Detection of the Escherichia coli pathogenic gene eae with three real-time polymerase chain reaction methods.
Several real-time polymerase chain reaction (PCR) methods are currently available to rapidly detect the presence of a specific DNA sequence. When used for detection of pathogenic organisms, the turnaround time for PCR-based methods is much lower than for traditional culture techniques. This study compared the sensitivity of three real-time PCR methods when detecting the Escherichia coli pathogenic gene eae to determine which method is most effective in identifying very low levels of the organism. The three methods were used to detect the eae gene over a range of DNA concentrations. The differences in sensitivity were statistically significant (p<0.05), and SYBR Green I PCR was found to have the lowest detection limit of the three; LUX primers had the highest detection limit. Therefore, using a defined DNA concentration for detecting the eae gene, SYBR Green I is the best alternative.
McCrea
Joanne Karen
JK
National Microbiology Laboratory, Winnipeg, Manitoba, Canada.
Liu
Chenyi
C
Ng
Lai-King
LK
Wang
Gehua
G
D018829:Adhesins, Bacterial / Q000235:genetics*; D052160:Benzothiazoles; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D003959:Diamines; D019453:Escherichia coli O157 / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity*; D029968:Escherichia coli Proteins / Q000235:genetics*; D006801:Humans; D009930:Organic Chemicals; D016133:Polymerase Chain Reaction / Q000379:methods*; D011804:Quinolines; D012680:Sensitivity and Specificity; D019914:Taq Polymerase / Q000378:metabolism
D018829:Adhesins, Bacterial; D052160:Benzothiazoles; D017931:DNA Primers; D004269:DNA, Bacterial; D003959:Diamines; D029968:Escherichia coli Proteins; D009930:Organic Chemicals; D011804:Quinolines; C073819:eaeA protein, E coli; C098022:SYBR Green I; D019914:Taq Polymerase