State Key Laboratory of Food Science and Technology , Jiangnan University , Wuxi 214122 , China.
School of Food Science and Technology , Jiangnan University , Wuxi 214122 , China.
J Agric Food Chem. 2019 Feb 27;67(8):2313-2320. doi: 10.1021/acs.jafc.8b06893. Epub 2019 Feb 15.
In this work, after optimizing the original aptamer sequence by truncation and site-directed mutagenesis, a simple and sensitive colorimetric aptasensor was established for detecting the widespread food-borne pathogen Vibrio parahemolyticus ( V. parahemolyticus). The detection strategy was based on the competition for an V. parahemolyticus specific aptamer between its complementary DNA (cDNA) and V. parahemolyticus. The aptamer-conjugated magnetic nanoparticles (MNPs) were used as capture probes, and the G-quadruplex (G4) DNAzyme was employed as the signal amplifying element. Under optimal conditions, a wide linear detection range (from 10 to 10 cfu/mL) was available, and the detection limit could be as low as 10 cfu/mL. This method was also used to detect V. parahemolyticus in contaminated salmon samples, and the results showed good consistency with those obtained from standard plate counting method. Therefore, this novel aptasensor could be a good candidate for sensitive and selective detection of V. parahemolyticus without complicated operations.
在这项工作中，我们通过截短和定点突变对原始适体序列进行了优化，建立了一种简单、灵敏的比色适体传感器，用于检测广泛存在的食源性致病菌副溶血性弧菌（Vibrio parahemolyticus，V. parahemolyticus）。该检测策略基于副溶血性弧菌互补 DNA（cDNA）与副溶血性弧菌之间竞争特异性适体。将适体偶联的磁性纳米粒子（MNPs）用作捕获探针，G-四链体（G4）DNA 酶用作信号放大元件。在最佳条件下，该方法具有较宽的线性检测范围（从 10 到 10 cfu/mL），检测限可低至 10 cfu/mL。该方法还用于检测污染三文鱼样品中的副溶血性弧菌，结果与标准平板计数法的结果具有良好的一致性。因此，该新型适体传感器无需复杂操作即可用于副溶血性弧菌的灵敏选择性检测，是一种很有前途的方法。
