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Two Laboratory Activities Using Conventional or Real-Time PCR to Simulate Pathogenic Detection

    Author: Joanna R. Klein1
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    Affiliations: 1: University of Northwestern – St. Paul, St. Paul, MN 55113
    AUTHOR AND ARTICLE INFORMATION AUTHOR AND ARTICLE INFORMATION
    • Published 01 May 2014
    • Supplemental materials available at http://jmbe.asm.org
    • Corresponding author. Mailing address: University of Northwestern – St. Paul, 3003 Snelling Ave. N., St. Paul, MN 55113. Phone: 651-286-7468. Fax: 651-286-7532. E-mail: jrklein@unwsp.edu.
    • ©2014 Author(s). Published by the American Society for Microbiology.
    Source: J. Microbiol. Biol. Educ. May 2014 vol. 15 no. 1 51-52. doi:10.1128/jmbe.v15i1.665
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    Abstract:

    In these lab activities, students perform conventional PCR and real-time PCR to simulate pathogenic detection. The labs were designed to complement a previously published virtual PCR classroom activity in which students are asked to design a PCR-based diagnostic test for a pathogenic strain of In the virtual PCR activity, students use bioinformatics to discover that the Shiga toxin genes ( and ) are unique to the Enterohemorrhagic (EHEC) strain O157:H7. Thus they come to the conclusion that doing PCR with primers designed for shiga toxin should be able to differentiate O157:H7 from other strains of In the lab activity described here, students actually perform the PCR assay. Performing PCR enhanced student understanding of the technique beyond what was accomplished through the virtual PCR classroom activity and is recommended as an addition to the case study.

Key Concept Ranking

Real-Time PCR
0.9374841
Shiga Toxins
0.5729167
0.9374841

References & Citations

1. BioRad 2006 Real-time PCR applications guide Bulletin 5279
2. Grys TE, Sloan LM, Rosenblatt JM, Patel R 2009 Rapid and sensitive detection of Shiga toxin-producing Escherichia coli from nonenriched stool specimens by real-time PCR in comparison to enzyme immunoassay and culture J. Clin. Microbiol 47 2008 2012 10.1128/JCM.02013-08 19439539 2708480 http://dx.doi.org/10.1128/JCM.02013-08
3. Klein JR, Gulsvig T 2012 Using bioinformatics to develop and test hypotheses: E. coli specific virulence determinants J. Microbiol. Biol. Educ 13 161 169 10.1128/jmbe.v13i2.451 http://dx.doi.org/10.1128/jmbe.v13i2.451
4. Madigan MT, Martinko JM, Dunlap PV, Clark DP 2009 Brock biology of microorganisms 12th ed Pearson Benjamin Cummings San Francisco, CA
5. Mellies JL, Barron AMS, Carmona AM 2007 Enteropathogenic and enterohemorrhagic Escherichia coli virulence gene regulation Infect. Immun 75 4199 4210 10.1128/IAI.01927-06 17576759 1951183 http://dx.doi.org/10.1128/IAI.01927-06
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/content/journal/jmbe/10.1128/jmbe.v15i1.665
2014-05-01
2017-09-20

Abstract:

In these lab activities, students perform conventional PCR and real-time PCR to simulate pathogenic detection. The labs were designed to complement a previously published virtual PCR classroom activity in which students are asked to design a PCR-based diagnostic test for a pathogenic strain of In the virtual PCR activity, students use bioinformatics to discover that the Shiga toxin genes ( and ) are unique to the Enterohemorrhagic (EHEC) strain O157:H7. Thus they come to the conclusion that doing PCR with primers designed for shiga toxin should be able to differentiate O157:H7 from other strains of In the lab activity described here, students actually perform the PCR assay. Performing PCR enhanced student understanding of the technique beyond what was accomplished through the virtual PCR classroom activity and is recommended as an addition to the case study.

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