A label-free and universal platform for antibiotics detection based on microchip electrophoresis using aptamer probes.
A novel label-free, universal, and high throughput aptasensor was developed based on a microchip electrophoresis (MCE) platform for automatic detection of antibiotic residues in food. Firstly, chloramphenicol (CAP) was employed as a model to be captured by its aptamer probe (Apt). Then, the partial complementary oligonucleotide of CAP's aptamer (C-DNA) was introduced into the reaction system. Because the Apt-CAP complex can't further hybrid with free C-DNA, the amount of hybrid Apt-C-DNA double strand DNA (dsDNA) was less than that without adding the target. Finally, the above mixture was introduced into the microchip electrophoresis (MCE) platform for detection, both dsDNA and Apt-CAP can be separated and produce different fluorescence signals in the MCE. In a certain concentration range, the ratio of signal between dsDNA and Apt-CAP (IdsDNA/IApt-CAP) was proportional to the concentration of targets. Under the optimum conditions, the ratio showed a satisfactory linearity range from 0.008 to 1ng/mL of CAP with a detection limit of 0.003ng/mL. Thus, a universal MCE-based assay was developed for quantifying CAP automatically. The method was also successfully applied in the different food samples for CAP detection, which showed a good recovery (Milk: 91.1-108%, Fish: 86.1-114%) and the results were consistent with that of ELISA. This method owned many merits as follows: firstly, MCE was a high throughput screening platform and the detection time is limited to 3min for each sample. Secondly, the aptamer probes can be directly used for detection without labeling any signal tag which can facilitate the preparation procedures of probes. Thirdly, the operation was easy just by the following steps: firstly, the mixture of aptamer probes were incubated followed adding C-DNA; then measurement was performed. Moreover, the assay with MCE platform can be used to detect other targets just by changing the corresponding aptamer probe; it can even realize simultaneous detection when the targets have aptamers with different number of base pairs. Above all, it's a high- throughput and prospective method which can be applied in high throughput screening of antibiotics in food safety.
Faculty of Material Science and Chemical Engineering, Ningbo University, Ningbo 315211, China.
Lingying
L
Zhou
Faculty of Material Science and Chemical Engineering, Ningbo University, Ningbo 315211, China. Electronic address: ganning@nbu.edu.cn.
Ning
N
Gan
Faculty of Material Science and Chemical Engineering, Ningbo University, Ningbo 315211, China.
You
Y
Zhou
Faculty of Material Science and Chemical Engineering, Ningbo University, Ningbo 315211, China.
Tianhua
T
Li
Faculty of Material Science and Chemical Engineering, Ningbo University, Ningbo 315211, China.
Yuting
Y
Cao
Department of Food Science and Engineering/Collaborative Innovation Center for Modern Grain Circulation and Safety, Nanjing University of Finance and Economics, Nanjing 210000, China.
Yinji
Y
Chen
D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis* / Q000737:chemistry; D052157:Aptamers, Nucleotide / Q000737:chemistry*; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D002417:Cattle; D002701:Chloramphenicol / Q000032:analysis* / Q000737:chemistry; D046209:Electrophoresis, Microchip / Q000295:instrumentation / Q000379:methods*; D005399:Fishes; D005504:Food Analysis / Q000295:instrumentation / Q000379:methods*; D005506:Food Contamination / Q000032:analysis*; D008892:Milk
D000900:Anti-Bacterial Agents; D052157:Aptamers, Nucleotide; D002701:Chloramphenicol
Chloramphenicol aptamer probe; Microchip electrophoresis; Microfluid; Screening Antibiotics