Development of a Fluorescently Labeled Aptamer Structure-Switching Assay for Sensitive and Rapid Detection of Gliotoxin.,Analytical Chemistry

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Development of a Fluorescently Labeled Aptamer Structure-Switching Assay for Sensitive and Rapid Detection of Gliotoxin.
Analytical Chemistry ( IF 6.7 ) Pub Date : 2018-12-20 00:00:00 , DOI: 10.1021/acs.analchem.8b05094
Shunxiang Gao _{1,

2,

3} , Xin Zheng ₄ , Yuan Tang ₅ , Yajun Cheng ₆ , Xiaobo Hu ₄ , Jihong Wu _{1,

2,

3}

Affiliation

Gliotoxin, one of the most toxic metabolites produced during the growth of Aspergillus fumigatus, can cause direct damage to the immune system and results in infection and spread of Aspergillus, or even leads to invasive aspergillosis. Accurate, rapid, and sensitive detection of the disease-specific marker gliotoxin, particularly in serum, urine, or other body fluids, is therefore an important approach to achieving early and rapid diagnosis of Invasive Aspergillus Fumigatus Infection (IAFI). In this study, aptamers that specifically bind to gliotoxin were successfully obtained using immobilization-free GO-SELEX technology. Furthermore, the performance of the aptamer, including binding affinity, targeting specificity, and structural stability, was further improved by optimizing through truncation and mutation. Finally, the optimized aptamer APT8T1M was used to develop a novel fluorescently labeled aptamer structure-switching assay (FLASSA) for the detection of gliotoxin. The method exhibited a good linear range from 0.1 nM to 100 nM of gliotoxin, with a lower detection limit of 0.05 nM. Moreover, FLASSA was applied to the detection of gliotoxin in spiked serum and urine samples. A good mean recovery of 98.76–110.85% and a low coefficient of variation (5.45–14.59%) were obtained, indicating a high degree of selectivity for gliotoxin, good reproducibility, and stability. These results show that the developed FLASSA has significant potential and offers an alternative to the traditional analytical methods for the rapid, sensitive, and efficient detection of gliotoxin, thus, providing an effective tool for the early and rapid diagnosis of IAFI.

中文翻译：

灵敏和快速检测胶质毒素的荧光标记适体结构转换分析的发展。

胶质毒素是烟曲霉生长过程中产生的最有毒的代谢产物之一，可对免疫系统造成直接损害，并导致曲霉菌的感染和扩散。，甚至导致侵袭性曲霉病。因此，准确，快速和灵敏地检测疾病特异性标记物gliotoxin，尤其是血清，尿液或其他体液中的胶质毒素，是实现早期和快速诊断烟熏曲霉感染（IAFI）的重要方法。在这项研究中，使用固定化的GO-SELEX技术成功获得了与胶质毒素特异性结合的适体。此外，通过截短和突变的优化，适体的性能，包括结合亲和力，靶向特异性和结构稳定性，得到了进一步改善。最后，使用优化的适体APT8T1M来开发新型荧光标记的适体结构转换测定法（FLASSA），用于检测神经胶质毒素。该方法显示出良好的线性范围（从0开始）。1 nM至100 nM的胶质毒素，检测下限为0.05 nM。此外，FLASSA还用于检测加标血清和尿液样品中的神经胶质毒素。获得了98.76–110.85％的良好平均回收率和较低的变异系数（5.45–14.59％），这表明对葡聚糖毒素的选择性高，再现性好，稳定性好。这些结果表明，开发的FLASSA具有巨大的潜力，并为传统，快速，灵敏和有效检测胶质毒素的分析方法提供了替代方法，从而为IAFI的早期和快速诊断提供了有效的工具。获得了59％），表明对葡聚糖毒素的选择性高，再现性好，稳定性好。这些结果表明，开发的FLASSA具有巨大的潜力，并为传统，快速，灵敏和有效检测胶质毒素的分析方法提供了替代方法，从而为IAFI的早期和快速诊断提供了有效的工具。获得了59％），表明对葡聚糖毒素的选择性高，重现性好，稳定性好。这些结果表明，开发的FLASSA具有巨大的潜力，并为传统，快速，灵敏和有效检测胶质毒素的分析方法提供了替代方法，从而为IAFI的早期和快速诊断提供了有效的工具。

更新日期：2018-12-20

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