Cheng Kewen, Pan Daodong, Teng Jun, Yao Li, Ye Yingwang, Xue Feng, Xia Fan, Chen Wei
School of Marine Science, Ningbo University, Ningbo 315211, China.
School of Food Science & Engineering, Hefei University of Technology, Hefei 230009, China.
Sensors (Basel). 2016 Sep 28;16(10):1600. doi: 10.3390/s16101600.
Rapid detection of pathogens is of great significance for food safety and disease diagnosis. A new colorimetric method for rapid and easy detection of ( 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 , , and 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.
病原体的快速检测对食品安全和疾病诊断具有重要意义。本研究开发了一种新的比色法，用于快速简便地检测（或Vp）。设计了一个特定序列，并将其与用于Vp分子检测的正向引物整合。该特定序列经过测试，并被视为辣根过氧化物酶（HRP）模拟DNAzyme，可在聚合酶链反应（PCR）过程中进行扩增。包含HRP模拟DNAzyme序列的PCR产物在催化底物存在下可产生明显的颜色。催化反应的光学信号与Vp的初始模板呈线性关系，可分别用肉眼或紫外可见（UV-vis）进行测定，以进行定性和定量检测。基于光学信号强度，成功实现了对Vp的快速简便检测，具有满意的灵敏度和特异性。此外，利用这种开发的比色整合PCR方案成功检测了两种Vp菌株（Vp 33847和Vp 17802）的、、和毒力基因，这表明该方法在快速检测其他细菌方面具有潜在的适用性。
