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Visual detection of Vibrio parahaemolyticus using gold nanoparticles and isothermal amplification

Visual detection of Vibrio parahaemolyticus using gold nanop...

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中国化学会第十三届全国分析化学年会

作者： Cong Kong Yuan Wang Guang-xin Yang Feng Han Dong-mei Huang Yin-feng Xi Run-run Gu Zhi-dong Liu Xiao-sheng Shen Key Laboratory of East China Sea Fishery Resources Exploitation Ministry of Agriculture East China Sea Fisheries Research Institute Chinese Academy of Fishery Sciences

Loop-mediated isothermal amplification（LAMP） eradicates the need of thermocycler in numerous fields related to molecular amplification detection and gives result through fluorescence or turbidity after the reaction, which is common used but still need to be improved for easier, simpler and more definitive readout. On the other side, Vibrio parahaemolyticus（V. parahaemolyticus） is one of the most prevalent seafood-borne pathogen worldwide, causing acute gastroenteritis. Here, a set of six specific primers targeting the species-specific tlh gene were used to realize the accelerated LAMP reaction. Gold nanoparticles（AuNPs） was functionalized with streptavidin（Avidin-AuNPs）, and engineered to signal the LAMP reaction. As two of the loop primers for LAMP reaction were biotinylated, the produced DNA product was able to mediate the clusterization of Avidin-AuNPs based on the avidin-biotin complex formation, which leads to the apparent color change from red to blue in the reaction solution. The rapid amplification could be performed within 30 min and readily identified by naked eye. The sensitivity, specificity and practicability of the nanosensing LAMP method were investigated and validated. This approach requires no fluorescent reagent and detection instrument. The Avidin-AuNPs can be used as simplified and generalized nanoprobes that are feasible for any similar LAMP product sensing. This simple, rapid, universal amplification and nanosensing strategy promised the high suitability of the developed method for point of care detection or utilization in resource-limited regions.

关键词： Loop-mediated isothermal amplification Gold Nanoparticles nanosense Vibrio parahaemolyticus

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