Research Projects

Rationale: Hepatitis B virus (HBV) remains a public health issue worldwide, with over 290 million chronic carriers and significant morbidity and mortality due to liver disease. Early and accurate diagnosis is essential for clinical management and prevention of disease transmission. Conventional assays like ELISA and PCR, though helpful, are time-consuming and expensive and need central laboratory facilities—hence not very practical in poor-resource settings. Sensitive, rapid, and affordable point-of-care diagnostic platforms for HBV detection is urgently needed. Recent advances in molecular diagnostics, such as CRISPR-based detection platforms, offer highly programmable and specific alternatives to traditional assays. In combination with isothermal amplification technologies such as Recombinase Polymerase Amplification (RPA), CRISPR systems such as Cas12 and Cas9 can offer high-speed nucleic acid detection without the need for thermal cycling. For additional amplification of sensitivity and accuracy of readout of such systems, nanomaterials such as two-dimensional (2D) quantum dots (QDs) offer unique optical properties and high surface-to-volume ratios favorable to signal amplification and biomolecule conjugation. Scientific Objective: 1. 1. To synthesize and characterize 2D quantum dots. 2. To integrate 2D QDs with CRISPR-RPA output for point-of-care diagnostic platforms. 3. To validate the platform using clinical samples and compare performance with standard PCR methods. Hypothesis: We hypothesize that a CRISPR-Cas-based detection system can offer a highly sensitive, specific, and rapid diagnostic platform for HBV for point-of-care applications when paired with RPA for target amplification and 2D quantum dots for signal enhancement. Methods: 1. Design specific guide RNAs targeting conserved regions of the HBV genome. 2. Use RPA for isothermal amplification of target DNA sequences. 3. Functionalize 2D quantum dots (e.g., MoS₂ QDs, graphene QDs) with CRISPR-detection components. 4. Develop a fluorescence or colorimetric readout system triggered by target recognition and QD interaction. 5. Test the platform with synthetic and clinical HBV samples to determine limit of detection (LOD), specificity, and reproducibility. Significance: The goal is to provide a portable, low-cost diagnostic tool that can help with early HBV detection in areas with limited resources. A unique combination of cutting-edge technologies is demonstrated by the use of 2D quantum dots for sensitive readouts, RPA for quick amplification, and CRISPR for specific nucleic acid recognition. If successful, this platform could be expanded to include other viral infections and revolutionise the way that infectious disease monitoring is currently done, particularly in low- and middle-income nations. On a fundamental level, it will also advance knowledge of interfaces between biological systems and nanomaterials and their function in the development of next-generation biosensors.