Detecting and genotyping Escherichia coli O157:H7 using multiplexed PCR and nucleic acid microarrays

Rapid detection and characterization of food borne pathogens such as Escherichia coli O157:H7 is crucial for epidemiological investigations and food safety surveillance. As an alternative to conventional technologies, we examined the sensitivity and specificity of nucleic acid microarrays for detect...

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Bibliographic Details
Published in:International journal of food microbiology 2001-07, Vol.67 (1), p.71-80
Main Authors: Call, Douglas R, Brockman, Fred J, Chandler, Darrell P
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Biotechnology
Chickens - microbiology
Colony Count, Microbial
DNA microarray
DNA microarrays
DNA, Bacterial - analysis
Electrophoresis, Agar Gel
Enterohemorrhagic
Escherichia coli
Escherichia coli O157 - classification
Escherichia coli O157 - genetics
Escherichia coli O157 - isolation & purification
Food industries
Food Microbiology
Food safety
Fundamental and applied biological sciences. Psychology
Gene Amplification
Genetic engineering
Genetic technics
Genotype
Hemolysin
In vitro gene amplification. Pcr technique
Intimin
Methods. Procedures. Technologies
Oligonucleotide Array Sequence Analysis - methods
Polymerase Chain Reaction - methods
Sensitivity and Specificity
Shiga-like toxin
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Summary:Rapid detection and characterization of food borne pathogens such as Escherichia coli O157:H7 is crucial for epidemiological investigations and food safety surveillance. As an alternative to conventional technologies, we examined the sensitivity and specificity of nucleic acid microarrays for detecting and genotyping E. coli O157:H7. The array was composed of oligonucleotide probes (25–30 mer) complementary to four virulence loci (intimin, Shiga-like toxins I and II, and hemolysin A). Target DNA was amplified from whole cells or from purified DNA via single or multiplexed polymerase chain reaction (PCR), and PCR products were hybridized to the array without further modification or purification. The array was 32-fold more sensitive than gel electrophoresis and capable of detecting amplification products from
ISSN:0168-1605
1879-3460
DOI:10.1016/S0168-1605(01)00437-8