摘要
目的建立微囊藻毒素-LR(microcystin-LR, MC-LR)的纳米金生物传感比色检测法,并探究核酸适配体截短后对此方法检测效果的影响。方法采用柠檬酸钠还原法制备纳米金,通过优化盐浓度、适配体浓度、适配体与MC-LR的反应时间,建立了MC-LR的纳米金生物传感比色检测法。然后,采用组合型核酸适配体截短策略,探究了截短后的核酸适配体对比色法检测MC-LR的影响。结果 NaCl浓度为0.9mol/L,适配体浓度为0.4μmol/L, MC-LR与核酸适配体的反应时间为10 min时为其最适检测条件。在最适条件下,该方法的目测最低检测限为0.4μg/mL,仪器读数最低检测限为(1.05±0.002)ng/mL,线性检测范围为0~1.1μg/m L。探究得出核酸适配体截短后会影响其对MC-LR的识别活性。结论该方法简单、易行,线性检测范围宽,为小分子危害物核酸适配体的快速鉴定奠定了理论基础,同时,对截短核酸适配体与小分子危害物的识别活性研究提供了技术支撑。
Objective To establish a nano-gold biosensing colorimetric assay for microcystin-LR(MC-LR),and investigate the effect of nucleic acid aptamer truncation on the detection effect.Methods The gold nanoparticles were prepared by sodium citrate reduction method.After optimizing the concentration of salt,concentration of aptamer,reaction time,gold nanoparticles biosensor colorimetric method for the detection of MC-LR was developed.Then,the effect of nucleic acid aptamer contrast method on MC-LR detection was investigated by using combinatorial aptamer truncation strategy.Results The concentration of NaCl was 0.9 mol/L,the concentration of adaptor was 0.4 mumol/L,and the reaction time for MC-LR conjugation with aptamer was 10 min.Under the optimal conditions,the limit of detection(LOD)for MC-LR with naked eye was 0.4μg/mL,the LOD for instrument reading was(1.05±0.002)ng/mL,and the linear response range extended from 0 to 1.1μg/mL.It was concluded that nucleic acid aptamer truncation would affect its recognition activity to MC-LR.Conclusion This method is simple and easy to implement,and the linear detection range is wide.In addition,it lays a theoretical foundation for the rapid identification of aptamers specific for small molecules,and provides technical support for the recognition activity of truncate nucleic acid aptamers and small molecule hazards.
作者
陈思锐
侯建军
刘细霞
王芳
张婷
陆琪
CHEN Si-Rui;HOU Jian-Jun;LIU Xi-Xia;WANG Fang;ZHANG Ting;LU Qi(Hubei Key Laboratory of Edible Wild Plants Conservation and Utilization,Hubei Normal University,Huangshi,435002,China)
出处
《食品安全质量检测学报》
CAS
2019年第17期5748-5753,共6页
Journal of Food Safety and Quality
基金
国家自然科学基金项目(31601548)
湖北省自然科学基金项目(2016CFB218)
中央引导地方科技发展专项(2017ZYYD008)~~
