Citation: HE Li-Yong, YU Jia-Le, LI Tian-Hua, CHEN Rong-Heng, LIN Jian-Yuan, GAN Ning. Simultaneous Detection of Vibrio Parahaemolyticus and Salmonella Typhimurium Based on Dual Mode Analysis Using Microfluidic Chip and Colorimetric Method[J]. Chinese Journal of Analytical Chemistry, ;2021, 49(1): 50-59. doi: 10.19756/j.issn.0253-3820.201354 shu

Simultaneous Detection of Vibrio Parahaemolyticus and Salmonella Typhimurium Based on Dual Mode Analysis Using Microfluidic Chip and Colorimetric Method

HE Li-Yong ^1,2 ,
YU Jia-Le ² ,
LI Tian-Hua ² ,
CHEN Rong-Heng ¹ ,
LIN Jian-Yuan ^1,, ,
GAN Ning ^2,,

1.
College of Biology and Environment, Zhejiang Wanli University, Ningbo 315100, China;
2.
Faculty of Material Science and Chemical Engineering, Ningbo Univeristy, Ningbo 315211, China

Corresponding author: LIN Jian-Yuan, GAN Ning,
Received Date: 18 June 2020
Revised Date: 22 November 2020

Fund Project: Supported by the National Natural Science Foundation of China (No.21974074).

Foodborne diseases caused by pathogenic microorganisms have attracted increasing attention in the world. It is urgent to develop efficient analytical methods for simultaneous and rapid detection of foodborne pathogens. In this regard, a dual-mode sensor based on the colorimetric principle coupled with microfluidic chip was developed using magnetic aptamer capture probes. It could realize rapid qualitative and simultaneous quantitative determination of Vibrio parahaemolyticus (V.P) and Salmonella typhimurium (S.T). The probe consisted several long DNA strands containing adenosine triphosphate (ATP), analyte aptamer and repeated G-base nucleotide fragments which were labeled on the surface of magnetic beads. When the probe reacted with the target analytes, the above long DNA strands would be released. The G quadruplet formed by hemin exhibited catalytic effect on tetramethylbenzidine (TMB)-H₂O₂ system, which could produce visible blue color and be used for on-site analysis of two pathogenic bacteria. After the long strand of released DNA was cut by EcoRV endonuclease, different lengths of DNA were obtained corresponding to V.P and S.T respectively. By analyzing each DNA in microchips, quantitative analysis of two kinds of bacteria V.P and S.T could be realized. Under the best experimental conditions, the detection linear range for the two bacteria was 10²-10⁷ CFU/mL, and the detection limit was 30 CFU/mL. The sensor provided a reliable method for screening contamination of pathogenic bacteria and quantification of each target accurately in the laboratory.
- Microfluidic chip,
- Colorimetric analysis,
- Rolling cycle amplification,
- Adenosine triphosphate,
- Vibrio parahaemolyticus,
- Salmonella typhimurium,
- Dual mode detection
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