Colorimetric Integrated PCR Protocol for Rapid Detection of Vibrio parahaemolyticus.
Rapid detection of pathogens is of great significance for food safety and disease diagnosis. A new colorimetric method for rapid and easy detection ofVibrio parahaemolyticus(V. parahaemolyticusor Vp) has been developed in this research. A specific sequence was designed and integrated with the forward primer for molecular detection of Vp. This specific sequence was tested and treated as the horseradish peroxidase (HRP)-mimicking DNAzyme and could be amplified during the polymerase chain reaction (PCR) process. The products of PCR including the sequence of HRP-mimicking DNAzyme could produce the distinguished color in the presence of catalysis substrates. The optical signal of the catalysis reaction, which is in a linear relationship with the initial template of Vp, could be determined with the naked eye or measured with Ultraviolet-visible (UV-vis) for qualitative and quantitative detections, respectively. Based on the optical signal intensity, rapid and easy detection of Vp was successfully achieved with satisfied sensitivity and specificity. Furthermore, the detection oftdh,trh,tlhandtoxRvirulence genes of two Vp species (Vp 33847 and Vp 17802) were all performed successfully with this developed colorimetric integrated PCR protocol, which demonstrated potential applicability for the rapid detection of other bacteria.
School of Marine Science, Ningbo University, Ningbo 315211, China. chengkewen100@163.com.|School of Food Science & Engineering, Hefei University of Technology, Hefei 230009, China. chengkewen100@163.com.
Kewen
K
Cheng
School of Marine Science, Ningbo University, Ningbo 315211, China. daodongpan@163.com.
Daodong
D
Pan
School of Food Science & Engineering, Hefei University of Technology, Hefei 230009, China. guanpeng0189@163.com.
Jun
J
Teng
School of Food Science & Engineering, Hefei University of Technology, Hefei 230009, China. ybl200999@sohu.com.
Li
L
Yao
School of Food Science & Engineering, Hefei University of Technology, Hefei 230009, China. yeyingwang04@126.com.
Yingwang
Y
Ye
School of Food Science & Engineering, Hefei University of Technology, Hefei 230009, China. xuef1@jsciq.gov.cn.
Feng
F
Xue
School of Chemistry & Chemical Engineering, Huazhong University of Science & Technology, Wuhan 430074, China. xiafan@hust.edu.cn.
Fan
F
Xia
School of Food Science & Engineering, Hefei University of Technology, Hefei 230009, China. chenweishnu@hfut.edu.cn.
Wei
W
Chen
PCR; bacterium detection; colorimetric; primer design; virulence gene
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