Research on nanogold-assisted HRM-qPCR technology for highly sensitive and accurate detection of Vibrio parahaemolyticus
Antuo Hu, Liangyu Kong, Zhaoxin Lu, Jiaju Qiao, Fengxia Lv, Fanqiang Meng, Xiaomei Bie
Abstract
The incidence of foodborne diseases ranks at the forefront of the total incidence of various diseases. High-specificity and high-sensitivity detection and reporting of foodborne pathogens have become the core of preventing and solving food safety problems. This study is based on screened highly specific molecular targets, the addition of the third-generation saturated fluorescent dyes and nanogold reduces the Ct value of the system while increasing the fluorescence signal value, thereby establishing highly sensitive and specific Nanogold-assisted HRM-qPCR rapid detection technology. The limit of detection and quantification of assay was 7 fg/μL-70 ng/μL for genomic DNA and 6 CFU/mL-6 × 106 CFU/mL for bacterial suspension. The Nanogold-assisted HRM-qPCR was shown to be able to detect Vibrio parahaemolyticus of 30 CFU/mL without enrichment culture and 3 CFU/mL after 2 h enrichment culture. This work shows promise in rapid instrumental and on-site detection of Vibrio parahaemolyticus.