Page 234 - 《水产学报》2026年第2期

2 期 刘威彤，等：传染性胰脏坏死病毒 (IPNV) 检测及区分 1、5 基因型方法的建立及应用 50 卷

Establishment and application of a method to detect and distinguish
genogroups 1 and 5 of infectious pancreatic necrosis virus (IPNV)

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LIU Weitong 1,2,3 , ZHANG Zhen , ZHAO Jingzhuang ,
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SHAO Yizhi , LU Tongyan , XU Liming 2,3*
(1. National Experimental Teaching Demonstration Center of Aquatic Science,
Shanghai Ocean University, Shanghai 201306, China;
2. Heilongjiang Fisheries Research Institute, Chinese Academy of Fishery Sciences, Harbin 150070, China;
3. Key Laboratory of Aquatic Animal Diseases and Immune Technology of Heilongjiang Province, Harbin 150070, China)

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). Mortality rates in affected fish vary from 10% to 90%, depending on the viral strain. The
pathogen infectious pancreatic necrosis virus (IPNV) has different molecular characteristics and significant differences in vir-
ulence 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 the 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. The assay's performance was systematically optimized and validated by
evaluating its sensitivity, specificity, and reproducibility. The results demonstrated that the RT-qPCR detection method estab-
lished 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 cop-
ies/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 differ-
ent 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.
Key words: coldwater fish; infectious pancreatic necrosis virus (IPNV); genotyping; quantitative real-time PCR (RT-qPCR);
rapid detection
Corresponding author: XU Liming. E-mail: xuliming@hrfri.ac.cn

Funding projects: Central Public Welfare Institutions Basic Research Funds Special Fund Project (2023TD45)

中国水产学会主办 sponsored by China Society of Fisheries https://www.china-fishery.cn
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