Sensitive Quantification of Escherichia coli O157:H7, Salmonella enterica, and Campylobacter jejuni by Combining Stopped Polymerase Chain Reaction with Chemiluminescence Flow-Through DNA Microarray Analysis

2011 ◽  
Vol 83 (8) ◽  
pp. 3153-3160 ◽  
Author(s):  
Simon Christian Donhauser ◽  
Reinhard Niessner ◽  
Michael Seidel
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Campylobacter Jejuni ◽  
Microarray Analysis ◽  
Dna Microarray ◽  
Salmonella Enterica ◽  
Escherichia Coli O157 ◽  
Chain Reaction ◽  
Flow Through ◽  
Polymerase Chain

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.

Evaluation of a Multiplex Real-Time Polymerase Chain Reaction for the Quantification of Escherichia coli O157 in Cattle Feces

2012 ◽  
Vol 9 (1) ◽  
pp. 79-85 ◽  
Author(s):  
Megan E. Jacob ◽  
Xiaorong Shi ◽  
Baoyan An ◽  
Tiruvoor G. Nagaraja ◽  
Jianfa Bai
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Real Time ◽  
Escherichia Coli O157 ◽  
Chain Reaction ◽  
Cattle Feces ◽  
Polymerase Chain
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