Fantoni Giulia, Deghmane Ala-Eddine, Caron François, Taha Muhamed-Kheir
Institut Pasteur, Invasive Bacterial Infections, Université Paris Cité, F-75006 Paris, France.
Department of Biotechnology, Chemistry and Pharmacy, University of Siena, 53100 Siena, Italy.
Microorganisms. 2025 Mar 4;13(3):595. doi: 10.3390/microorganisms13030595.
Serum bactericidal assay (SBA) is a functional assay that evaluates infection- and vaccine-induced neutralizing antibodies representing the serological correlate of protection against . However, it is time consuming due to its readout using the enumeration of colony-forming units (CFUs), making this conventional SBA (C-SBA) difficult for large-scale use. We developed a new SBA method that takes advantage of a bioluminescence serogroup B (BioLux-SBA). The assay development steps involved the human complement source validation, the setup of the optimal incubation time, and the assessment of intra-day and inter-day variability. BioLux-SBA was then compared to C-SBA using a serum collection of Norman children vaccinated in 2011 with MenBvac, an OMV meningococcal vaccine. While a conventional approach requests 48 h of work to test 24 sera per day, BioLux-SBA takes only 5 h to test 96 sera per day. The SBA titers (n = 10) correlated with R of 0.98 (-value < 0.0001). The deposition of terminal complement components (C5b-C9) measured by flow cytometry on the bacterial surface well correlated with BioLux SBA titers. This high-throughput method to evaluate the immunogenicity of meningococcal vaccines appears to be a reliable method for an OMV meningococcal B vaccine and requires further assessment in other laboratories and against other meningococcal vaccines.
血清杀菌试验（SBA）是一种功能性试验，用于评估感染诱导和疫苗诱导的中和抗体，这些抗体代表了针对……的保护性血清学关联指标。然而，由于其通过菌落形成单位（CFU）计数进行读数，耗时较长，使得这种传统的SBA（C-SBA）难以大规模使用。我们开发了一种利用生物发光的新型SBA方法——B群血清型生物发光SBA（BioLux-SBA）。试验开发步骤包括人补体来源验证、最佳孵育时间的设定以及日内和日间变异性评估。然后，使用2011年接种MenBvac（一种外膜囊泡脑膜炎球菌疫苗）的诺曼儿童的血清样本，将BioLux-SBA与C-SBA进行比较。传统方法每天检测24份血清需要48小时的工作量，而BioLux-SBA每天检测96份血清仅需5小时。SBA滴度（n = 10）的相关系数R为0.98（P值<0.0001）。通过流式细胞术测量的细菌表面终末补体成分（C5b-C9）沉积与BioLux SBA滴度高度相关。这种评估脑膜炎球菌疫苗免疫原性的高通量方法似乎是一种用于外膜囊泡B群脑膜炎球菌疫苗的可靠方法，需要在其他实验室针对其他脑膜炎球菌疫苗进行进一步评估。
