INABIS '98 - Detection Of E.coli O157:H7 With A Quartz Crystal Biosensor And Asymmetric PCR

Detection Of E.coli O157:H7 With A Quartz Crystal Biosensor And Asymmetric PCR
Hurt, T. (Department of Chemistry and Biochemistry, University of Southern Mississippi, USA)
Craven, R.D. (Department of Chemistry and Biochemistry, University of Southern Mississippi, USA.)
Harvey, K.G. (Department of Chemistry and Biochemistry, University of Southern Mississippi, USA)
Zhang, P. (Department of Chemistry and Biochemistry, University of Southern Mississippi, USA)
Price, E. (Department of Chemistry and Biochemistry, University of Southern Mississippi, USA)
Fawcett, N.C. (Department of Chemistry and Biochemistry, University of Southern Mississippi, USA)
Evans, J.A. (Department of Chemistry and Biochemistry, University of Southern Mississippi, USA)

Contact Person: Jeffrey A. Evans (jeffrey.evans@usm.edu)

Abstract

It is estimated that Escherichia coli O157:H7 is responsible for more than 20,000 infections and as many as 250 deaths each year. Its early identification could prevent serious complications caused by this serotype. We are developing a rapid detection method for this pathogen by using a piezoelectric quartz crystal biosensor. The biosensor responds to an increase in mass on the crystal by a decrease in frequency. In our method, a DNA probe specific for the Shiga-like toxin (SLT) gene was attached to the crystal. We used asymmetric PCR to produce single-stranded target DNA by amplifying a region of the SLT-II gene from E. coli O157:H7. The amplified target was captured by the crystal bound probe. The quartz crystal responded to this mass increase by a decrease in its frequency, signaling that SLT-II DNA was present in the amplified sample. Asymmetric PCR eliminated the competing strand of DNA formed by regular PCR, increasing the target's ability to bind the crystal bound probe. Asymmetric PCR could generate single stranded targets of both the + and - strands. This procedure should be beneficial for detection of E. coli O157:H7.

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Poster Number PAhurt0758
Keywords: e coli O157:H7, asymmetric pcr, quartz crystal, biosensor, slt gene

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Hurt, T.; Craven, R.D.; Harvey, K.G.; Zhang, P.; Price, E.; Fawcett, N.C.; Evans, J.A.; (1998). Detection Of E.coli O157:H7 With A Quartz Crystal Biosensor And Asymmetric PCR. Presented at INABIS '98 - 5th Internet World Congress on Biomedical Sciences at McMaster University, Canada, Dec 7-16th. Available at URL http://www.mcmaster.ca/inabis98/newtech/hurt0758/index.html