猪德尔塔冠状病毒 S 基因截短片段的原核表达及间接 ELISA 检测方法的建立。
Prokaryotic expression of porcine deltacoronavirus S gene truncated segment and establishment of indirect ELISA detection method.
机构信息
College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China.
Key Laboratory of Animal Breeding and Genetics Key Laboratory of Sichuan Province, Sichuan Animal Science Academy, Chengdu, China; Livestock and Poultry Biological Products Key Laboratory of Sichuan Province, Sichuan Animal Science Academy, Chengdu, China.
出版信息
J Virol Methods. 2023 Oct;320:114775. doi: 10.1016/j.jviromet.2023.114775. Epub 2023 Jul 22.
Porcine deltacoronavirus (PDCoV) is an emerging discovered coronavirus that causes significant losses in the global swine industry. This study aimed to establish an indirect ELISA method for detecting PDCoV antibodies using the truncated gene of PDCoV spike protein (S). The purified S protein was used as the coating antigen for the polyclonal antibody. The conditions were optimized to establish an indirect ELISA detection method for PDCoV based on the S protein, which showed good specificity and no cross-reaction with SVV-VP1, ASFV-P72, GETV-E2, PRV-gE, etc. The method has high repeatability, with coefficients of variation within and between batches less than 10%. Compared with the commercial kit, the positive coincidence rate is 86.40%, the negative coincidence rate is 89.43%, and the total coincidence rate is 91.76%. This ELISA can be used for PDCoV serological investigation and antibody evaluation. It can also lay the foundation for further research and development of PDCoV S protein ELISA antibody detection kit.
猪德尔塔冠状病毒（PDCoV）是一种新兴的冠状病毒，它会给全球养猪业造成巨大损失。本研究旨在建立一种间接 ELISA 方法，用于检测 PDCoV 抗体，该方法使用 PDCoV 刺突蛋白（S）的截断基因。纯化的 S 蛋白被用作多克隆抗体的包被抗原。对基于 S 蛋白的 PDCoV 间接 ELISA 检测方法的条件进行了优化，该方法具有良好的特异性，与 SVV-VP1、ASFV-P72、GETV-E2、PRV-gE 等无交叉反应。该方法具有良好的重复性，批内和批间变异系数均小于 10%。与商品化试剂盒相比，该方法的阳性符合率为 86.40%，阴性符合率为 89.43%，总符合率为 91.76%。该 ELISA 可用于 PDCoV 的血清学调查和抗体评估。它也为进一步研究和开发 PDCoV S 蛋白 ELISA 抗体检测试剂盒奠定了基础。
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