This project present the design and development on the hardware and software of navigation control system for a Vision-Based Autonomous Guided Vehicle (V-AGV) by detecting and recognizing line tracking with Universal Serial Bus (USB) camera. In this project, a mobile robot, a laptop computer and a low-cost USB camera are used as the main components of the system. The USB camera will allow the system to obtain digital images directly without using frame grabbers. The vision-based navigation system structure is composed of several processes; these processes consist of Guideline Detection, Sign Detection, Obstacle Detection and Avoidance and finally Speed control. During the navigation process, the V-AGV would be able to recognize the straight and crossing track lines, detect any obstacle that appears in the navigational path, detect the sign as guidance, avoiding obstacle and perform speed control based on the current situations. In order to demonstrate the results of this proposed navigation system, a mobile root platform as been developed. The results obtained from several experiments have shown that the USB camera has performed very well in executing the proposed algorithm.