H3N2 influenza hemagglutination inhibition method qualification with data driven statistical methods for human clinical trials.
H3N2 influenza hemagglutination inhibition method qualification with data driven statistical methods for human clinical trials.
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DOI：
10.3389/fimmu.2023.1155880
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发表时间：
2023
影响因子：
7.3
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中科院分区：
文献类型：
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季节性流感病毒株的血凝抑制（HAI）抗体滴度是疫苗诱导保护的重要替代物。然而，HAI检测在实验室之间可能是可变的，由于缺乏标准化，对不同病毒的灵敏度较低。在菌株特异性水平上对这些测定进行确认，能够精确和准确地定量HAI滴度。自1968年以来，甲型流感（H3 N2）仍然是欧洲和北美大多数国家的主要流行亚型，因此是流感疫苗研究的重点。作为美国国立卫生研究院（NIH）资助的合作流感疫苗创新中心（CIVICs）计划的一部分，我们报告了使用A/Texas/71/2017作为代表性H3 N2毒株和豚鼠红细胞和神经氨酸酶（NA）抑制剂奥司他韦（oseltamivir）来防止NA介导的凝集的HAI检测的稳健检测设计，严格的统计分析和完整的鉴定。该合格的HAI检测试剂盒对于中间精密度和操作员内变异性是精密的（通过几何变异系数（GCV）计算），通过相对误差计算是准确的，完全线性（斜率为-1，R-Square 1），稳健（<25%GCV），并具有高特异性和灵敏度。该HAI方法成功确认适用于另一种H3 N2流感病毒株A/新加坡/INFIMH-16-0019/2016，符合所有预先规定的验收标准。这些结果表明，新流感病毒株的HAI鉴定和数据生成可以有效地实现，只需最少的额外检测和开发。我们报告了一种合格的和适应性强的流感血清学方法和分析策略，用于测量可量化的HAI滴度，以确定人体临床试验中疫苗介导保护的相关性。
Hemagglutination inhibition (HAI) antibody titers to seasonal influenza strains are important surrogates for vaccine-elicited protection. However, HAI assays can be variable across labs, with low sensitivity across diverse viruses due to lack of standardization. Performing qualification of these assays on a strain specific level enables the precise and accurate quantification of HAI titers. Influenza A (H3N2) continues to be a predominant circulating subtype in most countries in Europe and North America since 1968 and is thus a focus of influenza vaccine research. As a part of the National Institutes of Health (NIH)-funded Collaborative Influenza Vaccine Innovation Centers (CIVICs) program, we report on the identification of a robust assay design, rigorous statistical analysis, and complete qualification of an HAI assay using A/Texas/71/2017 as a representative H3N2 strain and guinea pig red blood cells and neuraminidase (NA) inhibitor oseltamivir to prevent NA-mediated agglutination. This qualified HAI assay is precise (calculated by the geometric coefficient of variation (GCV)) for intermediate precision and intra-operator variability, accurate calculated by relative error, perfectly linear (slope of -1, R-Square 1), robust (<25% GCV) and depicts high specificity and sensitivity. This HAI method was successfully qualified for another H3N2 influenza strain A/Singapore/INFIMH-16-0019/2016, meeting all pre-specified acceptance criteria. These results demonstrate that HAI qualification and data generation for new influenza strains can be achieved efficiently with minimal extra testing and development. We report on a qualified and adaptable influenza serology method and analysis strategy to measure quantifiable HAI titers to define correlates of vaccine mediated protection in human clinical trials.