Scalable, semi-automated fluorescence reduction neutralization assay for qualitative assessment of Ebola virus-neutralizing antibodies in human clinical samples

Antibody titers against a viral pathogen are typically measured using an antigen binding assay, such as an enzyme-linked immunosorbent assay (ELISA), which only measures the ability of antibodies to identify a viral antigen of interest. Neutralization assays measure the presence of virus-neutralizing antibodies in a sample. Traditional neutralization assays, such as the plaque reduction neutralization test (PRNT), are often difficult to use on a large scale due to being both labor and resource intensive. Here we describe an Ebola virus fluorescence reduction neutralization assay (FRNA), which tests for neutralizing antibodies, that requires only a small volume of sample in a 96-well format and is easy to automate. The readout of the FRNA is the percentage of Ebola virus-infected cells measured with an optical reader or overall chemiluminescence that can be generated by multiple reading platforms and the readout is compatible with lytic and non-lytic viruses. Using blinded human clinical...

针对病毒病原体的抗体滴度通常采用抗原结合试验测定，例如酶联免疫吸附试验（ELISA）——该方法仅能检测抗体识别目标病毒抗原的能力。中和试验用于检测样本中病毒中和抗体的存在。传统中和试验（如空斑减少中和试验，PRNT）通常因劳动和资源密集而难以大规模应用。我们在此描述一种埃博拉病毒荧光减少中和试验（FRNA）——该试验用于检测中和抗体，仅需少量样本，采用96孔板格式，且易于自动化。FRNA的读数结果为经光学读数仪测定的埃博拉病毒感染细胞百分比，或可通过多种读数平台生成的总化学发光值；该读数结果兼容裂解型和非裂解型病毒。使用盲法人类临床...