Yin Li, Wang Wan, Liu Fan, Zhang Zhenjiang, Tesfagaber Weldu, Liu Renqiang, Li Fang, Bu Zhigao, Zhu Yuanmao, Zhao Dongming
State Key Laboratory for Animal Disease Control and Prevention, National African Swine Fever Para-reference Laboratory, National High Containment Facilities for Animal Diseases Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, PR China.
State Key Laboratory for Animal Disease Control and Prevention, National African Swine Fever Para-reference Laboratory, National High Containment Facilities for Animal Diseases Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, PR China.
J Virol Methods. 2025 Sep;337:115173. doi: 10.1016/j.jviromet.2025.115173. Epub 2025 May 3.
African swine fever (ASF) is a lethal infectious disease affecting domestic and wild pigs, caused by the African swine fever virus (ASFV), with a mortality rate of up to 100 %. The evolving prevalence and variation of ASFV has led to the emergence of low-virulence strains, which induced chronic infections and posed challenges in nucleic acid-based diagnostics due to potential false negatives. This underscores the urgent need for reliable antibody monitoring to facilitate early diagnosis. In this study, based on a highly attenuated BK2258 cell-adapted strain HLJ18/BK33, we established an immunoperoxidase monolayer assay (IPMA) for ASFV antibodies detection. After optimization using a total of 608 pig sera, the performance of the assay was better than that of the commercial iELISA with higher sensitivity and specificity. The newly established IPMA method demonstrated high specificity with no cross-reactivity with positive sera for six other important porcine pathogens. The IPMA method developed in the present study could serve as the potential gold standard for serological diagnosis and evaluation of other detection methods for ASFV antibodies, owing to its high sensitivity and specificity. Furthermore, the IPMA method will provide a new and effective strategy for ASF monitoring, prevention and control in China.
非洲猪瘟（ASF）是一种由非洲猪瘟病毒（ASFV）引起的、影响家猪和野猪的致命性传染病，死亡率高达100%。ASFV不断变化的流行情况和变异导致了低毒力毒株的出现，这些毒株引发慢性感染，并因可能出现假阴性结果而给基于核酸的诊断带来挑战。这凸显了对可靠抗体监测的迫切需求，以促进早期诊断。在本研究中，基于高度减毒的适应BK2258细胞株HLJ18/BK33，我们建立了一种用于检测ASFV抗体的免疫过氧化物酶单层试验（IPMA）。在使用总共608份猪血清进行优化后，该试验的性能优于商业间接ELISA，具有更高的灵敏度和特异性。新建立的IPMA方法显示出高特异性，与其他六种重要猪病原体的阳性血清无交叉反应。本研究中开发的IPMA方法因其高灵敏度和特异性，可作为ASFV抗体血清学诊断和其他检测方法评估的潜在金标准。此外，IPMA方法将为中国的ASF监测、预防和控制提供一种新的有效策略。
