Increased Sensitivity of HIV-1 p24 ELISA Using a Photochemical Signal Amplification System.
Increased Sensitivity of HIV-1 p24 ELISA Using a Photochemical Signal Amplification System.
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DOI：
10.1097/qai.0000000000000726
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发表时间：
2015-10-01
期刊：
影响因子：
--
通讯作者：
Santockyte R
中科院分区：
文献类型：
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作者：
Bystryak S;Santockyte R
本文介绍了一种光化学信号放大法（PSAM），用于提高酶联免疫吸附试验（ELISA）检测HIV-1 p24抗原的灵敏度。该方法可用于市售和内部ELISA测试，并且具有比替代信号放大方法简单得多且成本更低的优点。光化学信号放大方法是基于辣根过氧化物酶（HRP）底物邻苯二胺（OPD）的自催化光化学反应。为了比较PSAM增强的ELISA与常规比色ELISA用于测定HIV-1 p24抗原的性能，我们采用PerkinElmer HIV-1 p24 ELISA试剂盒，使用常规ELISA和ELISA + PSAM。在本研究中，我们表明，PSAM技术允许一个增加的分析灵敏度和动态范围的商业HIV-1 p24 ELISA试剂盒，有和没有免疫复合物的破坏（ICD和非ICD ELISA），由约40倍的因素。ELISA + PSAM与市售微量滴定板读数器兼容，仅需要廉价的照明装置，并且PSAM扩增步骤不超过15分钟。
In this study we describe a photochemical signal amplification method (PSAM) for increasing of the sensitivity of enzyme-linked immunosorbent assay (ELISA) for determination of HIV-1 p24 antigen. This method can be used for both commercially available and in-house ELISA tests, and has the advantage of being considerably simpler and less costly than alternative signal amplification methods. The photochemical signal amplification method is based on an autocatalytic photochemical reaction of a horseradish peroxidase (HRP) substrate, orthophenylenediamine (OPD). To compare the performance of PSAM-boosted ELISA with a conventional colorimetric ELISA for determination of HIV-1 p24 antigen we employed a PerkinElmer HIV-1 p24 ELISA kit, using conventional ELISA alongside ELISA + PSAM. In the present study, we show that PSAM technology allows one to increase the analytical sensitivity and dynamic range of a commercial HIV-1 p24 ELISA kit, with and without immune-complex disruption (ICD and Non-ICD ELISA), by a factor of approximately 40-fold. ELISA + PSAM is compatible with commercially available microtiter plate readers, requires only an inexpensive illumination device, and the PSAM amplification step takes no longer than 15 min.