Biological sensors are widely applied in agriculture, biomedicine, food, healthcare, environmental monitoring, water quality, forensics, drug development, etc.. Particularly the utilization of microfluidic technology has become prevalent in the development and manufacturing of biosensors for miniaturization, automation, and integration. Microfluidic biosensors have distinct advantages, including enhanced diffusive timescales, controlled concentration gradients, high throughput, high precision fluid manipulation, stable reaction environments and high sensitivity. From the perspective of sensor fabrication, bonding remains the crucial pathway in the pursuit of integrating microfluidic technology with biosensor chips, while various bonding methods are employed across different application domains. This paper delves into the classification, progress, and challenges associated with these bonding methods corresponding with various microfluidic biosensors in diverse applications. The review presented herein highlights the latest advancements in microfluidic biosensors based on diverse bonding methods, underscoring their significant application prospects and developmental potential within these fields.