A visual,rapid,and sensitive detection platform for Vibrio parahaemolyticus based on RPA-CRISPR/Cas12a and an immunochromatographic test strip

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外文摘要：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 2.5×102 fg/μL for plasmids and 1.4×102 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.

外文关键词：

CRISPR/Cas12aimmunochromatographic test striptlhVibrio parahaemolyticus

作者：

Jinbin Wang、Danhong Xu、Hua Liu、Juan Liu、Lemei Zhu、Haijuan Zeng、Wenhui Wu

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作者单位：

The Biotechnology Research Institute,Shanghai Academy of Agricultural Sciences,Key Laboratory of Agricultural Genetics and Breeding,Shanghai,China

Crops Ecological Environment Security Inspection and Supervision Center,Key Laboratory for Safety Assessment of Agricultural Genetically Modified Organisms,Ministry of Agriculture and Rural Affairs,Shanghai,China

School of Public Health,Academician Workstation,Changsha Medical University,Changsha,China

School of Food Science and Technology,Shanghai Ocean University,Shanghai,China

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出版年：

2024

DOI：

10.1093/fqsafe/fyae008

食品品质与安全研究(英文版)

ISSN：

年,卷(期)：2024.8(2)