Accurate and early detection of microbial pathogens plays a vital role in disease control, food safety, and environmental monitoring. This chapter provides an overview of traditional and modern sensing techniques for detecting microbial pathogens, which discusses the principles and applications of these methods and their developments and trends. The introduction underlines the importance and significance of pathogen sensing in public health and industry. Then, the traditional approaches, including microscopy and staining techniques and, in particular, Gram staining and fluorescence microscopy, are represented to view and distinguish between microorganisms. Immunological approaches that exploit antigen–antibody reactions will be shown as a reliable means to identify pathogens. Then, molecular methods, including polymerase chain reaction and its derivatives, isothermal amplification-based techniques, and next-generation sequencing (NGS), are represented based on their superior sensitivity to detect microbial DNA or RNA. The chapter addresses biosensor technologies, electrochemical, optical, and nanotechnology-based biosensors. Such detection technologies provide rapid and portable means to sense pathogens based on sensor designs and breakthrough materials technology. Emerging technologies such as CRISPR-based detection systems, AI and machine learning applications, microfluidic devices, and wearable sensors, among others, are transformative tools in the next-generation pathogen sensing of the future. Other issues include detection limits, cost, and integration into real-world applications, all within the context of providing perspectives on future trends. In conclusion, the importance of integrating advanced technologies with classical methods to improve the efficiency and reliability of pathogen detection is stressed. This book chapter will be highly valued by researchers and practitioners of microbiology, diagnostics, and biosensing.