Citation: ZHANG Qian-Wen, CHEN Qian, BIAN Xiao-Jun, YAN Juan. Aptasensor Based on Terminal Deoxynucleotidyl Transferase-Mediated Signal Amplification for Salmonella Typhimurium Detection[J]. Chinese Journal of Analytical Chemistry, ;2021, 49(8): 1289-1299. doi: 10.19756/j.issn.0253-3820.211148 shu

Aptasensor Based on Terminal Deoxynucleotidyl Transferase-Mediated Signal Amplification for Salmonella Typhimurium Detection

ZHANG Qian-Wen ¹ ,
CHEN Qian ¹ ,
BIAN Xiao-Jun ^1,2,3 ,
YAN Juan ^1,2,3,,

1.
College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China;
2.
Shanghai Engineering Research Center of Aquatic-Product Processing & Preservation, Shanghai 201306, China;
3.
Laboratory of Quality and Safety Risk Assessment for Aquatic Products on Storage and Preservation Shanghai, Ministry of Agriculture, Shanghai 201306, China

Corresponding author: YAN Juan, j-yan@shou.edu.cn
Received Date: 1 March 2021
Revised Date: 9 May 2021

Fund Project: Supported by the National Natural Science Foundation of China (No.21775102) and Natural Science Foundation of Shanghai Municipal, China (No.20ZR1424100).

Salmonella typhimurium (S.typhimurium) is one of the most widespread pathogens causing foodborne diseases. In this work, S.typhimurium was used as the model bacteria. An aptasensor based on aldehyde magnetic beads (Mbs), enzyme digestion and terminal deoxynucleotidyl transferase-mediated signal amplification technique for simple colorimetric detection of S.typhimurium was developed. The Mbs@dsDNA hybrid complex was first prepared using the capture probe (CP) and aptamer. In the presence of S.typhimurium, the aptamer specifically bound to it, and released from the Mbs, which was then removed by magnetic separation. Subsequently, CP on Mbs folded back on itself owing to partially complementary bases and formed enzyme digestion recognition sites. 3'-OH terminal ends were obtained by EcoR I enzyme digestion. Deoxynucleotides were catalyzed to the 3'-OH end under the action of terminal deoxynucleotidyl transferase (TdT) to obtain products of single-stranded DNA (ssDNA). In the process, signal reporter molecules were embedded in the chain of ssDNA, thus obtaining amplified signals. In the absence of S.typhimurium, TdT signal amplification reaction could not be initiated because no released aptamer and blocked 3'-OH end CP. The aptasensor had a good response performance to S.typhimurium in the concentration range of 10¹-10⁵ CFU/mL, and the detection limit was as low as 21 CFU/mL, showing high sensitivity and specificity. Moreover, the recovery of S.typhimurium in milk samples was 92.2%-112.7% (RSD<3%). The aptasensor showed potential application prospect in the food safety detection and supervision of other foodborne pathogenic microorganisms and other pollutants.
- Biosensor,
- Aptamer,
- Signal amplification,
- Foodborne diseases,
- Food safety
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