Detection of HIV-1 p24 antigen in patients with varying degrees of viremia using an ELISA with a photochemical signal amplification system.
Detection of HIV-1 p24 antigen in patients with varying degrees of viremia using an ELISA with a photochemical signal amplification system.
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DOI：
10.1016/j.cca.2016.02.022
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发表时间：
2016-05-01
期刊：
影响因子：
--
通讯作者：
Acharya C
中科院分区：
文献类型：
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作者：
Bystryak S;Acharya C
我们描述了一种光化学信号放大方法 (PSAM)，用于提高酶联免疫吸附测定 (ELISA) 检测 HIV-1 p24 抗原的灵敏度，并提出了 ELISA+PSAM 技术用于检测临床样本中 HIV-1 p24 抗原的初步验证研究。 ELISA+PSAM 与市售微量滴定板读数器兼容，采用廉价的照明设备，扩增大约需要 10 分钟。 PSAM技术不仅将HIV-1 p24抗原检测的分析灵敏度提高了约40倍，还显着提高了ELISA的临床灵敏度：在病毒RNA载量<3,000拷贝/ml的情况下，传统热介导免疫复合物破坏ELISA（HM-ELISA）无法检测到任何HIV阳性样本，而HM-ELISA+PSAM可以检测到约一半样本中的HIV感染（临床灵敏度为52.63%）。对于病毒RNA载量在3,000至30,000拷贝/ml之间，HM-ELISA和HM-ELISA+PSAM的临床敏感性分别为32.6%和91.3%，对于>30,000拷贝/ml，HM-ELISA和HM-ELISA+PSAM的临床敏感性分别为52.3%和100%。 HM-ELISA+PSAM 代表了全球医疗保健环境中监测 HIV-1 疾病进展和治疗的进步。
We describe a photochemical signal amplification method (PSAM) for increasing the sensitivity of enzyme-linked immunosorbent assays (ELISAs) for the detection of HIV-1 p24 antigen, and present a preliminary validation study on ELISA+PSAM technology for detection of HIV-1 p24 antigen in clinical samples. ELISA+PSAM is compatible with commercially available microtiter plate readers, employs an inexpensive illumination device and the amplification takes around 10 min. The PSAM technology not only increases the analytical sensitivity for detection of HIV-1 p24 antigen by approximately 40 times, but also significantly increases the clinical sensitivity of the ELISA: in instances where viral RNA load is <3,000 copies/ml, conventional heat mediated immune complex disruption ELISA (HM-ELISA) cannot detect any HIV positive samples whereas HM-ELISA+PSAM can detect HIV infection in approximately half of the samples (clinical sensitivity is 52.63%). For viral RNA loads between 3,000 and 30,000 copies/ml, the clinical sensitivities of the HM-ELISA and HM-ELISA+PSAM are 32.6% and 91.3%, and for that >30,000 copies/ml, clinical sensitivities of HM-ELISA and HM-ELISA+PSAM are 52.3% and 100%, respectively. The HM-ELISA+PSAM represents an advancement in monitoring HIV-1 disease progression and treatment in the global healthcare setting.