A visual, rapid, and sensitive detection platform for Vibrio parahaemolyticus based on RPA-CRISPR/Cas12a and an immunochromatographic test strip
文献类型： 外文期刊
作者： Wang, Jinbin 1 ; Xu, Danhong 3 ; Liu, Hua 1 ; Liu, Juan 1 ; Zhu, Lemei 4 ; Zeng, Haijuan 1 ; Wu, Wenhui 3 ;
作者机构： 1.Shanghai Acad Agr Sci, Biotechnol Res Inst, Key Lab Agr Genet & Breeding, Shanghai, Peoples R China
2.Minist Agr & Rural Affairs, Crops Ecol Environm Secur Inspection & Supervis Ct, Key Lab Safety Assessment Agr Genet Modified Organ, Shanghai, Peoples R China
3.Shanghai Ocean Univ, Sch Food Sci & Technol, Shanghai, Peoples R China
4.Changsha Med Univ, Sch Publ Hlth, Academician Workstat, Changsha, Peoples R China
5.Shanghai Acad Agr Sci, Biotechnol Res Inst, 2901 Beidi Rd, Shanghai 201106, Peoples R China
6.Shanghai Ocean Univ, Sch Food Sci & Technol, 999 Huchenghuan Rd, Shanghai 201499, Peoples R China
关键词： CRISPR/Cas12a; immunochromatographic test strip; tlh; Vibrio parahaemolyticus
期刊名称：FOOD QUALITY AND SAFETY （ 影响因子：5.6； 五年影响因子：6.2 ）
ISSN： 2399-1399
年卷期： 2024 年 8 卷
页码：
收录情况： SCI
摘要： Objectives Vibrio parahaemolyticus is the primary species that causes vibriosis. In this study, a point-of-care detection method was developed for V. parahaemolyticus.Materials and Methods The detection platform targeted the thermolabile haemolysin (tlh) gene of V. parahaemolyticus based on recombinant polymerase amplification (RPA) and clustered regularly spaced short palindromic repeat (CRISPR/Cas) systems. The platform was combined with an immunochromatographic test strip (ICS) that enables low-cost, simple, visual detection of V. parahaemolyticus.Results The detection limit was 2.5x102 fg/mu L for plasmids and 1.4x102 CFU/mL for V. parahaemolyticus. In addition, V. parahaemolyticus in salmon sashimi could be detected at a concentration of 154 CFU/g without enrichment, and the entire detection time was around 30 min. After enrichment for 6 h, 2 CFU/g V. parahaemolyticus could be detected.Conclusions Consequently, the proposed RPA-CRISPR/Cas12a-ICS platform could detect V. parahaemolyticus in seafood intuitively, quickly, and sensitively, leading to high practical application value.
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