Tu Yang-Chang, Chen Kuang-Yu, Chen Chung-Kung, Cheng Ming-Chu, Lee Shu-Hwae, Cheng Ivan-Chen
Department of Epidemiology, Animal Health Research Institute, Council of Agriculture, New Taipei City 25158, Taiwan.
Department of Veterinary Medicine, School of Veterinary Medicine, National Taiwan University, Taipei 10617, Taiwan.
J Vet Sci. 2019 Jan 31;20(1):51-57. doi: 10.4142/jvs.2019.20.1.51.
Monoclonal antibodies (MAbs) are widely applied in disease diagnoses. Herein, we report a MAb, WF-4, against nucleoprotein (NP), its broad response with , and its application in an immunohistochemistry (IHC) assay. WF-4 was screened by immunofluorescence assay (IFA). The results showed that its reactivity with baculovirus-expressed full-length recombinant NP (rNP) in Western blot (WB), indicating its IHC applicability. Fifteen (reference subtypes H1 to H15) infected chicken embryonated chorioallantoic membranes (CAM), fixed by formalin, were all detectable in the WF-4-based IHC assay. Also, the reactivity of the IHC test with NP from experimentally inoculated H6N1 and from all recent outbreaks of H5 subtype avian (AIV) field cases in Taiwan showed positive results. Our data indicate that CAM, a by-product of preparation, is helpful for -specific MAb characterization, and that the WF-4 MAb recognizes conserved and linear epitopes of NP. Therefore, WF-4 is capable of detecting NP antigens via IHC and may be suitable for developing various tests for diagnosis of and, especially, AIV infection.
单克隆抗体（MAbs）广泛应用于疾病诊断。在此，我们报告一种针对核蛋白（NP）的单克隆抗体WF-4，其广泛的反应性以及在免疫组织化学（IHC）检测中的应用。WF-4通过免疫荧光测定法（IFA）筛选。结果表明，其在蛋白质印迹法（WB）中与杆状病毒表达的全长重组NP（rNP）具有反应性，表明其适用于免疫组织化学检测。用福尔马林固定的15种甲型流感病毒（参考亚型H1至H15）感染的鸡胚绒毛尿囊膜（CAM），在基于WF-4的免疫组织化学检测中均能被检测到。此外，免疫组织化学检测对来自实验接种的H6N1以及台湾近期所有H5亚型禽流感病毒（AIV）田间病例暴发中分离的NP的反应性均呈阳性结果。我们的数据表明，CAM作为疫苗制备的副产品，有助于甲型流感病毒特异性单克隆抗体的鉴定，并且WF-4单克隆抗体识别甲型流感病毒NP的保守线性表位。因此，WF-4能够通过免疫组织化学检测NP抗原，可能适用于开发各种用于诊断甲型流感病毒尤其是禽流感病毒感染的检测方法。
