Abstract: Infectious pancreatic necrosis (IPN), classified as a Category three notifiable aquatic animal disease by the Ministry of Agriculture and Rural Affairs of China, poses a major threat to global salmonid aquaculture, particularly affecting juvenile rainbow trout (Oncorhynchus mykiss). Due to the different strains of the virus, the mortality rate of the affected fish can range from 10% to 90%. The pathogen infectious pancreatic necrosis virus (IPNV) has different molecular characteristics and significant differences in virulence among different genogroups, and genogroup 1 and 5 pose the greatest threat to rainbow trout aquaculture in China. This study aimed to establish an integrated diagnostic approach for concurrent detection and genotyping of IPNV.A probe-based real-time reverse transcription quantitative PCR (RT-qPCR) assay was designed using conserved regions of the IPNV VP2 gene to enable high-efficiency viral detection. Additionally, genogroup-specific PCR primer sets were developed targeting unique sequences of IPNV genogroups 1 and 5 for genotyping. Assay performance was systematically optimized and validated through sensitivity, specificity, and reproducibility testing. The results demonstrated that the RT-qPCR detection method established in this study exhibited high specificity and did not amplify the nucleic acid of viral haemorrhagic septicaemia virus (VHSV) or infectious haematopoietic necrosis virus (IHNV). The sensitivity was high, with detection limits as low as 10 copies/μL for both IPNV genogroups 1 and 5. The stability was excellent, with intra- and inter-batch coefficients of variation being less than 1%. Using the RT-qPCR identification and PCR genotyping method established in this study, 30 samples from different rainbow trout aquaculture regions were tested for IPNV, achieving a 100% consistency rate. The research findings indicate that the method established in this study can detect IPNV specifically, stably, and sensitively, and accurately distinguish between genogroups 1 and 5 virus strains, achieving efficient integration of pathogen identification and genotyping. This research provides effective techniques for the monitoring of IPNV in China and scientific guidance for the classified prevention and control of IPN.