Preliminary study on an innovative, simple mast cell-based electrochemical method for detecting foodborne pathogenic bacterial quorum signaling molecules (N-acyl-homoserine-lactones).
This paper reports the a novel and simple mast cell-based electrochemical method for detecting of bacterial quorum signaling molecules, N-acylhomoserine lactones (AHLs), which can be utilized to preliminarily evaluate the toxicity of food-borne pathogenic bacteria. Rat basophilic leukemia (RBL-2H3) mast cells encapsulated in alginate/graphene oxide hydrogel were immobilized on a gold electrode, while mast cells as recognition elements were cultured in a 3D cell culture system. Electrochemical impedance spectroscopy (EIS) was utilized to record the cell impedance signal as-influenced by Pseudomonas aeruginosa quorum-sensing molecule, N-3-oxododecanoyl homoserine lactone (3OC12-HSL). The results indicated that cellular activities such as cell viability, apoptosis, intracellular calcium, and degranulation were markedly influenced by the AHLs. Importantly, the exposure of 3OC12-HSL to mast cells induced a marked decrease in the electrochemical impedance signal in a dose-dependent manner. The detection limit for 3OC12-HSL was 0.034μM with a linear range of 0.1-1μM. These results were confirmed via conventional cell assay and transmission electron microscope (TEM) analysis. Altogether, the proposed method appears to be an innovative and effective approach to the quantitative measurement of Gram-negative bacterial quorum signaling molecules; to this effect, it also may serve as a primary evaluation of the cytotoxicity of food-borne pathogens.
School of Food Science and Technology, Yangzhou University, Yangzhou, Jiangsu 225127, PR China; jiangsu key labortary of zoonoses, Yangzhou, Jiangsu 225009, PR China.
Donglei
D
Jiang
School of Food Science and Technology, Yangzhou University, Yangzhou, Jiangsu 225127, PR China.
Dongdong
D
Feng
School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, PR China.
Hui
H
Jiang
Testing center, Yangzhou University, Yangzhou, Jiangsu 225009, PR China.
Limin
L
Yuan
School of Food Science and Technology, Yangzhou University, Yangzhou, Jiangsu 225127, PR China.
Yin
Y
Yongqi
School of Food Science and Technology, Yangzhou University, Yangzhou, Jiangsu 225127, PR China.
Xin
X
Xu
School of Food Science and Technology, Yangzhou University, Yangzhou, Jiangsu 225127, PR China. Electronic address: wmfang@yzu.edu.cn.
Weiming
W
Fang
D054742:Acyl-Butyrolactones / Q000737:chemistry / Q000302:isolation & purification*; D000818:Animals; D017209:Apoptosis / Q000235:genetics; D015374:Biosensing Techniques; D058266:Dielectric Spectroscopy; D005516:Food Microbiology; D008407:Mast Cells / Q000737:chemistry / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000302:isolation & purification* / Q000472:pathogenicity; D053038:Quorum Sensing / Q000235:genetics; D051381:Rats
D054742:Acyl-Butyrolactones
3OC(12)-HSL; EIS; Electrochemical assay; N-acyl-l-homoserine lactones; RBL-2H3 mast cell