Department of Chemistry, Vanderbilt University, Nashville, TN, 37615, USA.
Centers for Disease Control and Prevention, Atlanta, GA, 30341, USA.
Biosens Bioelectron. 2019 Apr 15;131:119-127. doi: 10.1016/j.bios.2019.01.056. Epub 2019 Jan 31.
Organophosphorus compounds (OPs) continue to represent a significant chemical threat to humans due to exposures from their use as weapons, their potential storage hazards, and from their continued use agriculturally. Existing methods for detection include ELISA and mass spectrometry. The new approach presented here provides an innovative first step toward a portable OP quantification method that surmounts conventional limitations involving sensitivity, selectivity, complexity, and portability. DNA affinity probes, or aptamers, represent an emerging technology that, when combined with a mix-and-read, free-solution assay (FSA) and a compensated interferometer (CI) can provide a novel alternative to existing OP nerve agent (OPNA) quantification methods. Here it is shown that FSA can be used to rapidly screen prospective aptamers in the biological matrix of interest, allowing the identification of a 'best-in-class' probe. It is also shown that combining aptamers with FSA-CI enables quantification of the OPNA metabolites, Sarin (NATO designation "G-series, B", or GB) and Venomous Agent X (VX) acids, rapidly with high selectivity at detection limits of sub-10 pg/mL in 25% serum (by volume in PBS). These results suggest there is potential to directly impact diagnostic specificity and sensitivity of emergency response testing methods by both simplifying sample preparation procedures and making a benchtop reader available for OPNA metabolite quantification.
有机磷化合物(OPs)由于其作为武器的使用、潜在的储存危害以及在农业上的持续使用,继续对人类构成重大的化学威胁。现有的检测方法包括 ELISA 和质谱法。这里提出的新方法为便携式 OP 定量方法提供了一个创新的第一步,克服了涉及灵敏度、选择性、复杂性和便携性的传统限制。DNA 亲和探针或适体代表了一种新兴技术,当与混合读取、无溶液检测(FSA)和补偿干涉仪(CI)结合使用时,可以为现有的 OP 神经毒剂(OPNA)定量方法提供一种新颖的替代方案。这里表明 FSA 可用于快速筛选感兴趣的生物基质中的潜在适体,从而鉴定出“最佳”探针。还表明,将适体与 FSA-CI 结合使用可以快速、高选择性地定量 OPNA 代谢物,沙林(北约命名为“G 系列,B”或 GB)和毒剂 X(VX)酸,在 25%血清中的检测限低至亚 10 pg/mL(PBS 中按体积计)。这些结果表明,通过简化样品制备程序并使台式读取器可用于 OPNA 代谢物定量,有可能直接影响应急响应测试方法的诊断特异性和灵敏度。