State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, Harbin 150069, China.
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, Harbin 150069, China.
J Virol Methods. 2020 May;279:113855. doi: 10.1016/j.jviromet.2020.113855. Epub 2020 Mar 12.
Porcine epidemic diarrhea virus (PEDV) causes very high mortality in newborn piglets. The mucosal immune system in the gut must eliminate potential pathogens while maintaining a mutually beneficial relationship with the commensal microbiota. Antibodies derived from the secretory immunoglobulin A (SIgA) class, act as the first line of antigen-specific immunity in the gut by recognizing both pathogens and commensals. Therefore, the measurement of SIgA levels is an important index in evaluating PEDV infections and immune status. A simple and rapid method for the detection of PEDV-specific SIgA using an immunochromatographic test strip has been developed; incorporating a colloidal gold-labeled anti-SIgA secretory component (SC) mAb probe for the detection of anti-PEDV-specific SIgA in swine. On the strip, a gold-labeled anti-SIgA SC mAb was applied to a conjugate pad; purified PEDV particles and goat anti-mouse antibodies were blotted onto a nitrocellulose membrane to form the test and control lines, respectively. Results showed that the immunochromatographic test strip had high sensitivity and specificity. When compared with enzyme-linked immunosorbent assay, kappa value suggesting that the strip could be used to detect PEDV specific SIgA in colostrum samples. Furthermore, the strip assay is rapid and easy to perform with no requirement for professional-level skills or equipment. We found that the immunochromatographic test strip was a rapid, sensitive, and reliable method for the identification of PEDV specific SIgA, indicating its suitability for epidemiological surveillance as well as vaccine immunity when studying PEDV.
猪流行性腹泻病毒（PEDV）可导致新生仔猪的高死亡率。肠道中的黏膜免疫系统必须在与共生菌群保持互利关系的同时，消除潜在的病原体。源自分泌型免疫球蛋白 A（SIgA）的抗体通过识别病原体和共生菌群，作为肠道中抗原特异性免疫的第一道防线。因此，SIgA 水平的测量是评估 PEDV 感染和免疫状态的重要指标。已经开发出一种使用免疫层析测试条检测 PEDV 特异性 SIgA 的简单快速方法；该方法将胶体金标记的抗 SIgA 分泌成分（SC）单克隆抗体探针用于检测猪中的抗 PEDV 特异性 SIgA。在测试条上，将金标记的抗 SIgA SC mAb 应用于结合垫；纯化的 PEDV 颗粒和山羊抗小鼠抗体被点样到硝酸纤维素膜上，分别形成测试线和控制线。结果表明，免疫层析测试条具有高灵敏度和特异性。与酶联免疫吸附测定相比，κ 值表明该测试条可用于检测初乳样本中的 PEDV 特异性 SIgA。此外，该条带检测方法快速且易于操作，无需专业技能或设备。我们发现，免疫层析测试条是一种快速、敏感、可靠的方法，用于鉴定 PEDV 特异性 SIgA，表明其适合于 PEDV 研究中的流行病学监测以及疫苗免疫。
