Establishment of a Dual-Antigen Indirect ELISA Based on p30 and pB602L to Detect Antibodies against African Swine Fever Virus.
Establishment of a Dual-Antigen Indirect ELISA Based on p30 and pB602L to Detect Antibodies against African Swine Fever Virus.
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DOI：
10.3390/v15091845
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发表时间：
2023-08-30
期刊：
影响因子：
--
通讯作者：
Zhang G
中科院分区：
文献类型：
--
作者：
Zhou L;Song J;Wang M;Sun Z;Sun J;Tian P;Zhuang G;Zhang A;Wu Y;Zhang G
非洲猪瘟（ASF）是由非洲猪瘟病毒（ASFV）引起的一种急性、强毒性和高度致命的传染病。目前还没有有效的疫苗和诊断方法来预防和控制该病，这就突出了早期发现非洲猪瘟的重要性。在本研究中，我们选择了一种早期抗原和一种晚表达抗原共同检测靶抗体，这不仅有助于早期发现，而且提高了准确性和灵敏度。CP204L和B602L在大肠杆菌载体体系中成功表达为可溶性蛋白。通过对各种条件的优化，建立了检测ASFV抗体的双抗原间接ELISA方法。该检测与猪繁殖与呼吸综合征病毒、猪瘟病毒、猪圆环病毒2型和伪狂犬病毒抗体无交叉反应。检测asfv阳性血清的最大血清稀释度为1:1600。批内变异再现系数<5%，批间变异再现系数<10%。与市售试剂盒相比，双抗原间接ELISA具有良好的检测性能。综上所述，我们建立了一种成本低、生产流程简化、仪器数量少的检测方法。为非洲猪瘟的血清学诊断提供了一种新的方法。
African swine fever (ASF) is an acute, virulent, and highly fatal infectious disease caused by the African swine fever virus (ASFV). There is no effective vaccine or diagnostic method to prevent and control this disease currently, which highlights the significance of ASF early detection. In this study, we chose an early antigen and a late-expressed antigen to co-detect the target antibody, which not only helps in early detection but also improves accuracy and sensitivity. CP204L and B602L were successfully expressed as soluble proteins in an Escherichia coli vector system. By optimizing various conditions, a dual-antigen indirect ELISA for ASFV antibodies was established. The assay was non-cross-reactive with antibodies against the porcine reproductive and respiratory syndrome virus, classical swine fever virus, porcine circovirus type 2, and pseudorabies virus. The maximum serum dilution for detection of ASFV-positive sera was 1:1600. The intra-batch reproducibility coefficient of variation was <5% and the inter-batch reproducibility coefficient of variation was <10%. Compared with commercial kits, the dual-antigen indirect ELISA had good detection performance. In conclusion, we established a detection method with low cost, streamlined production process, and fewer instruments. It provides a new method for the serological diagnosis of ASF.
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影响因子：
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