{"title":"Development of Recombinase Aided Amplification (RAA)-Exo-Probe Assay for the Rapid Detection of Shiga Toxin-Producing Escherichia coli.","authors":"Yuhao Cao, Taisong Fang, Jinling Shen, Guodong Zhang, Dehua Guo, Lina Zhao, Yuan Jiang, Shuai Zhi, Lin Zheng, Xiaofei Lv, Zhiyuan Yao, Daniel Yu","doi":"10.1093/jaoacint/qsad063","DOIUrl":null,"url":null,"abstract":"BACKGROUND\nShiga toxin-producing Escherichia coli (STEC) is a significant cause of foodborne illness causing various gastrointestinal diseases including hemolytic uremic syndrome (HUS), the most severe form, which can lead to kidney failure or even death.\n\n\nOBJECTIVE\nHere, we report the development of RAA (Recombinase Aided Amplification)-exo-probe assays targeting the stx1 and stx2 genes for the rapid detection of STEC in food samples.\n\n\nRESULTS\nThese assays were found to be 100% specific to STEC strains and were also highly sensitive with a detection limit of 1.6 × 103 CFU/mL or 32 copies/reaction. Importantly, the assays were able to successfully detect STEC in spiked and real food samples (beef, mutton, and pork), with a detection limit as low as 0.35 CFU/25g in beef samples after an overnight enrichment step.\n\n\nCONCLUSION\nOverall, the RAA assay reactions completed within ∼20 minutes and were less dependent on expensive equipment, suggesting they can be easily adopted for in field testing requiring only a fluorescent reader.\n\n\nHIGHLIGHTS\nAs such, we have developed two rapid, sensitive, and specific assays that can be used for the routine monitoring of STEC contamination in food samples, particularly in the field or in poorly equipped labs.","PeriodicalId":15003,"journal":{"name":"Journal of AOAC International","volume":"106 5","pages":"1246-1253"},"PeriodicalIF":1.7000,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of AOAC International","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1093/jaoacint/qsad063","RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}
引用次数: 0
Abstract
BACKGROUND
Shiga toxin-producing Escherichia coli (STEC) is a significant cause of foodborne illness causing various gastrointestinal diseases including hemolytic uremic syndrome (HUS), the most severe form, which can lead to kidney failure or even death.
OBJECTIVE
Here, we report the development of RAA (Recombinase Aided Amplification)-exo-probe assays targeting the stx1 and stx2 genes for the rapid detection of STEC in food samples.
RESULTS
These assays were found to be 100% specific to STEC strains and were also highly sensitive with a detection limit of 1.6 × 103 CFU/mL or 32 copies/reaction. Importantly, the assays were able to successfully detect STEC in spiked and real food samples (beef, mutton, and pork), with a detection limit as low as 0.35 CFU/25g in beef samples after an overnight enrichment step.
CONCLUSION
Overall, the RAA assay reactions completed within ∼20 minutes and were less dependent on expensive equipment, suggesting they can be easily adopted for in field testing requiring only a fluorescent reader.
HIGHLIGHTS
As such, we have developed two rapid, sensitive, and specific assays that can be used for the routine monitoring of STEC contamination in food samples, particularly in the field or in poorly equipped labs.
期刊介绍:
The Journal of AOAC INTERNATIONAL publishes the latest in basic and applied research in analytical sciences related to foods, drugs, agriculture, the environment, and more. The Journal is the method researchers'' forum for exchanging information and keeping informed of new technology and techniques pertinent to regulatory agencies and regulated industries.