基于杂交链式反应的适配体传感器用于卡那霉素的比色检测

引用本文: 田润, 陶晴, 卞晓军, 朱福琳, 颜娟. 基于杂交链式反应的适配体传感器用于卡那霉素的比色检测[J]. 分析化学, 2020, 48(5): 608-614. doi: 10.19756/j.issn.0253-3820.191708 shu

Citation: TIAN Run, TAO Qing, BIAN Xiao-Jun, ZHU Fu-Lin, YAN Juan. Aptasensor Based on Hybrid Chain Reaction for Colorimetric Detection of Kanamycin[J]. Chinese Journal of Analytical Chemistry, 2020, 48(5): 608-614. doi: 10.19756/j.issn.0253-3820.191708 shu

基于杂交链式反应的适配体传感器用于卡那霉素的比色检测

1.
上海海洋大学食品学院, 上海 201306;
2.
上海水产品加工及贮藏工程技术研究中心, 上海 201306;
3.
农业部水产品贮藏保鲜质量安全风险评估实验室(上海), 上海 201306
基金项目:
本文系国家自然科学基金项目（No.21775102）资助

摘要: 基于醛基化磁珠颗粒（Aldehyde magnetic beads，AMBs）和DNA杂交链式反应（Hybridization chain reaction，HCR）的信号放大策略，设计了检测卡那霉素（Kanamycin，Kana）的比色适配体生物传感器（Aptasensor）。制备了Mbs@DNA复合物，其上的卡那霉素适配体链与卡那霉素发生特异识别后，释放出互补链，互补链含有设计好的引发序列，可作为信号探针，引发HCR反应，生成的双链DNA（Double-stranded DNA，dsDNA）表现出强负电性，与溶液中带负电金纳米颗粒（Goldnanoparticles，AuNPs）相互排斥，在高盐条件下，AuNPs的稳定性被破坏而发生聚集，导致溶液的颜色从红色变为紫色，吸收光谱也发生变化。本方法检测卡那霉素的线性范围为1.6~32.0 nmol/L，检出限为0.9 nmol/L（S/N=3）；实际样品牛奶和蜂蜜样品中卡那霉素的加标回收率在96.0%~105.0%之间，相对标准偏差在3.3%~5.0%之间，说明本方法实用性良好。本方法对卡那霉素的检测具有较高的灵敏度和特异性，操作简单便捷，适用于现场检测等。同时，根据不同待测物具有与其特异性结合的适配体，本方法为其它目标物的检测提供了参考，在食品安全检测与监管中具有良好的应用前景。

关键词:

English

Aptasensor Based on Hybrid Chain Reaction for Colorimetric Detection of Kanamycin

1.
College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China;
2.
Shanghai Engineering Research Center of Aquatic-Product Processing & Preservation, Shanghai 201306, China;
3.
Laboratory of Quality and Safety Risk Assessment for Aquatic Products on Storage and Preservation (Shanghai), Ministry of Agriculture, Shanghai 201306, China
Received Date: 02 December 2019
Revised Date: 10 February 2020
Fund Project:
This work was supported by the National Natural Science Foundation of China (No. 21775102).

Abstract: The colorimetric aptasensors based on magnetic particle and hybridization chain reaction (HCR) for detection of kanamycin was developed. Kanamycin, an aminoglycoside antibiotic, is extensively used to treat bacterial infections by interrupting protein synthesis. However, the abuse and overuse of kanamycin may cause serious side effects, such as kidney damage, dizziness and hearing loss and so on. In this study, a kind of Mbs@ds-DNA complexes was prepared. The kanamycin aptamer on the Mbs@ds-DNA complexes could bind to kanamycin with strong specificity, while the complementary strand of kanamycin aptamer was used as a signal probe to initiate hybrid chain reaction (HCR). The product of HCR (dsDNA) provided less stabilization without ssDNA sticky ends, and the gold nanoparticles (AuNPs) would aggregate and exhibit a color change from red to purple under the condition of high salt, as well the UV/vis absorption spectra was also changed. The data showed that this method has high sensitivity and specificity for the detection of kanamycin. The linear range was 1.6-32.0 nmol/L with a detection limit of 0.9 nmol/L (S/N=3). Additionally, the proposed aptasensor was successfully utilized for detection of kanamycin in milk and honey samples, with recovery of 96.0%-105.0%, showing good practicability. Aptamer could specifically recognize and bind to target molecules, and therefore this assay provided the reference value for the detection of other targets, showing good potential in the aspects of food safety control and prevention.

Key words:

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1.
沈阳化工大学材料科学与工程学院沈阳 110142

基于杂交链式反应的适配体传感器用于卡那霉素的比色检测

English

Aptasensor Based on Hybrid Chain Reaction for Colorimetric Detection of Kanamycin

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