Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD, United States.
Chemical Sciences Division, Material Measurement Laboratory, National Institute of Standards and Technology, Gaithersburg, MD, United States.
Front Cell Infect Microbiol. 2021 Jan 11;10:620419. doi: 10.3389/fcimb.2020.620419. eCollection 2020.
Rapid, reliable, and sensitive detection of infection is central to malaria control and elimination. Many Malaria Rapid Diagnostic Tests (RDTs) developed for this purpose depend upon immunoassays that can be improved by advances in bound antibody sensor technology. In a previous study, immuno-polymerase chain reaction (PCR) was shown to provide highly sensitive detection of lactate dehydrogenase (PfLDH) in monoclonal antibody (mAb) sandwich assays. Here, we show comparably high immunoassay sensitivity by inductively coupled plasma mass spectrometry (ICP-MS) detection of gold nanoparticles (AuNPs). Following capture of PfLDH with the primary mAb and binding of the AuNP-labeled detection mAb, ICP-MS signals from the AuNPs provided quantitative measures of recombinant PfLDH test dilutions and infected erythrocytes. A detection limit of 1.5 pg/mL was achieved with the PfLDH protein. Parasitemia in cultures of -infected erythrocytes could be detected to a lower limit of 1.6 parasite/μl (p/μl) for early ring-stage forms and 0.3 p/μl for mixed stages including mature trophozoites and schizont-stages. These results show that ICP-MS detection of AuNPs can support highly sensitive and accurate detection of infection.
快速、可靠和敏感的感染检测是疟疾控制和消除的核心。为此目的开发的许多疟疾快速诊断检测(RDT)依赖于免疫测定法,而结合抗体传感器技术的进步可以改进这些免疫测定法。在之前的一项研究中,免疫聚合酶链反应(PCR)被证明在单克隆抗体(mAb)夹心测定中高度敏感地检测乳酸脱氢酶(PfLDH)。在这里,我们通过电感耦合等离子体质谱(ICP-MS)检测金纳米粒子(AuNP)显示出类似的高免疫测定灵敏度。在用初级 mAb 捕获 PfLDH 并结合 AuNP 标记的检测 mAb 后,AuNP 的 ICP-MS 信号提供了重组 PfLDH 测试稀释液和感染红细胞的定量测量。用 PfLDH 蛋白实现了 1.5 pg/mL 的检测限。-感染的红细胞培养物中的寄生虫血症可以检测到低至 1.6 个寄生虫/μl(p/μl)的早期环期形态和包括成熟滋养体和裂殖体阶段的混合阶段的 0.3 p/μl。这些结果表明,ICP-MS 检测 AuNP 可以支持对感染的高度敏感和准确检测。