Seed health testing is a cornerstone of global food security, yet traditional diagnostic workflows often struggle to balance speed, sensitivity, and regulatory confidence under low-prevalence and heterogeneous seed lot conditions. This review synthesizes recent advances in molecular diagnostics (PCR, qPCR, LAMP, and digital PCR), non-destructive imaging technologies (hyperspectral, X-ray, and thermal imaging), and data-driven analytical approaches for pathogen detection in seeds. Emphasis is placed on the practical integration of these tools within high-throughput, ISO/IEC 17025-compliant laboratory workflows, highlighting their respective strengths, limitations, and roles in risk-based decision-making. Comparative discussions address cost, sensitivity, turnaround time, and field deployability across diagnostic platforms, supported by crop- and pathogen-specific examples. Emerging approaches such as CRISPR-based biosensing, advanced sequencing, and imaging-assisted analytics are discussed in the context of validation, regulatory acceptance, and operational feasibility. By focusing on implementation rather than conceptual frameworks, this review provides a pragmatic reference for laboratories, regulators, and seed companies seeking to modernize seed health testing while safeguarding trade integrity and biosecurity.

Seed health testing is undergoing a rapid transformation as emerging technologies supplement and, in some cases, replace conventional diagnostic methods. This review synthesizes recent advances in molecular diagnostics (PCR, qPCR, LAMP, and metabarcoding), non-destructive imaging approaches (hyperspectral, multispectral and X-ray) and AI-assisted pattern recognition for pathogen detection in seeds. Emphasis is placed on integrating these tools into high-throughput seed quality programs, with case studies from vegetable, ornamental and field crop systems. We highlight current limitations in cost, regulatory alignment and global standardization, while identifying future opportunities for rapid, sensitive and field-deployable testing. This review aims to guide researchers, seed technologists and policymakers toward more efficient and reliable seed health assurance strategies.