MURAL - Maynooth University Research Archive Library



    Real-time PCR methods for quantitative monitoring of streptomycin and tetracycline resistance genes in agricultural ecosystems


    Walsh, Fiona and Ingenfeld, A. and Zampicolli, M. and Hilber-Bodmer, M. and Frey, Jurg E. and Duffy, B. (2011) Real-time PCR methods for quantitative monitoring of streptomycin and tetracycline resistance genes in agricultural ecosystems. Journal of Microbiological Methods, 86 (2). pp. 150-155. ISSN 0167-7012

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    Abstract

    Antibiotic application in plant agriculture is primarily used to control fire blight caused by Erwinia amylovora in pome fruit orchards. In order to facilitate environmental impact assessment for antibiotic applications, we developed and validated culture-independent quantitative real-time PCR multiplex assays for streptomycin (strA, strB, aadA and insertion sequence IS1133) and tetracycline (tetB, tetM and tetW) resistance elements in plant and soil samples. The qPCR were reproducible and consistent whether the DNA was extracted directly from bacteria, plant and soil samples inoculated with bacteria or soil samples prior to and after manure slurry treatment. The genes most frequently identified in soils pre- and post-slurry treatment were strB, aadA, tetB and tetM. All genes tested were detected in soils pre-slurry treatment, and a decrease in relative concentrations of tetB and the streptomycin resistance genes was observed in samples taken post-slurry treatment. These multiplex qPCR assays offer a cost-effective, reliable method for simultaneous quantification of antibiotic resistance genes in complex, environmental sample matrices.

    Item Type: Article
    Keywords: Antibiotic resistance; Streptomycin; Tetracycline; Agriculture; qPCR; TaqMan®;
    Academic Unit: Faculty of Science and Engineering > Biology
    Item ID: 7475
    Identification Number: https://doi.org/10.1016/j.mimet.2011.04.011
    Depositing User: Fiona Walsh
    Date Deposited: 28 Sep 2016 15:30
    Journal or Publication Title: Journal of Microbiological Methods
    Publisher: Elsevier B.V.
    Refereed: Yes
    URI:

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