利用铕纳米粒子改进针对禽流感 H7 亚型病毒的单克隆抗体快速诊断应用。

Improvement of a rapid diagnostic application of monoclonal antibodies against avian influenza H7 subtype virus using Europium nanoparticles.

机构信息

Zoonosis Research Center, Department of Infection Biology, School of Medicine, Wonkwang University, 460, Iksan-daero, Iksan, 54538, Republic of Korea.

Department of Microbiology, Ajou University School of medicine, and Department of Biomedical Science, Graduate School of Ajou University, Suwon, Republic of Korea.

出版信息

Sci Rep. 2017 Aug 11;7(1):7933. doi: 10.1038/s41598-017-08328-9.

The development of a sensitive and rapid diagnostic test is needed for early detection of avian influenza (AI) H7 subtype. In this study, novel monoclonal antibodies (mAbs) against influenza A H7N9 recombinant hemagglutinin (rHA)1 were developed and applied to a Europium nanoparticle-based rapid fluorescent immunochromatographic strip test (FICT) to improve the sensitivity of the rapid diagnostic system. Two antibodies (2F4 and 6D7) exhibited H7 subtype specificity in a dot-FICT assay by optimization of the conjugate and the pH of the lysis buffer. The subtype specificity was confirmed by an immunofluorescence assay and Western blot analysis. The limit of detection of the FICT employing novel mAbs 31 ng/mL for H7N9 rHA1 and 40 hemagglutination units/mL for H7 subtype virus. Sensitivity was improved 25-fold using Europium as confirmed by comparison of colloidal gold-based rapid diagnostic kit using the 2F4 and 6D7 mAbs.

需要开发一种敏感且快速的诊断测试方法，以实现对禽流感（AI）H7 亚型的早期检测。在这项研究中，开发了针对甲型流感 H7N9 重组血凝素（rHA）1 的新型单克隆抗体（mAb），并将其应用于基于铕纳米颗粒的快速荧光免疫层析检测（FICT）中，以提高快速诊断系统的灵敏度。通过优化缀合物和裂解缓冲液的 pH 值，两种抗体（2F4 和 6D7）在斑点-FICT 检测中表现出 H7 亚型特异性。通过免疫荧光测定和 Western blot 分析证实了亚型特异性。新型 mAb 的 FICT 检测限为 31ng/mL 的 H7N9 rHA1 和 40 血凝单位/mL 的 H7 亚型病毒。通过比较使用 2F4 和 6D7 mAb 的胶体金为基础的快速诊断试剂盒，确认使用铕作为检测试剂可将灵敏度提高 25 倍。