Colorimetric Integrated PCR Protocol for Rapid Detection of Vibrio parahaemolyticus

• Main Authors: Kewen Cheng, Daodong Pan, Jun Teng, Li Yao, Yingwang Ye, Feng Xue, Fan Xia, Wei Chen
• Format: Article
• Language: English
• Published: MDPI AG 2016-09-01
• Series: Sensors
• Subjects: bacterium detection; colorimetric; PCR; primer design; virulence gene
• Online Access: http://www.mdpi.com/1424-8220/16/10/1600

Rapid detection of pathogens is of great significance for food safety and disease diagnosis. A new colorimetric method for rapid and easy detection of Vibrio parahaemolyticus (V. parahaemolyticus or 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 of tdh, trh, tlh and toxR virulence 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.