Hwang Hyun Jin, Choi Yun Seong, Song Kyungyoung, Frant Maciej, Kim Jeong Hee
R&D Center, Ahram Biosystems Inc., Seoul, South Korea.
Department of Swine Diseases, National Veterinary Research Institute, Puławy, Poland.
Front Vet Sci. 2023 Jan 4;9:1037728. doi: 10.3389/fvets.2022.1037728. eCollection 2022.
African swine fever virus (ASFV) is a double-stranded DNA virus that causes African swine fever (ASF), a lethal hemorrhagic fever that is highly contagious among domestic pigs and wild boars. Due to the high mortality rates and highly contagious nature of the ASF, it is important to develop a fast detection method for ASFV with high sensitivity and specificity to take an immediate action to stop wide spread of the virulent disease. Therefore, a fast and quantitative molecular detection method of ASFV is presented in this study. A total of 24 genotypes of ASFV have been identified based on nucleic acid sequences of the major capsid protein p72. The primers and probe of the present assay was designed to detect all of the p72-based genotypes of ASFV. The turnaround time for PCR detection was within 50 min which is at least about two-times faster compared to other PCR assays. Limit of detection (LoD) was 6.91 genomic copies/reaction for the most virulent genotype II. LoD values for other genotypes were within 10-20 copies/reaction. Cross-reactivity of the assay was validated using a panel of pathogens related to swine disease, and no cross-reactivity was observed. Positive and negative clinical samples (50 samples each) obtained from sick and healthy animals, were used to validate the assay. The results showed that 100% agreement for both positive and negative samples. In summary, the assay described in this study offers the advantage of rapid detection of all genotypes of ASFV with high sensitivity and specificity. The assay is a valuable tool both in clinical and laboratory uses for sensitive and fast detection of ASFV.
非洲猪瘟病毒（ASFV）是一种双链DNA病毒，可引发非洲猪瘟（ASF），这是一种致命的出血热，在家猪和野猪中具有高度传染性。由于ASF的高死亡率和高传染性，开发一种具有高灵敏度和特异性的ASFV快速检测方法对于立即采取行动阻止这种烈性疾病的广泛传播至关重要。因此，本研究提出了一种ASFV的快速定量分子检测方法。基于主要衣壳蛋白p72的核酸序列，已鉴定出总共24种ASFV基因型。本检测方法的引物和探针旨在检测所有基于p72的ASFV基因型。PCR检测的周转时间在50分钟内，与其他PCR检测方法相比至少快约两倍。对于毒性最强的II型基因型，检测限（LoD）为6.91个基因组拷贝/反应。其他基因型的LoD值在10 - 20个拷贝/反应范围内。使用一组与猪病相关的病原体验证了该检测方法的交叉反应性，未观察到交叉反应。从患病和健康动物获得的阳性和阴性临床样本（各50份）用于验证该检测方法。结果显示，阳性和阴性样本的一致性均为100%。总之，本研究中描述的检测方法具有快速检测所有ASFV基因型的优点，具有高灵敏度和特异性。该检测方法在临床和实验室中都是用于灵敏快速检测ASFV的宝贵工具。
