Enrofloxacin Rapid Detection in Aquatic Foods: Based on DNA Aptamer Sensor

Author:

Bao Xiuli12,Wang Siyuan12,Hao Qingfang12,Bai Yue12,Li Siying12,Zhang Shuai12,Zhang Lei12,Kang Xinxin12,Lyu Mingsheng12ORCID,Wang Shujun12

Affiliation:

1. Jiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, China

2. Co-Innovation Center of Jiangsu Marine Bio-Industry Technology, Jiangsu Ocean University, Lianyungang 222005, China

Abstract

Enrofloxacin (ENR) is widely used as a synthetic fluoroquinolone antibiotic for disease control in aquatic animals. ENR aptamers were screened in this study using the magnetic bead-SELEX method, and a graphene oxide fluorescent sensor was developed to detect the ENR residues in aquatic products. Firstly, ENR was conjugated to amino magnetic beads by amidation reaction, and then the aptamer sequences showing high affinity to ENR were screened step by step by using the SELEX screening method. Finally, after 10 rounds of SELEX screening, six candidate aptamers with high affinity were obtained. Among these, ENR-Apt 6 was selected based on its secondary structure features, high affinity (Kd = 35.08 nM), and high specificity to ENR. Furthermore, a fluorescent sensor was prepared using graphene oxide and ENR-Apt 6. The results showed that the linear range of the sensor could reach 600 nM (R2 = 0.986), while its optimal linear range was 1–400 nM (R2 = 0.991), with the lowest detection limit of 14.72 nM. The prepared sensor was successfully used for the detection of ENR in real samples, with a recovery range of 83.676–114.992% and a relative standard deviation < 10% for most of the samples.

Funder

Jiangsu Agricultural Science and Technology Innovation Fund

Priority Academic Program Development of Jiangsu Higher Education Institutions

Post-graduate Research and Practice Innovation Program of Jiangsu

Publisher

MDPI AG

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