Microarray analysis of Escherichia coli O157:H7.

ABSTRACT:

Aim

To establish the rapid, specific, and sensitive method for detecting O157:H7 with DNA microchips.

Methods

Specific oligonucleotide probes (26-28 nt) of bacterial antigenic and virulent genes of E. coli O157:H7 and other related pathogen genes were pre-synthesized and immobilized on a solid support to make microchips. The four genes encoding O157 somatic antigen (rfbE), H7 flagellar antigen (fliC) and toxins (SLT1, SLT2) were monitored by multiplex PCR with four pairs of specific primers. Fluorescence-Cy3 labeled samples for hybridization were generated by PCR with Cy3-labeled single prime. Hybridization was performed for 60 min at 45 degrees. Microchip images were taken using a confocal fluorescent scanner.

Results

Twelve different bacterial strains were detected

SUBMITTER: Jin HY 

PROVIDER: S-EPMC4479681 | biostudies-literature | 2005 Oct

REPOSITORIES: biostudies-literature