The diphtheria toxoid (DT), tetanus toxoid (TT), and acellular pertussis (aP) single immunogenicity assay (DTaP SIA) is a Luminex®-xMAP®-bead-based multiplex immunoassay for estimating the potency of DTaP pediatric combination vaccines in guinea pigs. This manuscript describes the validation of this assay for the simultaneous quantitation of anti-diphtheria toxoid (anti-DT), anti-tetanus toxoid (anti-TT), anti-pertussis toxoid (anti-PTxd), and anti-filamentous hemagglutinin (anti-FHA) antibodies in guinea pig serum following injection of a DTaP vaccine formulation. The results were expressed in arbitrary units/mL (AU/mL) using reference serum for comparison. Specificity was demonstrated by ≥ 75% homologous and ≤25% heterologous inhibition for all the antigens. The results were linear for anti-DT, anti-TT, anti-PTxd and anti-FHA antibodies. Accuracy was demonstrated with recovery of between 80% and 120% for all four antibodies. The relative standard deviation of repeatability was ≤20%. The results demonstrate that this SIA can be used for the linear, accurate, and precise simultaneous detection of all four antibodies, based on both the ICH Q2 and the EMA guidelines on bioanalytical method validation.

白喉类毒素（DT），破伤风类毒素（TT）和无细胞百日咳（aP）单免疫原性测定（DTaP SIA）是基于Luminex®-xMAP®-珠的多重免疫测定法，用于评估DTaP儿科联合疫苗在几内亚的效力猪。这个手稿描述了此测定法的抗白喉类毒素（的同时定量验证抗-DT），抗破伤风类毒素（抗-TT），抗百日咳类毒素（抗-PTxd），和抗丝状血凝素（抗注射DTaP疫苗制剂后，豚鼠血清中的-FHA）抗体。使用参考血清进行比较，结果以任意单位/ mL（AU / mL）表示。通过对所有抗原的≥75％同源和≤25％异源抑制证明了特异性。结果是线性为抗-DT，抗-TT，抗-PTxd和抗-FHA抗体。四种抗体的回收率均在80％至120％之间，证明了其准确性。重复性的相对标准偏差≤20％。结果表明，基于ICH Q2和EMA生物分析方法验证指南，该SIA可用于所有四种抗体的线性，准确和精确同时检测。