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纳米金核酸适配体HCR放大比色法检测 动物源食品中氨苄青霉素残留
张万方,何金娇,康瑞丽,朱艳平,马瑞,张同雨,王蕊,朱子任,郭晋汝,张晨雨,王选年
0
(新乡学院生命科学与基础医学学院,新乡 453003)
摘要:
为了检测动物原性食品中氨苄青霉素残留,利用氨苄青霉素与核酸适配体的特异性结合能使其脱离纳米金粒子表面的特性,并利用核酸适配体杂交链式反应(HCR)将信号放大,最终通过检测纳米金溶液吸收光谱的变化建立一种氨苄青霉素残留的高灵敏检测方法。首先对各项反应条件进行优化,筛选出HCR放大纳米金比色法的最优检测方法,优化后的检测方法中,NaCl溶液最优浓度为1 mol·L-1,HCR的最优终浓度为11.5% (V/V),HCR与纳米金孵育的最优反应条件为37 ℃、30 min。氨苄青霉素的检测范围是0~1.2 μmol·L-1,线性方程为y = 0.163 6 x + 0.025,R2 = 0.995 3,最低检测限可达到10 nmol·L-1,在实际样品中的检测回收率是92.30%~103.27%。建立的氨苄青霉素适配体的HCR放大纳米金比色法具有操作简便、反应快速、灵敏度高等优点,可用于氨苄青霉素残留量的快速检测。
关键词:  纳米金  核酸适配体  杂交链式反应  氨苄青霉素  比色法
DOI:10.13610/j.cnki.1672-352x.20210909.023
投稿时间:2020-11-19
基金项目:国家自然科学基金项目(81702074), 河南省科技攻关项目(182300410086)和河南省高等学校重点科研项目(20A180026)共同资助。
HCR amplification colorimetric assay for the detection of ampicillin residues in animal food based on gold nanoparticles
ZHANG Wanfang,HE Jinjiao,KANG Ruili,ZHU Yanping,MA Rui,ZHANG Tongyu,WANG Rui,ZHU Ziren,GUO Jinru,ZHANG Chenyu,WANG Xuannian
(School of Life Science & Basic Medicine, Xinxiang University, Xinxiang 453003)
Abstract:
In order to detect the residue of ampicillin in animal-derived food, a highly sensitive method for the detection of ampicillin residue was established. In this method, the aptamer recognized and binded to ampicillin with high specificity, leading to the aggregation of gold nanoparticles due to the loss of aptamer protection, and then the signal was amplified by HCR (hybridization chain reaction) and detected by the change of absorption spectra of nanometer gold solution. The results showed that the optimal concentration of NaCl solution and HCR was 1 mol·L-1 and 11.5% (V/V), respectively, and the optimal reaction conditions for HCR and nano-gold incubation was 37 ℃ for 30 min. Under this condition, the ampicillin detection range was from 0 to 1.2 μmol·L-1, the linear equation was y = 0.163 6 + 0.025 x, R2 = 0.995 3, and the lowest detection limit could be up to 10 nmol·L-1. The detection rates of the actual samples were from 92.30% to 103.27%. In conclusion, this paper established the ampicillin adapter HCR amplification of nanometer gold colorimetric method, which has the advantages of simple operation, quick response and high sensitivity, and can be used for the rapid detection of ampicillin residues.
Key words:  gold nanoparticles  aptamer  hybridization chain reaction (HCR)  ampicillin  colorimetric

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