Development of Electrochemiluminescent Serology Assays to Measure the Humoral Response to Antigens of Respiratory Syncytial Virus

Sensitive and precise serology assays are needed to measure the humoral response to antigens of respiratory syncytial virus (RSV) following natural infection or vaccination. We developed and evaluated a collection of electrochemiluminescent (ECL) serology assays using four RSV antigens (F, N, Ga and Gb). To assess the merits of ECL technology, the four ECL serology assays were evaluated using a well-characterized “gold standard” panel of acute and convalescent serum samples from fifty-nine RSV-positive and thirty RSV-negative elderly subjects (≥65 years old). The combined results from the four ECL assays demonstrated good concordance to the “gold standard” diagnosis, reaching 95% diagnostic sensitivity and 100% diagnostic specificity. Additionally, a combination of ECL assays provided higher diagnostic sensitivity than a commercially available diagnostic ELISA or cell-based microneutralization assay. In summary, these data demonstrate the advantages of using ECL-based serology assays and highlight their use as a sensitive diagnostic approach to detect recent RSV infection in an elderly population.为检测自然感染或疫苗接种后机体针对呼吸道合胞病毒（respiratory syncytial virus, RSV）抗原的体液免疫应答，亟需灵敏且精准的血清学检测方法。本研究开发并评估了一套基于四种RSV抗原（F、N、Ga及Gb）的电化学发光（electrochemiluminescent, ECL）血清学检测方法。为评估ECL技术的应用价值，本研究利用一套经过充分验证的“金标准”样本库对这四种ECL血清学检测方法进行评估，该样本库包含59例RSV阳性及30例RSV阴性的老年受试者（年龄≥65岁）的急性期与恢复期血清样本。四种ECL检测方法的联合结果与“金标准”诊断具有良好一致性，诊断灵敏度达95%，诊断特异性达100%。此外，ECL检测联合方案的诊断灵敏度优于市售诊断用酶联免疫吸附试验（ELISA）及基于细胞的微量中和试验。总结而言，本研究数据证实了基于ECL的血清学检测方法的优势，并凸显了其作为灵敏诊断手段，用于检测老年人群近期RSV感染的应用价值。