Berman Katherine, Van Slyke Greta, Novak Hayley, Rock Jean M, Bievenue Rachel, Damjanovic Amanda K, DeRosa Kate L, Mirabile Gianna, Girardin Roxie C, Dupuis Alan P, McDonough Kathleen A, Parker Monica M, Styer Linda M, Mantis Nicholas J
Division of Infectious Disease, Wadsworth Center, New York State Department of Health, Albany, NY 12208.
bioRxiv. 2024 Apr 2:2024.03.18.585599. doi: 10.1101/2024.03.18.585599.
In the earliest days of COVID-19 pandemic, the collection of dried blood spots (DBS) enabled public health laboratories to undertake population-scale seroprevalence studies to estimate rates of SARS-CoV-2 exposure. With SARS-CoV-2 seropositivity levels now estimated to exceed 94% in the United States, attention has turned to using DBS to assess functional (neutralizing) antibodies within cohorts of interest.
Contrived DBS eluates from convalescent, fully vaccinated and pre-COVID-19 serum samples were evaluated in SARS-CoV-2 plaque reduction neutralization titer (PRNT) assays, a SARS-CoV-2 specific 8-plex microsphere immunoassay, a cell-based pseudovirus assay, and two different spike-ACE2 inhibition assays, an in-house Luminex-based RBD-ACE2 inhibition assay and a commercial real-time PCR-based inhibition assay (NAB-Sure).
DBS eluates from convalescent individuals were compatible with the spike-ACE2 inhibition assays, but not cell-based pseudovirus assays or PRNT. However, the insensitivity of cell-based pseudovirus assays was overcome with DBS eluates from vaccinated individuals with high SARS-CoV-2 antibody titers.
SARS-CoV-2 neutralizing titers can be derived with confidence from DBS eluates, thereby opening the door to the use of these biospecimens for the analysis of vulnerable populations and normally hard to reach communities.
在新冠疫情最早期,干血斑(DBS)样本的采集使公共卫生实验室能够开展大规模人群血清流行率研究,以估计SARS-CoV-2的暴露率。目前估计美国SARS-CoV-2血清阳性率超过94%,人们的注意力已转向利用DBS来评估目标人群中的功能性(中和)抗体。
对康复期、完全接种疫苗以及新冠疫情前血清样本的人工合成DBS洗脱液进行了评估,采用了SARS-CoV-2空斑减少中和试验(PRNT)、一种SARS-CoV-2特异性8联微球免疫测定法、一种基于细胞的假病毒试验以及两种不同的刺突蛋白-血管紧张素转换酶2(spike-ACE2)抑制试验,即一种基于内部Luminex的RBD-ACE2抑制试验和一种基于商业实时PCR的抑制试验(NAB-Sure)。
康复期个体的DBS洗脱液与spike-ACE2抑制试验兼容,但与基于细胞的假病毒试验或PRNT不兼容。然而,高SARS-CoV-2抗体滴度的接种疫苗个体的DBS洗脱液克服了基于细胞的假病毒试验的不敏感性。
可以从DBS洗脱液中可靠地得出SARS-CoV-2中和滴度,从而为利用这些生物样本分析弱势群体和通常难以接触到的社区打开了大门。