|Title:||Rapid Detection of Food-Borne Escherichia coli O157:H7 with Visual Inspection by Crossing Priming Amplification (CPA)||Authors:||Xu Z
|Keywords:||Escherichia coli; Genes; Polymerase chain reaction; Toxic materials; Water pollution; Bacterial strains; Escherichia coli O157:H7; Food-borne pathogens; Limit of detection; Rapid detection; Reaction solutions; Shiga toxin; Visual inspection; Food microbiology||Issue Date:||2020||Journal Volume:||13||Journal Issue:||2||Start page/Pages:||474-481||Source:||Food Analytical Methods||Abstract:||
Escherichia coli O157:H7 is an important food-borne pathogen and has ability to contaminate food, such as water, meat products, and milk. Moreover, E. coli O157:H7 is the main toxin-producing serotype which can produce Shiga toxin type 1 and type 2 and cause intestinal disease. The strong pathogenicity and lethality of Escherichia coli O157:H7 pose a serious threat to human. This study aims to develop a rapid and visual detection assay of E. coli for rfbE, stx1, and stx2 genes by crossing priming amplification (CPA). The limit of detection of CPA assay for rfbE, stx1, and stx2 genes was 3.20 fg/μl, 320 fg/μl, and 320 fg/μl in genomic DNA, while that of in artificially contaminated food samples was 103 cfu/ml, 105 cfu/ml, and 105 cfu/ml, respectively, which was distinctly higher than that of PCR methods. And the specificity of CPA assay was tested by 22 different bacterial strains and except for E. coli O157:H7 ATCC43895 and other E. coli O157:H7 isolated from eggs, milk, and beef, all strains showed negative results. The visible detection assay was conducted by the addition of calcein in the reaction solutions. The CPA assay showed a successful detection of E. coli O157:H7 (Shiga toxin–producing and non-Shiga toxin–producing) within 60 min under 63 °C with high sensitivity and specificity. These results indicated that the CPA assays with calcein can provide an advanced method to achieve the rapid and visual detection of food-borne E. coli O157:H7. ? 2019, Springer Science+Business Media, LLC, part of Springer Nature.
Escherichia coli; Genes; Polymerase chain reaction; Toxic materials; Water pollution; Bacterial strains; Escherichia coli O157:H7; Food-borne pathogens; Limit of detection; Rapid detection; Reaction solutions; Shiga toxin; Visual inspection; Food microbiology
|Appears in Collections:||電機工程學系|
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.