Back to Search
Start Over
Microarray analysis of Escherichia coli O157:H7.
- Source :
-
World journal of gastroenterology [World J Gastroenterol] 2005 Oct 07; Vol. 11 (37), pp. 5811-5. - Publication Year :
- 2005
-
Abstract
- Aim: To establish the rapid, specific, and sensitive method for detecting O157:H7 with DNA microchips.<br />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.<br />Results: Twelve different bacterial strains were detected with various combinations of four virulent genes. All the O157:H7 strains yielded positive results by multiplex PCR. The size of the PCR products generated with these primers varied from 210 to 678 bp. All the rfbE/fliC/SLT1/SLT2 probes specifically recognized Cy3-labeled fluorescent samples from O157:H7 strains, or strains containing O157 and H7 genes. No cross hybridization of O157:H7 fluorescent samples occurred in other probes. Non-O157:H7 pathogens failed to yield any signal under comparable conditions. If the Cy3-labeled fluorescent product of O157 single PCR was diluted 50-fold, no signal was found in agarose gel electrophoresis, but a positive signal was found in microarray hybridization.<br />Conclusion: Microarray analysis of O157:H7 is a rapid, specific, and efficient method for identification and detection of bacterial pathogens.
- Subjects :
- Escherichia coli O157 pathogenicity
Escherichia coli Proteins analysis
Escherichia coli Proteins genetics
Fluorescent Dyes metabolism
Humans
Sensitivity and Specificity
Escherichia coli Infections diagnosis
Escherichia coli O157 genetics
Escherichia coli O157 isolation & purification
Molecular Diagnostic Techniques
Oligonucleotide Array Sequence Analysis
Subjects
Details
- Language :
- English
- ISSN :
- 1007-9327
- Volume :
- 11
- Issue :
- 37
- Database :
- MEDLINE
- Journal :
- World journal of gastroenterology
- Publication Type :
- Academic Journal
- Accession number :
- 16270390
- Full Text :
- https://doi.org/10.3748/wjg.v11.i37.5811