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首页> 外文期刊>Journal of Virological Methods >Development of a Coxsackievirus A16 neutralization assay based on pseudoviruses for measurement of neutralizing antibody titer in human serum
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Development of a Coxsackievirus A16 neutralization assay based on pseudoviruses for measurement of neutralizing antibody titer in human serum

机译：基于伪病毒的柯萨奇病毒A16中和测定方法的开发，用于测定人血清中的中和抗体效价

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摘要

Serum neutralizing antibody titers are indicative of protective immunity against Coxsackievirus A16 (CV-A16) and Enterovirus 71 (EV71), the two main etiological agents of hand, foot and mouth disease (HFMD), and provide the basis for evaluating vaccine efficacy. The current CV-A16 neutralization assay based on inhibition of cytopathic effects requires manual microscopic examination, which is time-consuming and labor-intensive. In this study, a high-throughput neutralization assay was developed by employing CV-A16 pseudoviruses expressing luciferase for detecting infectivity in rhabdomyosarcoma (RD) cells and measuring serum viral neutralizing antibodies. Without the need to use infectious CV-A16 strains, the neutralizing antibody titer against CV-A16 could be determined within 15 h by measuring luciferase signals by this assay. The pseudovirus CV-A16 neutralization assay (pCNA) was validated by comparison with a conventional CV-A16 neutralization assay (cCNA) in testing 174 human serum samples collected from children (age <5 years). The neutralizing antibody titers determined by these two assays were well correlated (R-2 = 0.7689). These results suggest that the pCNA can serve as a rapid and objective procedure for the measurement of neutralizing antibodies against CV-A16
机译：血清中和抗体滴度可指示针对手足口病（HFMD）的两种主要病原体-柯萨奇病毒A16（CV-A16）和肠病毒71（EV71）的保护性免疫力，并为评估疫苗效力提供基础。当前基于抑制细胞病变效应的CV​​-A16中和测定需要手动显微镜检查，这既费时又费力。在这项研究中，通过使用表达荧光素酶的CV-A16假病毒开发了高通量中和测定法，以检测横纹肌肉瘤（RD）细胞中的感染性并测量血清病毒中和抗体。无需使用感染性CV-A16菌株，可以通过此测定法测量荧光素酶信号，在15小时内确定针对CV-A16的中和抗体效价。伪病毒CV-A16中和测定法（pCNA）通过与常规CV-A16中和测定法（cCNA）进行比较，以测试从儿童（年龄小于5岁）收集的174个人血清样品。通过这两种测定确定的中和抗体滴度很好相关（R-2 = 0.7689）。这些结果表明，pCNA可以作为一种快速客观的方法来测量针对CV-A16的中和抗体

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