Shim Doo Hee, Kim Min Jung, Cha Hye-Ran, Park Eun Sun, Kim Ah Reum, Park Jeon Han, Park Hyung-Cheon, Song Daesub, Lee Jae Myun
Department of Microbiology and Immunology, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea.
Department of Pediatrics, Severance Hospital, Institute of Allergy, Severance Biomedical Science Institute, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea.
Clin Exp Vaccine Res. 2019 Jan;8(1):70-76. doi: 10.7774/cevr.2019.8.1.70. Epub 2019 Jan 31.
Enzyme-linked immunosorbent assay (ELISA) has been used in the diverse field to evaluate influenza virus infection; for the surveillance, diagnosis, efficacy evaluation, and development of the vaccine. The aim of this study was to establish an ELISA for detecting HA strain-specific antibodies using recombinant pandemic A H1N1 (pH1N1) HA1 (rHA1) protein.
rHA1 was produced in baculovirus system. The clinical performance of the developed ELISA was validated using human serum samples, by comparison with standard methods for detecting a neutralizing antibody; hemagglutination inhibition (HI) assay and microneutralization test (MNT). The ability of the ELISA system to evaluate the efficacy test of an influenza vaccine was explored by measuring antibody levels in the serum of vaccinated mice.
Our ELISA could detect anti-rHA1 antibody in influenza-infected patients and vaccinated subjects. Compared to HI assay and MNT as reference methods, our method showed good performance in detection of anti-rHA1 antibody. Detection of the anti-rHA1 antibody in vaccinated mice and its correlation with titers in HI assay was also proved in a mice model.
An ELISA system using rHA1 of pH1N1 influenza virus was developed, and showed good clinical performance in diagnosis of influenza virus infection and evaluation of the vaccination efficacy in both human and animal models.
酶联免疫吸附测定（ELISA）已在多个领域用于评估流感病毒感染，包括监测、诊断、疗效评估及疫苗研发。本研究的目的是建立一种使用重组甲型H1N1流感大流行病毒（pH1N1）血凝素1（HA1）蛋白（rHA1）检测HA株特异性抗体的ELISA方法。
rHA1在杆状病毒系统中产生。通过与检测中和抗体的标准方法（血凝抑制试验（HI）和微量中和试验（MNT））比较，使用人血清样本验证所建立ELISA的临床性能。通过检测接种疫苗小鼠血清中的抗体水平，探索ELISA系统评估流感疫苗效力试验的能力。
我们的ELISA能够检测流感感染患者和接种疫苗者体内的抗rHA1抗体。与作为参考方法的HI试验和MNT相比，我们的方法在检测抗rHA1抗体方面表现良好。在小鼠模型中也证实了接种疫苗小鼠体内抗rHA1抗体的检测及其与HI试验滴度的相关性。
开发了一种使用pH1N1流感病毒rHA1的ELISA系统，该系统在诊断流感病毒感染及评估人和动物模型中的疫苗接种效力方面均表现出良好的临床性能。
