Blood transfusion safety constitutes a cornerstone of clinical healthcare, with infectious disease biomarker detection being pivotal for risk prevention. Conventional testing methods suffer from limitations including low efficiency and inadequate multi-target simultaneous screening. Leveraging its unique capability for simultaneous amplification across multiple targets in a single system, multiplex PCR technology has emerged as an efficient solution. This study examines the technical characteristics and clinical requirements of this approach, detailing practical implementation strategies such as multi-target system design, sample pretreatment protocols, optimized reaction conditions, result interpretation, and quality control standards. Through standardized design and operational procedures, the technology enhances both processing speed and accuracy, providing robust support for clinical blood transfusion safety and demonstrating significant clinical application value.