Center of Infectious Disease Control, National Institute of Public Health and the Environment (RIVM), Bilthoven, the Netherlands.
Virology Division, Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, the Netherlands.
mSphere. 2019 Apr 24;4(2):e00236-19. doi: 10.1128/mSphere.00236-19.
Human respiratory syncytial virus (RSV) is a major cause of severe respiratory disease in (premature) newborns and causes respiratory illness in the elderly. Different monoclonal antibody (MAb) and vaccine candidates are in development worldwide and will hopefully become available within the near future. To implement such RSV vaccines, adequate decisions about immunization schedules and the different target group(s) need to be made, for which the assessment of antibody levels against RSV is essential. To survey RSV antigen-specific antibody levels, we developed a serological multiplex immunoassay (MIA) that determines and distinguishes antibodies against the five RSV glycoproteins postfusion F, prefusion F, Ga, Gb, and N simultaneously. The standardized RSV pentaplex MIA is sensitive, highly reproducible, and specific for the five RSV proteins. The preservation of the conformational structure of the immunodominant site Ø of prefusion F after conjugation to the beads has been confirmed. Importantly, good correlation is obtained between the microneutralization test and the MIA for all five proteins, resulting in an arbitrarily chosen cutoff value of prefusion F antibody levels for seropositivity in the microneutralization assay. The wide dynamic range requiring only two serum sample dilutions makes the RSV-MIA a high-throughput assay very suitable for (large-scale) serosurveillance and vaccine clinical studies. In view of vaccine and monoclonal development to reduce hospitalization and death due to lower respiratory tract infection caused by RSV, assessment of antibody levels against RSV is essential. This newly developed multiplex immunoassay is able to measure antibody levels against five RSV proteins simultaneously. This can provide valuable insight into the dynamics of (maternal) antibody levels and RSV infection in infants and toddlers during the first few years of life, when primary RSV infection occurs.
人呼吸道合胞病毒（RSV）是导致早产儿严重呼吸道疾病的主要原因，也是老年人呼吸道疾病的病因。目前全球正在开发不同的单克隆抗体（MAb）和疫苗候选物，有望在不久的将来上市。为了实施这些 RSV 疫苗，需要对免疫接种计划和不同的目标人群做出充分的决策，而评估针对 RSV 的抗体水平是至关重要的。为了调查 RSV 抗原特异性抗体水平，我们开发了一种血清学多重免疫分析（MIA），可以同时检测和区分针对 RSV 融合后 F、融合前 F、Ga、Gb 和 N 这五种糖蛋白的抗体。这种标准化的 RSV 五聚体 MIA 具有较高的敏感性、重现性和特异性，可用于检测五种 RSV 蛋白。融合前 F 免疫显性区 Ø 与珠子偶联后结构构象的保留已得到证实。重要的是，MIA 与微量中和试验之间得到了很好的相关性，所有五种蛋白的结果均为阳性，这导致了在微量中和试验中选择一个任意的融合前 F 抗体水平截断值来作为血清阳性的标准。该 MIA 具有宽的动态范围，仅需两种血清稀释度，是一种高通量检测方法，非常适合（大规模）血清学监测和疫苗临床研究。鉴于疫苗和单克隆抗体的开发旨在降低因 RSV 引起的下呼吸道感染导致的住院和死亡，因此评估针对 RSV 的抗体水平至关重要。这种新开发的多重免疫分析能够同时测量针对五种 RSV 蛋白的抗体水平。这可以为婴儿和幼儿在生命的最初几年中（初次）RSV 感染期间，（母体）抗体水平和 RSV 感染的动态变化提供有价值的信息。
