Gao Fan, Xu Lingjie, Wang Qian, Wang Gang, Liu Mingchen, Li Lu, He Qian, Zhang Xuanxuan, Wang Ying, Mao Qunying, Liang Zhenglun, Wang Tao, Ma Xiao, Wu Xing
School of Life Sciences, Tianjin University, Tianjin 300072, China.
NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, NMPA Key Laboratory for Quality Research and Evaluation of Biological Products, State Key Laboratory of Drug Regulatory Science, Research Units of Innovative Vaccine Quality Evaluation and Standardization, Chinese Academy of Medical Sciences, National Institutes for Food and Drug Control, Beijing 102629, China.
Vaccines (Basel). 2025 Mar 18;13(3):320. doi: 10.3390/vaccines13030320.
BACKGROUND/OBJECTIVES: Enterovirus 71 (EV71) and coxsackieviruses A16 (CA16), A10 (CA10), and A6 (CA6) are the primary pathogens that cause hand, foot, and mouth disease (HFMD). Currently, many manufacturers are developing bivalent, trivalent, and tetravalent vaccines that target these antigens. Cell-based neutralization assay (CBNA), the gold standard for detecting neutralizing antibodies (NtAbs), which are used as indicators of HFMD vaccine efficacy, has several limitations. We aimed to develop a novel assay for detecting NtAbs against a quadrivalent HFMD vaccine.
We developed a four-color pseudovirus-based neutralization assay (PBNA), utilizing fluorescent reporter genes, to rapidly evaluate neutralizing antibodies against EV71, CA16, CA10, and CA6 in multivalent vaccines and compared it with CBNA.
PBNA could rapidly and simultaneously detect NtAbs against the four serotypes and required lesser amounts of sera compared to CBNA. A good consistency in determining NtAb titers was observed for PBNA and CBNA.
PBNA provides a robust tool for evaluating the efficacy of multivalent HFMD vaccines and conducting seroepidemiological studies.
背景/目的：肠道病毒71型（EV71）以及柯萨奇病毒A16型（CA16）、A10型（CA10）和A6型（CA6）是引起手足口病（HFMD）的主要病原体。目前，许多制造商正在研发针对这些抗原的二价、三价和四价疫苗。基于细胞的中和试验（CBNA）是检测中和抗体（NtAbs）的金标准，而中和抗体被用作手足口病疫苗效力的指标，但该试验存在一些局限性。我们旨在开发一种用于检测针对四价手足口病疫苗的中和抗体的新型试验。
我们利用荧光报告基因开发了一种基于四色假病毒的中和试验（PBNA），以快速评估多价疫苗中针对EV71、CA16、CA10和CA6的中和抗体，并将其与CBNA进行比较。
与CBNA相比，PBNA能够快速且同时检测针对这四种血清型的中和抗体，并且所需血清量更少。PBNA和CBNA在确定中和抗体滴度方面具有良好的一致性。
PBNA为评估多价手足口病疫苗的效力以及开展血清流行病学研究提供了一个强大的工具。
