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
Revised: April 25, 2002 |
Published: June 15, 2001
Citation
Call D.R., F.J. Brockman, and D.P. Chandler. 2001.Detecting and Genotyping Escherichia coli O157:H7 using multiplexed PCR and nucleic acid microarrays.International Journal of Food Microbiology 67, no. 1-2:71-80.PNNL-SA-34068.