The ascorbic acid (AA) is a biomarker that can be used to detect the symptoms of severe disorders such as scurvy, Parkinson’s, Alzheimer’s, and cardiovascular diseases. In this work, a simple and effective sensor model is developed to diagnose the presence of AA samples. To develop the sensor, a tapered single-mode optical fiber has been used with the well-known phenomenon of localized surface plasmon resonance (LSPR). For LSPR, the tapered region is immobilized with synthesized gold nanoparticles (AuNPs) and zinc oxide nanoparticles (ZnO-NPs) whose absorbance peak wavelengths appear at 519 nm and 370 nm, respectively. On the basis of nanoparticles (NPs) configurations, two different biosensor probes are developed. In the first one, the sensing region is immobilized with AuNPs and named Probe I. In the second probe, the immobilized layer of AuNPs is further coated with a layer of ZnO-NPs, and a resultant probe is termed as Probe II. The characterizations of synthesized AuNPs and developed fiber probes are done by the ultraviolet-visible (UV-vis) spectrophotometer, high-resolution transmission electron microscope (HR-TEM), atomic force microscopy (AFM), and scanning electron microscope (SEM). To enhance the selectivity, a sensing region of probes is functionalized with ascorbate oxidase enzyme that oxidizes the AA in the presence of oxygen. The response of developed sensor probes is authenticated by sensing the samples of AA in the range from 500 nM to 1 mM, which covers the range of AA found in human bodies, i.e., 40 μM – 120 μM. The performance analysis of the developed sensor probes has been done in terms of their stability, reproducibility, reusability, and selectivity. To observe the stability of AA, a pH-test has also been done that results in a better solubility of AA molecules in phosphate-buffered saline (PBS) solution.