Foodborne infectious diseases, particularly those caused by pathogenic Vibrio species in aquaculture, are a significant concern in the field of seafood safety. They not only hinder the economic development of the industry but also pose a major threat to public health. Traditional detection methods are often too slow, insensitive, or impractical for portable use, highlighting the urgent need for a sensitive, on-site detection method for Vibrio that prioritizes convenience and speed. This study developed a visual IMBS-CPCR method for the detection of four common pathogenic Vibrio species: Vibrio alginolyticus , Vibrio anguillarum , Vibrio fluvialis , and Vibrio harveyi . The method achieved high capture rates for target bacteria (82.6%–83.3%) and low capture rates for non-target bacteria (below 11%), demonstrating good sensitivity by detecting bacteria at concentrations as low as 10 CFU/mL. Optimal amplification conditions were determined for each species, with specificity tests confirming that only target bacteria produced amplification bands. Sensitivity tests revealed detection limits of 10 1 CFU/mL for V. alginolyticus , V. anguillarum , and V. fluvialis , and 10 2 CFU/mL for V. harveyi . The combined techniques improved detection sensitivity in simulated fish samples by 10–100 fold compared to direct CPCR or PCR methods.
Jiahui Li,Yuanyuan Zhang,Zhengshun Wen,Jiangbing Shuai,Xiaofeng Zhang,Jinru Zhou,Aiqian Ye,Linglin Fu
The delayed reporting of milk allergen residue detection results is a significant factor contributing to substantial losses for food production enterprises. Cumbersome operational procedures and high testing costs often lead to the abandonment of tests, resulting in frequent allergic reactions among milk-sensitive populations. Therefore, there is an urgent need for a novel, rapid, and low-cost detection method to mitigate enterprise losses and protect allergic individuals. This study investigates the formation mechanism of convection PCR, driven by Fast Taq polymerase in standard PCR tubes, and develops a fluorescence detection device for convection PCR. Through molecular docking and experimental validation of the trimmed β-LG aptamers, we elucidate their potential multi-site binding mechanism. Utilizing this mechanism, we designed a dual-strand hybridization magnetic bead detection system (dsA-SA-B), employing easily synthesized and amplified aptamer complementary strands as an indicator signal; Signal amplification was achieved within 15 min, while the entire detection process was completed in less than 50 min. This detection method allows for the rational selection of aptamer sequences based on actual testing needs, enabling the detection of hydrolyzed milk powder. The method demonstrated a linear detection range for β-LG from 5 to 100 μg/mL, with a detection limit as low as 0.25 μg/mL, effectively identifying hydrolyzed milk powder. This approach transforms protein detection into nucleic acid detection, and lays a solid foundation for multiplex PCR detection of eight major food allergens, facilitating the widespread detection of milk allergens and ultimately contributing to improved food safety and public health.
危险品化学品经营许可证（带存储）