Screening and development of DNA aptamers as capture probes for colorimetric detection of patulin

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• 作者：Shijia Wu ; Nuo Duan ; Weixiao Zhang ; Sen Zhao ; Zhouping Wang ; ^{wangzp1974@163.com" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Patulin ; Aptamer ; SELEX ; Graphene oxide ; Enzyme-chromogenic
• 刊名：Analytical Biochemistry
• 出版年：2016
• 出版时间：1 September 2016
• 年：2016
• 卷：508
• 期：Complete
• 页码：58-64
• 全文大小：898 K

文摘

Patulin (PAT) is a kind of mycotoxin that has serious harmful impacts on both food quality and human health. A high-affinity ssDNA aptamer that specifically binds to patulin was generated using systemic evolution of ligands by exponential enrichment (SELEX) assisted by graphene oxide (GO). After 15 rounds of positive and negative selection, a highly enriched ssDNA pool was sequenced and the representative sequences were subjected to binding assays to evaluate their affinity and specificity. Of the eight aptamer candidates tested, the sequence PAT-11 bound to patulin with high affinity and excellent selectivity with a dissociation constant (K_d) of 21.83 ± 5.022 nM. The selected aptamer, PAT-11, was subsequently used as a recognition element to develop a detection method for patulin based on an enzyme-chromogenic substrate system. The colorimetric aptasensor exhibited a linear range from 50 to 2500 pg mL⁻¹, and the limit of detection was found to be 48 pg mL⁻¹. The results indicated that GO-SELEX technology was appropriate for the screening of aptamers against small-molecule toxins, offering a promising application for aptamer-based biosensors.