The Development of a Portable SPR Bioanalyzer for Sensitive Detection of Escherichia coli O157:H7.
The purpose of this study was to develop a portable surface plasmon resonance (SPR) bioanalyzer for the sensitive detection ofEscherichia coliO157:H7 in comparison with an enzyme-linked immunosorbent assay (ELISA). The experimental setup mainly consisted of an integrated biosensor and a homemade microfluidic cell with a three-way solenoid valve. In order to detectEscherichia coliO157:H7 using the SPR immunoassay, 3-mercaptopropionic acid (3-MPA) was chemisorbed onto a gold surface via covalent bond for the immobilization of biological species. 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) were used as crosslinker reagents to enable the reaction between 3-MPA andEscherichia coliO157:H7 antibodies by covalent -CO-NH- amide bonding. The experimental results were obtained from theEscherichia coliO157:H7 positive samples prepared by 10-, 20-, 40-, 80-, and 160-fold dilution respectively, which show that a good linear relationship with the correlation coefficient R of 0.982 existed between the response units from the portable SPR bioanalyzer and the concentration ofEscherichia coliO157:H7 positive samples. Moreover, the theoretical detection limit of 1.87 × 10³ cfu/mL was calculated from the positive control samples. Compared with theEscherichia coliO157:H7 ELISA kit, the sensitivity of this portable SPR bioanalyzer is four orders of magnitude higher than the ELISA kit. The results demonstrate that the portable SPR bioanalyzer could provide an alternative method for the quantitative and sensitive determination ofEscherichia coliO157:H7 in field.
Department of Electrical Engineering, Henan Agricultural University, Zhengzhou 45002, China. wangshun6518@163.com.|State Key Laboratory of Wheat and Maize Crop Science, Zhengzhou 45002, China. wangshun6518@163.com.
Shun
S
Wang
College of life sciences, Henan Agricultural University, Zhengzhou 45002, China. xiejiufeng0809@126.com.
Jiufeng
J
Xie
College of life sciences, Henan Agricultural University, Zhengzhou 45002, China. jm_hu@163.com.
Min
M
Jiang
Department of Electrical Engineering, Henan Agricultural University, Zhengzhou 45002, China. changkeke927@163.com.|State Key Laboratory of Wheat and Maize Crop Science, Zhengzhou 45002, China. changkeke927@163.com.
Keke
K
Chang
Department of Electrical Engineering, Henan Agricultural University, Zhengzhou 45002, China. chen_ruipeng@yeah.net.|State Key Laboratory of Wheat and Maize Crop Science, Zhengzhou 45002, China. chen_ruipeng@yeah.net.
Ruipeng
R
Chen
Department of Electrical Engineering, Henan Agricultural University, Zhengzhou 45002, China. mlz0124@126.com.|State Key Laboratory of Wheat and Maize Crop Science, Zhengzhou 45002, China. mlz0124@126.com.
Liuzheng
L
Ma
Department of Electrical Engineering, Henan Agricultural University, Zhengzhou 45002, China. zhujh88@sina.com.
Juanhua
J
Zhu
Department of Electrical Engineering, Henan Agricultural University, Zhengzhou 45002, China. guo_qingqian@163.com.|State Key Laboratory of Wheat and Maize Crop Science, Zhengzhou 45002, China. guo_qingqian@163.com.
Qingqian
Q
Guo
Department of Electrical Engineering, Henan Agricultural University, Zhengzhou 45002, China. haifengdreams@sina.cn.|State Key Laboratory of Wheat and Maize Crop Science, Zhengzhou 45002, China. haifengdreams@sina.cn.
Haifeng
H
Sun
Department of Electrical Engineering, Henan Agricultural University, Zhengzhou 45002, China. jdhu@henau.edu.cn.|State Key Laboratory of Wheat and Maize Crop Science, Zhengzhou 45002, China. jdhu@henau.edu.cn.
Jiandong
J
Hu
D015374:Biosensing Techniques / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D019453:Escherichia coli O157 / Q000502:physiology; D007118:Immunoassay / Q000379:methods; D013388:Succinimides; D020349:Surface Plasmon Resonance / Q000379:methods*
D013388:Succinimides; C001426:N-hydroxysuccinimide
ELISA kit; Escherichia coli O157:H7; biomolecular recognition membrane; portable surface plasmon resonance (SPR) bioanalyzer; response unit