization of functionality. The cytology samples from the patient will be processed to release the total proteins in the form of clear supernatant and incubated with MNPs-Ab complex for 30 min. The MNPs bound E7/E6 oncoprotein will be separated using a magnet to remove all other non-specific proteins from the processed patient samples. QDs-Ab detecting complex will be incubated for 30 min to form MNPs-E7/E6-QDs complex. This nano-complex will be washed in the presence of a magnet to remove the unbound QDs/other impurities and again re-suspended. The fluorescence in UV light of the above solution confirms the presence of cervical cancer. This test will be highly sensitive/specific, simple, robust, rapid (less than 60 Min), cost-effective (less than 100 rupees), visual (QD fluorescence), and reproducible. Due to its simplicity, this detection system will also be used for the diagnosis of cervical cancer at primary healthcare centers situated in remote areas of India as well as other developing countries.