Screening of broad-spectrum aptamer and development of electrochemical aptasensor for simultaneous detection of penicillin antibiotics in milk

Mengjiao Hu; Fengling Yue; Jiwei Dong; Chong Tao; Mengyuan Bai; Mengyue Liu; Shengxi Zhai; Shihao Chen; Wenzheng Liu; Guangyu Qi; Igor Vrublevsky; Xia Sun; Yemin Guo

doi:10.1016/j.talanta.2023.125508

Screening of broad-spectrum aptamer and development of electrochemical aptasensor for simultaneous detection of penicillin antibiotics in milk

Talanta. 2024 Mar 1:269:125508. doi: 10.1016/j.talanta.2023.125508. Epub 2023 Dec 5.

Authors

Affiliations

¹ College of Agricultural Engineering and Food Science, Shandong University of Technology, No. 266 Xincun Xilu, Zibo, Shandong 255049, China; Shandong Provincial Engineering Research Center of Vegetable Safety and Quality Traceability, No. 266 Xincun Xilu, Zibo, Shandong 255049, China; Zibo City Key Laboratory of Agricultural Product Safety Traceability, No. 266 Xincun Xilu, Zibo, Shandong 255049, China.
² Department of Information Security, Belarusian State University of Informatics and Radioelectronics, Minsk 220013, Belarus.
³ College of Agricultural Engineering and Food Science, Shandong University of Technology, No. 266 Xincun Xilu, Zibo, Shandong 255049, China; Shandong Provincial Engineering Research Center of Vegetable Safety and Quality Traceability, No. 266 Xincun Xilu, Zibo, Shandong 255049, China; Zibo City Key Laboratory of Agricultural Product Safety Traceability, No. 266 Xincun Xilu, Zibo, Shandong 255049, China. Electronic address: sunxia2151@sina.com.
⁴ College of Agricultural Engineering and Food Science, Shandong University of Technology, No. 266 Xincun Xilu, Zibo, Shandong 255049, China; Shandong Provincial Engineering Research Center of Vegetable Safety and Quality Traceability, No. 266 Xincun Xilu, Zibo, Shandong 255049, China; Zibo City Key Laboratory of Agricultural Product Safety Traceability, No. 266 Xincun Xilu, Zibo, Shandong 255049, China. Electronic address: gym@sdut.edu.cn.

PMID: 38070284
DOI: 10.1016/j.talanta.2023.125508

Abstract

Penicillin antibiotics (PENs) play an important role in killing pathogenic bacteria. However, the residues of various penicillin antibiotics in milk gradually accumulate in the human body with the increase of milk intake, which causes direct harm to the human body. Aptamers can be used as recognition element of sensors. It is great significance to use broad-spectrum aptamers for simultaneous detection of PENs. In this study, we reported the screening and identification of DNA aptamers for PENs. The aptamers were screened by graphene oxide-systematic evolution of ligands by exponential enrichment (GO-SELEX). The broad-spectrum aptamers with high affinity and specificity were successfully obtained after 13 rounds of screening. The affinity and specificity of candidate aptamers were analyzed by a GO fluorescence competition method. Further sequence analysis revealed that a truncated 47 nt aptamer (P-11-1) had a higher affinity than the original 79 nt aptamer. The truncated aptamer P-11-1 was used as a recognition element, and an electrochemical aptasensor was prepared using gold nanoparticles (AuNPs) combined with ferroferric oxide-multi walled carbon nanotube (Fe₃O₄-MWCNTs) complex. The results showed that the developed aptasensor achieved the simultaneous detection of PENs in milk samples across a concentration range of 2 nM-10,000 nM, achieving a limit of detection of 0.667 nM. This methodology provided a simple and sensitive new thinking for antibiotic multi-residue detection.

Keywords: Broad-spectrum aptamer; Electrochemical aptasensor; GO-SELEX; Milk samples; Penicillin antibiotics.

MeSH terms

Animals
Aptamers, Nucleotide* / chemistry
Biosensing Techniques* / methods
Gold / chemistry
Humans
Limit of Detection
Metal Nanoparticles* / chemistry
Milk / chemistry
Penicillins / analysis
SELEX Aptamer Technique / methods

Substances

Penicillins
Gold
P 11
Aptamers, Nucleotide