In Malaysia, about 1,740 new cases of cervical cancer are diagnosed every year and 991 of them die annually because the cancer cells that are spreading unconsciously in the body are not detected in time. Thus, rapid and simple detection methods for prevention and identification are required to solve the morbidity and mortality related to the pathogenic virus. This research has focused on developing a simple, rapid, and novel multi-detection method to detect HPV 16, 18, and 58 at significantly lower concentrations, using these strains as reference to establish effective rapid detection strategies. Firstly, the surface morphology is analyzed using scanning electron microscopy (SEM) and 3D profiler to determine the width and spacing of the finger electrode to 50 μm and 1.5 μm, respectively. Subsequently, the physiochemical properties of colloidal gold Nanoparticles (AuNPs) were investigated using transmission electron microscopy (TEM) (D: ±9.5 nm) and ultraviolet (UV-Vis) (521 nm). Nanoscale IDEs were developed to determine and reduce the sensor size. In addition, IDEs were functionalized with colloidal AuNPs and constructed with APTES as linkers. The optimized IDEs were then used to detect HPV ssDNA by the presented two-step mechanism after surface modification by APTES. APTES links the modified HPV-designed ssDNA probe with the carboxyl group (-COOH) through immobilization by covalent binding via amine (-NH2). Consequently, each HPV ssDNA probe was used to detect its specific ssDNA target during hybridization. The detection principle works by detecting changes in the electrical current of the IDE, bridging the source and drain junction to detect immobilization of the HPV sDNA probe and hybridization with the target sDNA. During the hybridization process, the measured current of the target was higher than its probe as drastic changes in charges generated a spike in the current profile. It was also found that the sensor showed sensitivity for HPV ssDNA target in a linear range with concentrations from 1 aM to 100 μM. The regression coefficient for HPV 16, 18 and 58 was 0.99857, 0.98928 and 0.99583 respectively, and the sensitivity test for HPV 16, 18 and 58 measured at 0.00302 AM-1, 0.074 AM-1 and 0.28089 AM-1 respectively. With this analysis, the limit of detection (LOD) was approximately 0.1 aM, 0.02 aM, and 0.025 aM for HPV 16, 18, and 58 respectively. Next, HPV 18 and 58 were detected in the real sample with an accuracy of 90.42% and 91.92% also validated with PCR significantly. The reusability of the biosensors for HPV 16, HPV 18, and HPV 58 is up to two weeks after their development. Therefore, the first IDEs electrode biosensor based on Au-deposited NPs for early detection of cervical cancer is presented in this research thesis.
