Visual assay of Escherichia coli O157:H7 based on isothermal strand displacement and hybrid chain reaction amplification strategy

2021 ◽  
Author(s):  
Liu Xiu ◽  
Shengjun Bu ◽  
Hongguo Wei ◽  
Wang Ze ◽  
Songling Yu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
16S Rrna ◽  
Point Of Care ◽  
Escherichia Coli O157 ◽  
Strand Displacement ◽  
Chain Reaction ◽  
Reaction Amplification ◽  
Visual Assay ◽  
Amplification Strategy ◽  
Point Of Care Detection

Here, we describe a simple, sensitive, and enzyme-free method for visual point-of-care detection of 16S rRNA of Escherichia coli O157:H7 based on isothermal strand displacement-hybrid chain reaction (ISD-HCR) and lateral...

Polymerase Chain Reaction Assay for Detection of Escherichia coli O157:H7 and Escherichia coli O157:H−

2002 ◽  
Vol 65 (1) ◽  
pp. 5-11 ◽  
Author(s):  
TAKAHISA MIYAMOTO ◽  
NATSUKO ICHIOKA ◽  
CHIE SASAKI ◽  
HIROSHI KOBAYASHI ◽  
KEN-ICHI HONJOH ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Dna Sequence ◽  
Escherichia Coli O157 ◽  
Chain Reaction ◽  
Pcr Assay ◽  
E Coli ◽  
Pcr Product ◽  
Genomic Dnas ◽  
Polymerase Chain

The DNA band patterns generated by polymerase chain reaction (PCR) using the du2 primer and template DNAs from various strains of Escherichia coli and non–E. coli bacteria were compared. Among three to five prominent bands produced, the three bands at about 1.8, 2.7, and 5.0 kb were detected in all of the E. coli O157 strains tested. Some nonpathogenic E. coli and all pathogenic E. coli except E. coli O157 showed bands at 1.8 and 5.0 kb. It seems that the band at 2.7 kb is specific to E. coli O157. Sequence analysis of the 2.7-kb PCR product revealed the presence of a DNA sequence specific to E. coli O157:H− and E. coli O157:H7. Since the DNA sequence from base 15 to base 1008 of the PCR product seems to be specific to E. coli O157, a PCR assay was carried out with various bacterial genomic DNAs and O157-FHC1 and O157-FHC2 primers that amplified the region between base 23 and base 994 of the 2.7-kb PCR product. A single band at 970 bp was clearly detected in all of the strains of E. coli O157:H− and E. coli O157:H7 tested. However, no band was amplified from template DNAs from other bacteria, including both nonpathogenic and pathogenic E. coli except E. coli O157. All raw meats inoculated with E. coli O157:H7 at 3 × 100 to 3.5 × 102 CFU/25 g were positive both for our PCR assay after cultivation in mEC-N broth at 42°C for 18 h and for the conventional cultural method.

Rapid and accurate detection of Escherichia coli O157:H7 in beef using microfluidic wax-printed paper-based ELISA

The Analyst ◽  
2020 ◽  
Vol 145 (8) ◽  
pp. 3106-3115 ◽  
Author(s):  
Yanan Zhao ◽  
Dexin Zeng ◽  
Chao Yan ◽  
Wei Chen ◽  
Jianluan Ren ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Foodborne Pathogens ◽  
Point Of Care ◽  
Foodborne Pathogen ◽  
Escherichia Coli O157 ◽  
Good Sensitivity ◽  
Point Of Care Testing ◽  
Accurate Detection ◽  
New Ideas ◽  
Sensitivity Specificity

Escherichia coli O157:H7 is a severe foodborne pathogen. Paper-based ELISA can rapidly and accurately detect E.coli O157:H7 in beef. The method has good sensitivity, specificity and repeatability. It is suitable for point-of-care testing and offers new ideas for the detection of other foodborne pathogens.

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