Calibration and Evaluation of Quantitative Antibody Titers for Measles Virus by Using the BioPlex 2200
Calibration and Evaluation of Quantitative Antibody Titers for Measles Virus by Using the BioPlex 2200
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DOI：
10.1128/cvi.00269-16
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发表时间：
2017-01-01
影响因子：
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通讯作者：
Severini, Alberto
中科院分区：
文献类型：
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作者：
Hatchette, Todd F.;Scholz, Heidi;Severini, Alberto
BioPlex 2200（Bio-Rad Laboratories，Hercules，CA）是一个快速、自动化的平台，可以筛选大量样本中的麻疹、腮腺炎、风疹和水痘抗体。尽管被批准用于产生定性结果，但在本研究中，我们验证了测试（标签外）以允许报告定量结果。为此，我们使用第三种抗麻疹世界卫生组织标准来生成校准曲线，使相对荧光强度能够转化为定量抗体滴度（抗体单位 [AU]/ml）。 BioPlex 2200 和参考噬菌斑减少中和试验 (PRNT) 的结果显示出遵循指数函数的合理相关性，但在低滴度样品中相关性较差。通过接受者操作特征分析，在 >= 0.13 和 < 1.10 AU/ml 之间建立了 BioPlex 2200 的模糊区域，以实现与 PRNT 相比的 100% 特异性（95% 置信区间 [CI] = 83.2 至 100%）和 100% 敏感性（95% CI = 93.5 至 100%）。通过确定需要 PRNT 确认的模糊范围，我们可以避免因假阴性结果低估免疫水平，并优化血清流行病学研究方法。
The BioPlex 2200 (Bio-Rad Laboratories, Hercules, CA) is a rapid, automated platform, which can screen large numbers of specimens for antibodies to measles, mumps, rubella, and varicella. Although approved for producing qualitative results, in this study we validated the test (off-label) to allow reporting of quantitative results. To do this, we used the third anti-measles World Health Organization standard to generate a calibration curve that allowed relative fluorescence intensity to be translated into quantitative antibody titer (antibody units [AU]/ml). The results from the BioPlex 2200 and the reference plaque reduction neutralization test (PRNT) exhibited a reasonable correlation following an exponential function, but correlation was poor in low-titer samples. Using a receiver operating characteristics analysis, an equivocal zone for the BioPlex 2200 was established between >= 0.13 and < 1.10 AU/ml to achieve 100% specificity (95% confidence interval [CI] = 83.2 to 100%) and 100% sensitivity (95% CI = 93.5 to 100%) versus PRNT. By determining an equivocal range requiring confirmation by PRNT, we can avoid underestimating the levels of immunity through false-negative results and optimize methods for seroepidemiological studies.