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首页> 外文期刊>Applied Microbiology and Biotechnology >Optimization and validation of a blocking ELISA for quantitation of anti-rabies immunoglobulins in multispecies sera
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Optimization and validation of a blocking ELISA for quantitation of anti-rabies immunoglobulins in multispecies sera

机译：用于定量抗狂犬病免疫球蛋白血清血清的优化与验证

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摘要

We developed a fast, rabies virus-free, in vitro method, based on a blocking ELISA (bELISA), to detect and accurately quantify anti-rabies glycoprotein antibodies in serum of several animal species. In this method, purified rabies virus-like particles (VLPs) are used as antigen to coat the plates, while the presence of specific rabies immunoglobulins is revealed through blocking the recognition of these VLPs by a biotinylated monoclonal antibody. A quality by design approach was carried out in order to optimize the method performance, improving the sensitivity and, thereby, reducing the limit of detection of this assay. After the method validation, we confirmed that the bELISA method is able to detect a concentration of 0.06 IU/mL rabies immunoglobulins, titer lower than the 0.5 IU/mL cutoff value established as indication for correct vaccination. Further, we assessed the correlation between bELISA, the MNT, and the Platelia methods, confirming the accuracy of this new assay. On the other hand, precision was evaluated, obtaining acceptable repeatability and intermediate precision values, showing that this bELISA could be proposed as a potential alternative method, replacing the gold standard techniques in vaccination schemes and becoming a routine control technique within regional rabies surveillance programs.
机译：我们开发了一种快速的狂犬病病毒，体外方法，基于阻断ELISA（Belisa），以检测和准确地量化几种动物物种血清中的抗狂犬病糖蛋白抗体。在该方法中，纯化的狂犬病病毒样颗粒（VLP）用作抗原涂覆板，而通过阻断生物素化的单克隆抗体识别这些VLP的特异性狂犬病免疫球蛋白的存在。通过设计方法进行质量，以优化方法性能，提高灵敏度，从而降低该测定的检测限。在方法验证之后，我们证实Belisa方法能够检测0.06 IU / mL狂犬病免疫球蛋白的浓度，滴度低于0.5IU / mL截止值作为正确疫苗接种的指示。此外，我们评估了Belisa，MNT和血小板方法之间的相关性，确认了这种新测定的准确性。另一方面，评估精度，获得可接受的可重复性和中间精度值，表明该Belisa可以提出作为潜在的替代方法，取代疫苗接种方案中的金标准技术，成为区域狂犬病监测计划中的常规控制技术。

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