Rapid, specific and sensitive detection of in seafood by accelerated strand exchange amplification

• Published in: Analytical methods Vol. 15; no. 5; pp. 655 - 662
• Main Authors: Sun, Ritong, Chen, Jiao, Wang, Yingeng, Zhang, Zheng, Li, Yong, Li, Fengmei, Ma, Cuiping, Han, Qingxia, Shi, Yanjing
• Format: Journal Article
• Published: 02.02.2023
• ISSN: 1759-9660; 1759-9679
• DOI: 10.1039/d2ay01889f

Vibrio parahaemolyticus infectious diseases caused by seafood contamination may be life-threatening to people with weak immunity. The detection of the Vibrio parahaemolyticus pathogen in aquatic foods is critical for reducing the outbreak of human Vibrio parahaemolyticus -associated diseases. In this study, a highly sensitive, specific, and time-saving real-time narrow thermal-cycling amplification detection method was developed based on accelerated strand exchange amplification (ASEA). It can detect cultured Vibrio parahaemolyticus at concentrations as low as 25 CFU mL −1 . In addition, for artificially spiked scallop meat, the detection limit was 1.8 × 10 3 CFU g −1 without pre-culture and 18 CFU g −1 of initial inoculum after 3 h enrichment. The whole assay, starting from DNA extraction, can be completed within 20 min. The ASEA detection method established in this study is an effective tool for the rapid detection of Vibrio parahaemolyticus strains in a large number of seafood samples. The total detection time from DNA preparation was about 20 min. The LOD of the ASEA method was 25 CFU mL −1 of pure cultured V. parahaemolyticus . The enrichment time was significantly reduced to 3 h with a LOD of 18 CFU g −1 of spiked scallop meat.